Haemorrhagic Fever and Dengue Fever Notes

Haemorrhagic Fever

  • Definition:

    • Multi-system illness

    • Multiple organ dysfunction

    • Characterized by:

      • Fatigue

      • Generalized pain

      • Weakness and exhaustion

      • Fever (pyrexia)

      • Haemorrhage (bleeding)

Virus Classification

  • Baltimore Classification System:

    • Group I: dsDNA viruses

    • Group II: ssDNA viruses

    • Group III: dsRNA viruses

    • Group IV: Positive-sense, ssRNA viruses

    • Group V: Negative-sense, ssRNA viruses

    • Group VI: RNA reverse transcribing viruses

    • Group VII: DNA reverse transcribing viruses

Haemorrhagic Fever-Causing Viruses

  • Four main families:

    • Arenaviridae (Group V)

    • Filoviridae (Group V)

    • Flaviviridae (Group IV)

    • Bunyaviridae (Group V)

Arenaviridae

  • Characteristics:

    • Group V (- ssRNA)

    • Natural reservoir: rodents

    • Geographically restricted

    • Diagnosis: Clinical presentation, ELISA, RT-PCR, virus isolation

    • Prevention:

      • Isolation

      • Vaccine (Candid-1) for Argentina and Bolivian haemorrhagic fevers

    • Treatment:

      • Ribavirin

      • Supportive therapy

Specific Arenaviruses and Diseases

  • Lassa Fever

    • Causative agent: Lassa virus

    • Incubation period: 6-21 days

    • Symptoms: Cough, retrosternal pain, dyspnoea, pleuritis, unilateral or bilateral hearing deficit, seizures, tremors

    • Mortality rate: 1% overall, 15-20% in hospitalized patients, 50% during epidemics, 80% in 3rd trimester of pregnant women, 95% in foetus

    • Route of transmission: Inhalation of aerosols from excretions of rodents (Mastomys natalensis)

    • Other significant features: Endemic in West African countries, detection of rodents, complications (abortion, deafness, neurological disorders)

  • Argentine Haemorrhagic Fever

    • Causative agent: Junin virus

    • Incubation period: 7-14 days

    • Symptoms: Hypotension, infrequent urination, bradycardia, bleeding from all systems

    • Mortality rate: 3-30%

    • Transmission: Inhalation of aerosols from excretions of rodents (Calomys laucha and Calomys musculinus)

    • Restricted to the area of Junin, Argentina. Detection of rodents.

  • Bolivian Haemorrhagic Fever

    • Causative agent: Machupo virus

    • Incubation period: 7-16 days

    • Symptoms: Epistaxis, hematemesis (frequently present in the early stages), convulsions

    • Mortality rate: 3-30%

    • Transmission: Inhalation of aerosols, person-to-person, small abrasions and cuts in the skin.

    • Restricted to Bolivia. Detection of Calomys rodents aids diagnosis.

  • Venezuelan Haemorrhagic Fever

    • Causative agent: Guanarito virus

    • Incubation period: 7-16 days

    • Symptoms: Epistaxis, bleeding gums, hematemesis, melena, menorrhagia

    • Mortality rate: 30%

    • Transmission: Contact with excreta of two rodent species: Cane mouse (Zygodontomys brevicauda) and cotton rat (Sigmodon alstoni)

    • Confined area of central Venezuela. Detection of rodents.

  • Brazilian Haemorrhagic Fever

    • Causative agent: Sabia virus

    • Incubation period: 2-14 days

    • Symptoms: Multisystem bleeding, purpuric skin lesions

    • Mortality rate: 33%

    • Transmission: Inhaling infected aerosols

    • Confined to the area of Sao Paulo, Brazil

Filoviridae

  • Characteristics:

    • Group V (- ssRNA)

    • Natural reservoir: Unknown (possibly fruit bats or humans)

    • Diagnosis: Clinical presentation, ELISA, RT-PCR, virus isolation

    • Prevention:

      • Isolation

      • Minimizing contact

      • Barrier protection

    • Treatment: Supportive

      • Balancing electrolytes and lost body fluid

      • Maintaining patients’ oxygen and blood pressure in optimal level

    • Vaccine: Under development

Specific Filoviruses and Diseases

  • Ebola Haemorrhagic Fever

    • Causative agent: Ebola virus

    • Incubation period: 2-21 days

    • Symptoms: Abrupt onset of fever, blinding headache, joint and muscle pain, red eyes, skin rash, internal and external bleedings, bleeding from all systems, hiccup, diarrhoea, abdominal pain, multiple organ failure, lethargy, shock, death

    • Mortality rate: >90%

    • Transmission: Person-to-person, infected objects

    • Endemic in the tropics. Natural reservoir possibly human

  • Marburg Haemorrhagic Fever

    • Causative agent: Marburg virus

    • Incubation period: 5-10 days

    • Symptoms: Early stages - Abrupt fever, chills, general pain; Later - Maculopapular rash, liver failure, jaundice, shock, massive haemorrhage, multi-organ dysfunction

    • Mortality rate: Mild cases (23-25%), Severe cases (80-90%)

    • Transmission: Person-to-person, infected objects

    • Endemic in the tropics. Natural reservoir possibly bat

Flaviviridae

  • Characteristics:

    • Group IV (+ ssRNA)

    • Natural reservoir: Arthropods (mosquitoes)

    • Diagnosis: Clinical presentation, ELISA, RT-PCR, virus isolation

    • Prevention:

      • Vaccines (in some cases)

      • Isolation

      • Protective clothing

      • Insect repellent

      • Eradication of mosquitoes

    • Treatment: Supportive

      • Rebalance of lost fluid and electrolyte

      • Combating hypotension

      • Dialysis in acute renal failure

Specific Flaviviruses and Diseases

  • Yellow Fever

    • Causative agent: Yellow fever virus

    • Incubation period: 3-6 days

    • Symptoms: Early stage (3-4 days) - Pains, fever, jaundice; Period of remission - Fever and other symptoms alleviate, but 15% of infected individuals move to the third or the most dangerous stage within 24 hours; Period of intoxication - Seizures, multi-organ dysfunction, bleeding disorders, liver failure, kidney failure, coma, shock, death

    • Mortality Rate: 15-50%

    • Transmission: Insect bite

    • Disease is geographically distributed to 90% of Africa. The elderly are more prone to severe infection. Detection of mosquito Aedes aegypti for diagnosis.

  • Dengue Haemorrhagic Fever

    • Causative agent: Dengue viruses

    • Incubation period: 3-5 days

    • Symptoms: Headache, general body pain, localized retro orbital pain, increased vascular leakage, haemorrhagic phenomena, nausea and vomiting, leucopoenia

    • Mortality rate: Non-treated (20%), Treated (1%)

    • Transmission: Insect bite

    • World-wide distribution, but mostly in the tropics. Detection of vector Aedes aegyptiand Aedes albopictus. for diagnosis

Bunyaviridae

  • Characteristics:

    • Group V (- ssRNA)

    • Vectors: Ticks, mosquitoes, sand flies

    • Diagnosis: Clinical presentation, ELISA, RT-PCR, virus isolation

    • Prevention:

      • Isolation

      • Barrier protection

      • Reduced contact with infected animals

      • Self hygiene

      • Eradication of carrier

    • Treatment:

      • Ribavirin

      • Supportive therapy

Specific Bunyaviruses and Diseases

  • Rift Valley Fever

    • Causative agent: Rift Valley fever virus

    • Incubation period: 2-6 days

    • Symptoms: Mild - Asymptomatic, flue-like symptoms, meningitis-like symptoms; Severe - Ocular (0.5-2%) OR Meingoencephalitis (<1%) OR Severe haemorrhagic fever (>1%)

    • Mortality rate: Mild (1%), Severe (50%)

    • Transmission: Body fluid, tissue of infected animal, unpasteurized milk, mosquito bite, exposure to infected blood

    • Confined to the cattle & sheep raising regions e.g. Eastern and Southern Africa. No human-to-human transmission (except vertically in Sudan).

  • Haemorrhagic Fever with Renal Syndrome (HFRS)

    • Causative agent: Hantavirus

    • Incubation period: 7-14 days

    • Symptoms: Sudden onset of fever, blurred vision, acute shock, vascular haemorrhage, kidney failure

    • Mortality rate: Almost confined to Korean sub-continent, 5-15%

    • Transmission: Aerosols from rodents excreta, direct transmission of above with broken skin, mucous membrane of eyes, mouth & nose, rodent bites.

  • Crimean-Congo Haemorrhagic Fever (CCHF)

    • Causative agent: Crimean-Congo Haemorrhagic Fever Virus

    • Incubation period: 1-9 days

    • Symptoms: Early stages - Generalized pain, photophobia, non-bloody diarrhoea; Late stages - Confused, aggressive, bleeding from all systems/orifices, hepatomegaly, tachycardia, hepatorenal & pulmonary failure

    • Mortality rate: 30-50% (2nd week)

    • Transmission: Tick bite (ixodid tick), direct contact with infected tissues of livestock, direct contact with live livestock, human-to-human (nosocomial – hospital acquired)

    • Endemic regions in Asia, Africa, and Europe. Involvement with livestock, their fluid, tissue, and slaughter.

Dengue Fever

  • General Information:

    • Infects more than 50 million people annually, killing about 24,000 each year.

    • Acute, fever-causing disease, found in the tropics and Africa (geographically similar range to malaria).

    • Caused by one of four closely related virus serotypes of the genus Flavivirus, family Flaviviridae.

    • Transmitted to humans via the Aedes aegypti mosquito (Aedes albopictus).

  • Viral Genome:

    • Characterized by a small, enveloped virion.

    • Positive-sense, single-stranded RNA genome.

    • Causative agent of Dengue fever and Dengue haemorrhagic fever in humans.

  • Dengue Virus Types:

    • Difficult to examine because there are no laboratory or animal models of the disease.

    • Four distinct antigenic types (DEN-1, DEN-2, DEN-3 and DEN-4).

    • Indirect evidence suggests important biological differences among the viral genotypes.

  • Epidemiology:

    • Dengue is the most important arthropod-borne disease of humans.

    • DENV group consists of four antigenically closely related viruses: DENV1-4

    • DEN-1 and DEN-2 first identified during WWII in the Pacific; DEN-3 and DEN-4 isolated in Philippines in 1956

    • Over 2.5 billion people live in risk areas for dengue infection

    • 50-100 million cases annually, 20,000 deaths

    • Prevention and control: no vaccine yet available, vector control includes mosquito larvae source reduction and insecticide sprays

  • Clinical Features:

    • Most asymptomatic

  • Classical dengue fever: - Acute febrile illness, older children and adults, fever, rash, headache, muscle/joint pain, nausea, vomiting, rarely haemorrhage, convalescence of several weeks

  • Dengue haemorrhagic fever (DHF)/ Dengue shock syndrome (DSS): - severe form of the disease, fever, haemorrhage, vascular leakage, petechiae, hepatitis, risk factors: virus strain and previous infection with a different DEN serotype induced by Antibody Dependent Enhancement

    • No chemotherapy available, fluid replacement for DHF treatment

Antibody Dependent Enhancement

  • Mechanism:

    • First infection with a serotype of DEN

      • Mostly asymptomatic

      • May get DF, usually mild illness

      • Production of neutralizing antibodies to that serotype

    • Second infection with a different serotype

      • Antibodies to first serotype not neutralizing for second serotype

      • Antibodies attach with low affinity, enough to allow macrophages to take up virus (receptors for Fc)

      • Macrophages disseminate/spread virus though body

Transmission of Dengue

  • Cycles:

    • DENVs originated and are maintained in a forest (sylvan) transmission cycle involving canopy-dwelling Aedes spp. mosquitoes and primates

    • DENVs have fully adapted to humans and can also be maintained independently in an urban transmission cycle involving Ae. aegypti

    • Ae. aegypti is a highly domesticated (‘anthropophilic’) mosquito that has adapted to humans, preferring to feed on them and lay their eggs close to houses/ dwellings

Changing Epidemiology of Dengue

  • Global Emergence:

    • In the past 50 years, there has been a global emergence and re-emergence of epidemic dengue.

Reasons for Increased Dengue Activity

  • Factors:

    • Unplanned urbanization occurred in urban centres of SE Asia after WWII and in the Pacific and American tropics in the 1970s and 1980s

    • Lack of mosquito control in these newly populated centres

    • Advent of modern air travel increased movement of people, many of them infected and carrying DENV

    • Increased international commerce and trade has led to vector transport

    • Spread of vector due to changing environment (e.g., Europe)

Dengue in Australia

  • Historical Context:

    • Dengue occurred in Northern Territory, Queensland and New South Wales in the first part of the last century.

    • It disappeared following an epidemic in 1955 and reappeared in 1981 (DEN1)

    • Since then, regular epidemics in Northeast Qld

  • Current Status:

    • The distribution of the dengue vector Ae. aegypti is currently restricted to areas of Nthn Qld

    • Dengue is not considered endemic in Australia; epidemics arise from virus introduced by infected travellers.

Dengue Vaccine Development

  • Challenges:

    • DENV vaccine development has been underway for over 60 years.

    • The problems in development reflect the requirements of the vaccine to provide immunity to each the four serotypes (tetravalent).

  • Reasons for Tetravalent Requirement:

    • Infection with one serotype provides long-term protective immunity to this serotype, but not to others.

    • Severe DEN infection is associated with prior infection with a different serotype

      • Believed to be mediated by non-neutralising antibodies that remain from the first infection.

      • Mechanism of action is Antibody-Dependent-Enhancement (ADE), whereby pre-existing cross-reacting Abs facilitate entry of second infecting DENV into cells.

    • One or several serotypes can circulate simultaneously and serotypes can also change from one season to the next.

Vaccine Choices - Inactivated vs. Live-Attenuated

  • Inactivated Vaccine:

    • Require multiple inoculations for full protection

    • Elicit short-term immunity, requiring booster immunisation

    • Do not induce full range of immune responses observed during natural infection

    • Expensive to produce

  • Live-Attenuated (LAV) Vaccine:

    • Require 1-2 inoculations

    • Induce strong and long-lasting protective immunity

    • Immune response mimics natural infection

    • Relatively inexpensive to produce

Live-Attenuated Tetravalent Dengue Vaccine

  • Development:

    • The introduction of infectious cDNA technology has led to the development of LAVs for dengue and other flaviviruses.

    • For DENV, the most promising LAV is a chimeric virus which utilises an infectious clone of the Yellow Fever (17D) vaccine strain

      • YF17D is attenuated virus developed in the 1930s by serial infections of mouse embryos

      • Is one of the oldest vaccines and considered one of the safest and effective vaccines

      • ~400 million doses administered worldwide

    • In the chimeric vaccine (ChimeriVax-DEN), the genes encoding the YFV prM-Env have been replaced by those of DENV

      • Env protein is the dominant viral antigen, neutralizing antibodies

      • prM (pre-M protein, M is an envelope protein of unknown function) also induces neutralizing antibodies

Strategy for Constructing Chimeric Vaccine

  • Method

    • prM-Env genes of YF-17D removed by restriction digest

    • prM-Env genes of DENV amplified by RT-PCR

    • prM-Env genes of DENV ligated into YF-17D backbone

ChimeriVax-DEN Performance Evaluation/Trial Results

  • Preclinical Results

    • Cell culture: Replicated to high titres

    • Neurovirulence in mice: Non-neurovirulent

    • Rhesus monkeys: Immunogenic (full seroconversion), Low level viraemia

    • Replication in Ae. Aegypti: Reduced growth, loss of virus dissemination

  • Clinical Phase I Results

    • Safety and immunogenicity: Consistent with YF17D vaccine, pre-immunity to DENV does not affect response, pre-immunity to YFV enhanced DENV antibody response, 100% seroconversion

Haemorrhagic Fever Outbreak in Herat, Afghanistan - Case Study

  • Location:

    • Gulran, Herat, Afghanistan

    • Shoor Ab Village

    • Population: 500 people

    • Occupation: animal husbandry, farming

  • Epidemiology:

    • Number of infected individuals: 26

    • Symptoms: Fever, haemorrhagic manifestations from all systems/orifices, dizziness, lethargy, headache, petechial rash, hepatomegaly and liver failure, renal failure, pulmonary failure

    • Age: 22-70, mainly in 40s

    • Index cases:

      • Two shepherds

      • One butcher

      • One farmer

    • Other cases:

      • 13 - family members of the index cases

      • 1 - Mullah of the community

      • 8 - shepherds, farmers, teachers in the village

  • Timeline:

    • May 2000: initiation of outbreak

    • DoH initiated investigation 2 weeks after 1st cases

    • June 2000: WHO was called for their assistance

      • A group of experts from WHO arrived in the region <10 days after DoH commenced investigation

      • An isolation room setup within Herat Hospital, equipped

      • Samples from all patients were sent to NIV

      • Treatment and control measures were established 10 days post DoH investigation

      • Training seminars for health care staff

    • July 2000: NIV reported causative agent not identified

    • July 2000: Another group of experts arrived in Herat

  • Post Outbreak Summary:

    • The causative agent: not identified

    • Control measures: appropriate BUT delayed commencement

    • Treatment: Ribavirin, supportive BUT delayed commencement

    • The year 2000 drought in the country killed all livestock and this contributed to eradication

    • Mortality: n=15/26 (58%)

Factors Contributing to High Mortality Rate

  • Reasons:

    • Lack of awareness of healthcare workers of haemorrhagic fever

    • Taliban Government: underdeveloped health system

    • No reporting of suspected cases following the first cases

    • DoH only initiated investigation two weeks late

    • Control measures and treatment: 25 days late

    • Public lack of awareness

Why Causative Agent was Not Identified?

  • Possible Errors:

    • Error in sample collection?

    • Error in transporting?

    • Inappropriate transport media?

    • Time elapsed between sample collection and laboratory examination

Virus Family, Viruses, Epidemiology, Human-to-Human Transmission, Clinical Features

  • Arenaviridae

    • Lassa virus: Endemic in West African Countries, NO Human Transmission Rodents

    • Junnin virus: Argentina, NO Human Transmission

    • Machupo virus: Bolivia, YES Human Transmission

    • Guanarito virus: Venezuela, NO Human Transmission

    • Sabia virus: Brazil, NO Human Transmission

  • Filoviridae

    • Ebola virus: Tropics, YES Human Transmission Fruit bat?, Human?

    • Marburg virus: Tropics, YES Human Transmission

  • Flaviviridae

    • Yellow fever virus: 90% of Africa, NO Human Transmission Mosquito

    • Dengue virus: Worldwide, but mostly tropics, NO Human Transmission

  • Bunyaviridae

    • RVF virus: South African countries, Saudi Arabia, NO Human Transmission Mosquito

    • CCHF virus: Asia, Africa, Europe, YES Human Transmission Tick, Livestock

    • Hantavirus: Korea, South America, RARELY Human Transmission Rodents

Causative Agent of Herat Outbreak

  • Epidemiological Studies:

    • Livestock involvement

    • Crimean-Congo Haemorrhagic Fever (CCHF) is endemic in Iran and Pakistan

    • Refugees coming from endemic area

    • Livestock imported from endemic area

  • Clinical Features:

    • Dizziness

    • Lethargy

    • Hepatomegaly

    • Pulmonary failure

    • Haemorrhagic manifestations from all systems/orifices

CCHF Identified in Herat 2008

  • Second Outbreak:

    • August 2008

    • Same area, same clinical features

    • CCHFV isolated from patients’ samples

    • 10 cases leading to 3 deaths

    • Treatment, control measures established within two days

    • No new cases and no further deaths

  • Eradication:

    • The disease eradicated in less than two weeks

Summary of Haemorrhagic Fevers

  • HF is a multi-system illness with bleeding and fever

  • Four families of viruses can cause HF

  • Severity, mortality rate and intervention differ according to the causative agent

  • Likely that the original outbreak in 2000 was caused by CCHF

  • Early diagnosis, treatment and prevention are essential in controlling the outbreak and preventing the spread of the disease

Here are the answers to your questions based on the provided notes:

  1. Which 4 virus families are involved with causing haemorrhagic fever?

    • Arenaviridae

    • Filoviridae

    • Flaviviridae

    • Bunyaviridae

  2. List the viruses in each family causing haemorrhagic fever

    • Arenaviridae: Lassa virus, Junin virus, Machupo virus, Guanarito virus, Sabia virus

    • Filoviridae: Ebola virus, Marburg virus

    • Flaviviridae: Yellow fever virus, Dengue viruses

    • Bunyaviridae: Rift Valley fever virus, Hantavirus, Crimean-Congo Hemorrhagic Fever Virus

  3. List the types of clinical features caused by dengue.

    • Headache

    • General body pain

    • Localized retro orbital pain

    • Increased vascular leakage

    • Hemorrhagic phenomena

    • Nausea and vomiting

    • Leucopoenia

  4. What is “antibody dependant enhancement”?

    • ADE occurs when a second infection with a different serotype of dengue virus happens.

    • Antibodies from the first infection are not neutralizing for the second serotype.

    • These antibodies attach with low affinity, allowing macrophages to take up the virus.

    • Macrophages then disseminate the virus throughout the body.

  5. Give reasons for the increase in spread of dengue.

    • Unplanned urbanization in urban centers of Southeast Asia after WWII and in the Pacific and American tropics in the 1970s and 1980s.

    • Lack of mosquito control in these newly populated centers.

    • Increased modern air travel, increasing movement of infected people.

    • Increased international commerce and trade, leading to vector transport.

    • Spread of the vector due to changing environment.

  6. What is the major issue with dengue vaccines?

    • The dengue vaccine must provide immunity to each of the four serotypes (tetravalent), as severe dengue infection is associated with prior infection with a different serotype.

  7. How is the ChimeriVax-DEN vaccine made?

    • The ChimeriVax-DEN vaccine is a chimeric virus that utilizes an infectious clone of the Yellow Fever (17D) vaccine strain.

    • The genes encoding the Yellow Fever virus prM-Env proteins are replaced by those of the Dengue virus.

  8. What were the contributing factors causing the mortality rate in the 2000 Gulran outbreak

    • Lack of awareness of healthcare workers of hemorrhagic fever.

    • Underdeveloped health system due to the Taliban government.

    • No reporting of suspected cases following the first cases.

    • Delayed investigation by the Department of Health.

    • Delayed control measures and treatment.

  9. How was dengue and yellow fever excluded as possible causes of the 2000 Gulran outbreak?

The causative agent was not identified in the initial investigation by NIV (National Institute of Virology). Samples were sent from patients in the outbreak but came back without any determination of the cause. This would also include testing to see if it was Dengue or Yellow Fever.