Molecular evolution of coxsackievirus A24v in Cuba over 23-years, 1986-2009.

Coxsackievirus A24 variant (CVA24v) is a major causative agent of acute hemorrhagic conjunctivitis outbreaks worldwide, yet the evolutionary and transmission dynamics of the virus remain unclear. To address this, we analyzed and compared the 3C and partial VP1 gene regions of CVA24v isolates obtained from five outbreaks in Cuba between 1986 and 2009 and strains isolated worldwide. Here we show that Cuban strains were homologous to those isolated in Africa, the Americas and Asia during the same time period. Two genotypes of CVA24v (GIII and GIV) were repeatedly introduced into Cuba and they arose about two years before the epidemic was detected. The two genotypes co-evolved with a population size that is stable over time. However, nucleotide substitution rates peaked during pandemics with 4.39 × 10-3 and 5.80 × 10-3 substitutions per site per year for the 3C and VP1 region, respectively. The phylogeographic analysis identified 25 and 19 viral transmission routes based on 3C and VP1 regions, respectively. Pandemic viruses usually originated in Asia, and both China and Brazil were the major hub for the global dispersal of the virus. Together, these data provide novel insight into the epidemiological dynamics of this virus and possibly other pandemic viruses.

Transmissibility of acute haemorrhagic conjunctivitis in small-scale outbreaks in Hunan Province, China.

Acute haemorrhagic conjunctivitis (AHC) outbreaks are reported frequently in China. However, the transmissibility of AHC remains unclear. This study aimed to calculate the transmissibility of the disease with and without interventions. An AHC outbreak dataset from January 2007 to December 2016 in different schools was built in Hunan Province. A Susceptible-Infectious-Recovered (SIR) model was adopted to calculate the effective reproduction number (Reff) of AHC. Reff was divided into two parts (Runc and Rcon) where Runc and Rcon represent the uncontrolled and controlled Reff , respectively. Based on Runc and Rcon, an index of effectiveness of countermeasures (Ieff) was developed to assess the effectiveness of countermeasures in each outbreak. During the study period, 34 AHC outbreaks were reported in 20 counties of 9 cities in Hunan Province, with a mean total attack rate of 7.04% (95% CI: 4.97-9.11%). The mean Runc of AHC outbreaks was 8.28 (95% CI: 6.46-10.11). No significance of Runc was observed between rural and urban areas (t = -1.296, P = 0.205), among college, secondary, and primary schools (F = 0.890, P = 0.459), different levels of school population (F = 0.738, P = 0.538), and different number of index cases (F = 1.749, P = 0.180). The most commonly implemented countermeasures were case isolation, treatment, and health education, followed by environment disinfection, symptom surveillance, and school closure. Social distance, prophylaxis, and stopping eye exercises temporary were implemented occasionally. The mean value of Rcon was 0.16 (range: 0.00-1.50). The mean value of Ieff was 97.16% (range: 71.44-100.00%). The transmissibility of AHC is high in small-scale outbreaks in China. Case isolation, treatment, and health education are the common countermeasures for controlling the disease.

A modeling and experiment framework for the emergency management in AHC transmission.

Emergency management is crucial to finding effective ways to minimize or even eliminate the damage of emergent events, but there still exists no quantified method to study the events by computation. Statistical algorithms, such as susceptible-infected-recovered (SIR) models on epidemic transmission, ignore many details, thus always influencing the spread of emergent events. In this paper, we first propose an agent-based modeling and experiment framework to model the real world with the emergent events. The model of the real world is called artificial society, which is composed of agent model, agent activity model, and environment model, and it employs finite state automata (FSA) as its modeling paradigm. An artificial campus, on which a series of experiments are done to analyze the key factors of the acute hemorrhagic conjunctivitis (AHC) transmission, is then constructed to illustrate how our method works on the emergency management. Intervention measures and optional configurations (such as the isolation period) of them for the emergency management are also given through the evaluations in these experiments.

Isolation of Coxsackievirus A24 variant from patients with hemorrhagic conjunctivitis in Cuba, 2008-2009.

BACKGROUND: An outbreak of acute hemorrhagic conjunctivitis occurred in Cuba in 2008 and 2009. OBJECTIVE: To determinate the etiological agent associated with the Cuban outbreaks of acute hemorrhagic conjunctivitis during 2008 and 2009. STUDY DESIGN: Conjunctival swabs and/or faecal samples from 382 patients with clinical diagnosis suggestive of acute hemorrhagic conjunctivitis were subject to viral culture in HEp-2 human laryngeal epidermoid carcinoma cells. Positive samples were identified by a specific Coxsackievirus A24 variant PCR and the 3C protease region of 16 isolates was sequenced for phylogenetic analysis. RESULTS: Enterovirus cytopathic effect was observed in 138 cases (36%). A higher percent of CA24v was recovered from faecal samples, 19 out of 45 cases (42.2%), than from conjunctival swabs, 127 out of 355 samples (35.8%). All isolates were identified as Coxsackievirus A24 variant. Phylogenetic analysis revealed that 2008 and 2009 Cuban outbreaks were caused by the same virus strains and that isolates were closely related to those from Taiwan (2006-2007), China (2007-2008) and Singapore (2005) with a bootstrap value of 71%. CONCLUSIONS: Outbreaks of acute hemorrhagic conjunctivitis occurred in Cuba in 2008 and 2009 were caused by Coxsackievirus A24 variant. The faecal-oral route is another mode of transmission of CA24v in the acute hemorrhagic conjunctivitis outbreaks. Phylogenetic analysis of Cuban CA24v strains involved in an acute hemorrhagic conjunctivitis outbreak in 2008 and 2009 confirms a new introduction of the CA24 variant into the Americas from South-east Asia.

[Enteroviruses responsible for acute hemorrhagic conjunctivitis]. / Les Enterovirus responsables de conjonctivite aiguë hémorragique.

Acute hemorrhagic conjunctivitis (AHC) is an epidemic form of highly contagious conjunctivitis, characterized by conjunctival hemorrhages. The first AHC outbreak was described in 1969 in Ghana, West Africa, and was called Apollo disease, from the Apollo landing on the moon. This outbreak was caused by Enterovirus 70 (EV70) together with a Coxsackievirus A24 (CVA24v) variant, which are the major etiological agents involved in AHC outbreaks worldwide. AHC is known to be directly transmitted by close person-to-person contact or indirectly through soiled ophthalmological materials or unsafe recreational water. Recently, a possible airborne virus spread was suggested which could explain the high transmission rate of the disease. In the absence of a specific antiviral therapy, a rapid diagnosis of the causative agent is required to distinguish AHC due to enteroviruses from other ocular infectious diseases, for there are active drugs, or to quickly implement proper public health measures to limit the extension of the outbreak. However, virus identification remains difficult and time-consuming. Moreover, virological diagnosis is difficult to implement in developing countries where AHC has recently become a major problem for public health.

Modelling the transmission dynamics of acute haemorrhagic conjunctivitis: application to the 2003 outbreak in Mexico.

We model an outbreak of acute haemorrhagic conjunctivitis (AHC) using a simple epidemic model that includes susceptible, infectious, reported, and recovered classes. The model's framework considers the impact of underreporting and behaviour changes on the transmission rate and is applied to a recent epidemic of AHC in Mexico, using a fit to the cumulative number of cases to estimate model parameters, which agree with those derived from clinical studies. The model predicts a 'mean time from symptomatic onset to diagnosis' of 1.43 days (95 per cent CI: 1-2.5) and that the final size of the Mexican epidemic was underreported by 39 per cent. We estimate that a primary infectious case generates approximately 3 secondary cases (R0* = 2.64, SD 0.65). We explore the impact of interventions on the final epidemic size, and estimate a 36 per cent reduction in the transmission rate due to behaviour changes. The effectiveness of the behaviour changes in slowing the epidemic is evident at 21.90 (SD 0.19) days after the first reported case. Results therefore support current public health policy including expeditious announcement of the outbreak and public health information press releases that instruct individuals on avoiding contagion and encourage them to seek diagnosis in hospital clinics.

A recent epidemic of acute hemorrhagic conjunctivitis.

The coxsackievirus A24 variant was implicated in four of six major acute hemorrhagic conjunctivitis outbreaks in Singapore since its discovery in 1970. Hela cell suspension in 24-well flat-bottom tissue culture plates was a satisfactory alternative to monolayer cells grown in test tubes for virus isolation. Respiratory illness occurred in 20 of 98 patients with acute hemorrhagic conjunctivitis. Apart from conjunctival secretions, respiratory and oral transmission of coxsackievirus A24 variant would explain the rapid and extensive spread of acute hemorrhagic conjunctivitis during an outbreak.

Molecular epidemiology of a variant of coxsackievirus A24 in Taiwan: two epidemics caused by phylogenetically distinct viruses from 1985 to 1989.

In order to know the phylogenetic relationship and the route of transmission of a variant of coxsackievirus A24 (CA24v), an agent that caused four sequential outbreaks of acute hemorrhagic conjunctivitis from 1985 to 1989 in Taiwan, the nucleotide sequence variations in the virus-encoded proteinase 3C region (549 nucleotides) were studied with 19 isolates. The prototype strain (EH24/70), four isolates from Japan, and two isolates from Hong Kong were used for comparison. The nucleotide sequences of the Taiwan strains from the 1985-1986 and 1988-1989 epidemics were closely related within each epidemic, while they were more distantly related between strains from two epidemics. Phylogenetic analysis by the unweighted pairwise grouping method of the arithmetic average revealed that the 19 Taiwan isolates had diverged into two groups, 1985-1986 and 1988-1989 groups. The time at which these two groups diverged was estimated to be around May 1982, more than 3 years prior to the first appearance of the CA24v epidemic in Taiwan. On each occasion, the viruses caused a 2-year epidemic and then disappeared. The Taiwan isolates from 1985 to 1986 were closely related to the Japan isolates from 1985 to 1986 and the Taiwan isolates from 1988 to 1989 were phylogenetically close to the 1989 Japan isolates, indicating that Taiwan and Japan had two common-source outbreaks. However, none of the 1988 Taiwan isolates were phylogenetically close to the 1988 Japan or Hong Kong isolates. The evidence revealed that Taiwan has had two repeated but discontinuous introductions of CA24v since its first appearance in Taiwan in 1985. None of the other CA24v strains have been detected so far.

Spread of acute hemorrhagic conjunctivitis due to enterovirus-70: effect of air temperature and relative humidity on virus survival on fomites.

Enterovirus 70 (EV-70) has caused at least two pandemics and several major epidemics of acute hemorrhagic conjunctivitis (AHC) in the past 18 years, with the largest number of cases occurring in the coastal areas of the tropics. The exact means of its spread are not known, but fomites and hands contaminated by them are the most likely vehicles. We, therefore, tested EV-70 survival under different environmental conditions using stainless steel disks (1 cm diameter). Each disk received 10 microliter of the virus in phosphate-buffered saline (PBS). The disks were held at various temperatures with the relative humidity (RH) at the low (20 +/- 5%), medium (50 +/- 5%), high (80 +/- 5%), or ultrahigh (95 +/- 5%) level. The virus was eluted from the disks with tryptose phosphate broth and the eluate assayed in LLC-MK2cells. Inactivation rates (Ki), expressed as hourly loss of virus plaque forming units (PFU) in log10, were then calculated. At 20 degrees C, virus survival was proportional to the RH level, with the highest virus survival at the ultrahigh RH; at this level nearly 5% of the input virus was detectable even after 24 hr. Virus inactivation rates were only slightly higher at the ultrahigh RH when the temperature was raised to 33 degrees C or 35 degrees C. However, at 80% RH, increasing the temperature from 20 degrees C to 33 degrees C resulted in a dramatic rise in virus inactivation. The finding that high levels of RH prolong EV-70 survival on fomites may help explain the epidemiology of AHC resulting from EV-70.

Acute hemorrhagic conjunctivitis epidemic in Colon, Republic of Panama.

Acute hemorrhagic conjunctivitis has caused numerous outbreaks throughout Africa and Asia since it was first recognized in 1969 but did not involve the New World until 1981. This unprecedented outbreak reached Colon, Panama during August 1981 and by October 8,401 cases had been reported (14% of Colon's population). In October the Gorgas Memorial Laboratory and Ministry of Health conducted a survey in Colon to collect descriptive data on household living units, epidemiologic and clinical data from residents, and venous blood from all residents one year or older. The survey sampled 127 households and interviewed 608 people (1% of Colon's homes and 1% of the population). Overall 336 (55%) study subjects recalled having conjunctivitis. Disease rates differed according to residence; poor sectors of the city had 67% attack rates, lower class 52%, middle class 34%, and upper class 13%. Within each sector adults were more likely than children to be index cases, and communal bathrooms and household crowding were the most important risk factors for acute hemorrhagic conjunctivitis. Between 4-20% of people denying acute conjunctivitis had antibody to enterovirus 70 and may represent asymptomatic cases.