FIOCRUZ: Statement On Detection Of Zika Virus In Saliva and Urine

UPDATED (Feb 6th 0700hrs): Illustrating the risks of proclaiming `Eureka!',  Dr. Ian Mackay on his VDU blog has turned up at least one other instance of live Zika virus detected in urine in Urine found to contain infectious Zika virus...again...

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It isn't news that traces of the Zika virus can be found in urine or saliva, as viral RNA has been detected using RT-PCR testing since 2014 (see Detection of Zika virus in saliva), What hasn't been demonstrated - until now - is the presence of live Zika virus in these bodily fluids. 

The news hitting the AP and Reuters this morning has been the confirmation of live virus detection in saliva and urine by FIOCRUZ, Brazil's leading scientific research institute. 

While the implications of this finding are not yet clear, this will raise new questions about the routes of transmission of the virus from human-to-human.


Excerpts from the FIOCRUZ (Fundação Oswaldo Cruz) statement follows:

(translated)

Fiocruz detects the presence of zika virus with potential for infection in saliva and urine

Oswaldo Cruz Foundation (Fiocruz)

Foundation's pioneering study Oswaldo Cruz (Fiocruz), a body linked to the Ministry of Health, found the presence of active virus zika (with potential to cause infection) in saliva and urine samples. The unpublished evidence, which suggests the need to investigate the relevance of these alternative routes of virus transmission was observed by Flavivirus of Molecular Biology Laboratory of the Instituto Oswaldo Cruz (IOC / Fiocruz).

The studies were led by researcher Myrna Bonaldo, head of the Laboratory, in collaboration with infectious Patricia Brazil, the National Institute of Infectious Diseases Evandro Chagas (INI / Fiocruz). Samples were analyzed regarding two patients and samples were taken during the presentation of symptoms compatible with zika virus. Aliquots of the samples were placed in contact with Vero cells, which are widely used in studies of viral activity in the case of the flavivirus family, which belong to the Zika virus, dengue and yellow fever, among others.

Scientists have observed the CPE caused the cells: we observed the destruction or damage of cells, which proves the viral activity. The presence of the genetic material of zika virus was confirmed by RT-PCR in Real Time. It was also performed the partial sequencing of the virus genome. Laboratory diagnosis ruled out the presence of dengue virus and chikungunya - for these analyzes, we used the NAT Kit Discriminatory for Dengue, Chikungunya and Zika recently developed by Fiocruz. 

"It was known that the virus could be present in both urine and saliva. This is the first time we've demonstrated that the virus is active, ie, with the potential to cause infection, which opens new paradigms for understanding the transmission routes of the Zika virus. That answers an important question, however, the understanding of the epidemiological relevance of these potential routes of infection further studies "lies Myrna Bonaldo.

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The Lancet: Indentification Of Zika Virus Vectors

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The accepted wisdom - which is echoed in virtually every Zika story - is that the Aedes Aegypti mosquito is the primary vector for the virus. The Aedes Albopictus mosquito is also a suspected Zika vector, but its actual impact on the virus's spread is still unknown.

There are, however, more than 3,500 species of mosquito around the globe, and as least 175 can be found in the United States - including the two Aedes vectors (see map above) - and most have never been tested to see whether they can transmit Zika. 

A little over a week ago, in FIOCRUZ Researchers Investigate Other Possible Zika Mosquito Vectors, we looked at ongoing research looking to see if the Culex mosquito (common in the U.S. & in South America) might carry and transmit the Zika virus, but answers are still likely weeks away.


Last night The Lancet published a comment by Constância F J Ayres, an entomologist and research coordinator of the Culex/Zika research project at FIOCRUZ (Fundação Oswaldo Cruz), who outlines the lack of data on the Zika virus vectors and warns:

To assume that the main vector is A aegypti in areas in which other mosquito species coexist is naive, and could be catastrophic if other species are found to have important roles in Zika virus transmission.

The full comment (which is short) may be read at:

Identification of Zika virus vectors and implications for control

Constância F J Ayres

Published Online: 04 February 2016

DOI: http://dx.doi.org/10.1016/S1473-3099(16)00073-6

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UK: Aircraft Disinsection Ordered For Zika

Scheduled airline traffic around the world, circa June 2009 – Credit Wikipedia

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Although infected travelers are the most likely way for Zika (or Dengue, CHKV, or malaria) to make it from endemic regions to North America or Europe, aircraft and ships often carry unauthorized passengers; stowaway mosquitoes.


Up until now, `Airport Malaria'  has been the biggest concern, and the CDC even has a definition for it on their Malaria Transmission in the United States web page:

"Airport" Malaria

"Airport" malaria refers to malaria caused by infected mosquitoes that are transported rapidly by aircraft from a malaria-endemic country to a non-endemic country. If the local conditions allow their survival, they can bite local residents who can thus acquire malaria without having traveled abroad

In 2008, the American Society of Tropical Medicine and Hygiene warned:

Warmer climate changes in major U.S. cities with a large presence of international air traffic, such as New York and Los Angeles, seem to have created a more welcoming environment where these infected mosquitoes can survive. It begins with a mosquito that is transported during an international flight from a malaria-endemic region. Once the infected female mosquito leaves the aircraft, it can survive long enough to seek blood meals and transmit the disease to other humans within the airport. This type of international transmission creates an increased possibility for the reintroduction of not just malaria, but other detrimental diseases such as dengue and Chikungunya fever, into areas where they are not normally found.

Today the UK has announced that all aircraft inbound from Zika endemic regions will be required to undergo disinsection, a procedure which is currently done for flights from regions where malarial diseases are endemic.

 Planes returning from Zika areas to be sprayed with insecticide

From: Department of Health, Department for Transport and Public Health England

First published: 5 February 2016

All aircraft returning to the UK from countries currently affected by active Zika virus transmission will be sprayed with insecticide as part of a comprehensive government response to the disease.

On Monday, the World Health Organisation (WHO) declared the issue of microcephaly - which may be linked to Zika - a public health emergency of international concern.

As a precautionary measure, the government is asking airlines to ensure that disinsection (spraying with insecticide) takes place on all flights to the UK from countries with confirmed transmission of Zika.

Disinsection involves spraying a simple insecticide inside the aircraft to reduce the risk of passengers being bitten by any mosquitoes that could have entered the aircraft. It already occurs on the majority of flights from the region as a precaution against malaria.

The move is consistent with advice from WHO Europe. The type of mosquito that transmits the virus is extremely unlikely to survive and breed here given the lower temperatures in the UK.

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This practice, which is likely to become more common as Zika spreads, is not without its controversy. Although WHO weighed in on its safety 20 years ago (see below), some passengers have reported temporary ill effects or discomfort from the exposure. 

The choice, however, is to either to spray flights or allow potentially infected mosquitoes free international travel, with complementary in-flight blood meals thrown in as a bonus. 

The United States Transportation Department explains the procedures and the law:

Aircraft Disinsection Requirements
 
Overview

Disinsection is permitted under international law in order to protect public health, agriculture and the environment. The World Health Organization and the International Civil Aviation Organization stipulate two approaches for aircraft disinsection--either spray the aircraft cabin, with an aerosolized insecticide, while passengers are on board or treat the aircraft's interior surfaces with a residual insecticide (residual method) while passengers are not on board. Panama and American Samoa have adopted a third method, in which aircraft are sprayed with an aerosolized insecticide while passengers are not on board.

Although the Report of the Informal Consultation on Aircraft Disinsection sponsored by the World Health Organization (November 6-10, 1995) concluded that aircraft disinsection, if performed appropriately, would not present a risk to human health, the report also noted that some individuals may experience transient discomfort following aircraft disinsection by aerosol application.

Although few countries now require that aircraft be disinsected, most countries reserve the right to do so, and, as such, could impose a disinsection requirement should they perceive a threat to their public health, agriculture or environment. Accordingly, travelers are advised to check with their travel agent or airline reservations agent when booking flights. Listed below are representatives of airlines who are knowledgeable on disinsection requirements. 

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The UK appears to be the first major country to order the spraying of all flights from Zika affected regions, but other nations are no doubt considering similar steps.

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Hong Kong CHP Notified Of 19 Additional H7N9 Cases On Mainland

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As I mentioned yesterday (see H7N9 : Hunan Province Reports 2nd Case In Two Days), while official reporting of H7N9 cases from China has declined in reliability over the past year, recent media reports suggested there were roughly 20 unannounced cases on the Mainland, but where they occurred - and when - wasn't known.

Today, via a very brief (and data sparse) announcement from Hong Kong's CHP we are informed of 19 previously unannounced cases, with onsets going back as far as December 21st, hailing from three provinces (Zhejiang, Jiangsu & Fujian).

As always, surveillance only picks up those who are sick enough to be hospitalized, and lucky enough to then be tested for the virus. We know that mild and asymptomatic H7N9 infections do occur, but how often, is an open question.


This from Hong Kong's CHP.

CHP notified of additional human cases of avian influenza A(H7N9) in Mainland

The Centre for Health Protection (CHP) of the Department of Health (DH) today (February 5) received notification of 19 additional human cases of avian influenza A(H7N9) with onset of symptoms between December 21, 2015 and January 25 this year from the National Health and Family Planning Commission, and again urged the public to maintain strict personal, food and environmental hygiene both locally and during travel.

     The patients comprise 11 men and eight women aged from 42 to 91, four of whom (three men and one woman) have died. Eleven cases are from Zhejiang, five are from Jiangsu and three are from Fujian.

     "H5 and H7N9 avian influenza viruses can cause severe infections in humans. Due to the seasonal pattern, our risk assessment shows that the activity of avian influenza viruses is expected to remain at a high level in winter months. Human H5N6 and H7N9 cases have been recorded in the Mainland since this winter. Locally, birds positive for H5N6 were also detected this winter. The public, particularly poultry traders, travellers and those who may visit the Mainland in the coming Lunar New Year, should be highly alert. Do not visit poultry markets and farms. Avoid poultry contact. We will continue to monitor the regional and global disease situation," a spokesman for the CHP said.

     From 2013 to date, 702 human cases of avian influenza A(H7N9) have been reported by the Mainland health authorities.

     The DH's Port Health Office conducts health surveillance measures at all boundary control points. Thermal imaging systems are in place for body temperature checks on inbound travellers. Suspected cases will be immediately referred to public hospitals for follow-up.

     The display of posters and broadcast of health messages in departure and arrival halls as health education for travellers is under way. The travel industry and other stakeholders are regularly updated on the latest information.

     Travellers, especially those returning from avian influenza-affected areas with fever or respiratory symptoms, should immediately wear masks, seek medical attention and reveal their travel history to doctors. Health-care professionals should pay special attention to patients who might have had contact with poultry, birds or their droppings in affected areas.

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RESEP SETUP MAKARONI KEJU SUSU KHAS SOLO

RESEP SETUP MAKARONI

Olahan masakan dari pasta makaroni bisa mencoba resep setup makaroni khas Solo berikut ini. Olahan makaroni yang direbus empuk lalu dimasak berkuah susu bertabur keju ini konon digemari para bangsawan di masa kolonial Belanda. Menu kuliner enak asli Solo ini sering disajikan saat acara keraton surakarta maupun acara pesta pernikahan bahkan acara-acara hajatan lainnya.

Kuah susu membuat kuahnya menjadi lebih sedap dan creamy, makaroni yang empuk namun tidak lembek dengan taburan keju menjadi hidangan yang spesial beserta paduan daging ayam, kocokan telur, serta bumbu merica dan pala.

Perispan Bahan Bumbu Setup Makaroni

• 225 gram makaroni direbus cukup hingga empuk saja atau tidak sampai lembek
• 200 gram daging ayam dipotong dadu, lalu rebus hingga matang
• 3 buah sosis ayam dipotong korek
• 2 butir telur dikocok lepas
• 1 buah bawang bombay dipotong dadu
• 2 siung bawang putih
• 400 ml susu cair
• 1 liter air
• 1,5 sdt garam
• 1 sdt gula pasir
• 1 sdt merica
• 1 sdt pala bubuk
• 1 sdm margarin untuk menumis
• keju cheddar diparut seperlunya

Cara Membuat Setup Makaroni

1. Panaskan margarin, tumis bawang bombay dan bawang putih hingga harum. Masukkan daging ayam dan sosis, aduk rata kemudian tuang susu cair dan 1 liter air, beri garam, gula, merica, dan pala bubuk.
2. Aduk-aduk dan masak hingga mendidih, lalu masukkan makaroni yang sudah direbus dan kocokan telur, aduk-aduk dan masak sebentar hingga telur matang dan berbulir lalu matikan api. Angkat dan tuang setup makaroni dalam mangkuk saji serta sajikan hangat dengan taburan keju parut di atasnya.

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Eurosurveillance: Emergence of A(H1N1)pdm09 Genogroup 6B In India, 2015

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In February of 2015, in India’s H1N1 Outbreak, we looked at the persistent media reports of unusually severe H1N1 flu activity on the Indian subcontinent, and the insistence by India’s National Institute of Virology (NIV) and their National Centre for Disease Control (NCDC) that there were No mutations in H1N1.

A few weeks later, in MIT: Genetic Changes In A 2014 Indian H1N1pdm09 Virus,  we saw analysis of at least one flu isolate collected in 2014 that researchers said contained  `amino acid changes linked to enhanced virulence and are potentially antigenically distinct from the current vaccine containing 2009 (Cal0709) H1N1 viral hemagglutinin.

The entire report/commentary – which emphasizes the need for more robust and timely influenza surveillance and sequencing data -  can be accessed at:  Influenza Surveillance: 2014–2015 H1N1 “Swine”-Derived Influenza Viruses from India.

Fast forward to today, and the ECDC's Eurosurveillance has done just that.  

They've published an analysis of sequencing data from H1N1pdm09 isolates gathered in India during the first half of 2015. One that not only finds the rapidly rising Genotype 6B present but provides evidence of additional mutations as well.


This study also references the MIT study mentioned above. I've only excerpted a small piece, so follow the link to read it in its entirety.

Rapid communication

Emergence of influenza A(H1N1)pdm09 genogroup 6B and drug resistant virus, India, January to May 2015

M Parida ¹ , PK Dash ¹ , JS Kumar ¹ , G Joshi ¹ , K Tandel ¹ , S Sharma ¹ , A Srivastava ¹ , A Agarwal ¹ , A Saha ¹ , S Saraswat ¹ , D Karothia ¹ , V Malviya ¹

+ Author affiliations

Received:11 December 2015; Accepted:04 February 2016

---

To investigate the aetiology of the 2015 A(H1N1)pdm09 influenza outbreak in India, 1,083 nasopharyngeal swabs from suspect patients were screened for influenza A(H1N1)pdm09 in the state of Madhya Pradesh. Of 412 positive specimens, six were further characterised by phylogenetic analysis of haemagglutinin (HA) sequences revealing that they belonged to genogroup 6B. A new mutation (E164G) was observed in HA2 of two sequences. Neuraminidase genes in two of 12 isolates from fatal cases on prior oseltamivir treatment harboured the H275Y mutation.

An epidemic of influenza A(H1N1)pdm09, affecting over 39,000 persons and causing more than 2,500 deaths occurred in India in 2015 [1]. We show that genotype 6B strains forming two sub-lineages circulated during the outbreak.

Comparison of the sequences of six outbreak strains recovered in this work, to other published genotype 6B sequences, also reveals a unique combination of previously-reported mutations in the haemagglutinin (HA) gene. Two of the six sequences additionally display a E164G mutation in HA2, which has not been reported to date, moreover a N129D mutation in HA1 is observed for two sequences derived from patients with severe disease. Among strains analysed from 12 fatal cases on prior oseltamivir treatment, two harbour the H275Y mutation in the neuraminidase (NA) gene, which confers resistance to this antiviral.  

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The H275Y amino acid change, which confers resistance to Oseltamivir, has yet to take hold with the H1N1pdm09 virus, showing up in only 1%-2% of samples tested. That said, we have seen a few worrisome clusters of NAI resistant flu (see HERE & HERE), raising concerns that a resistant strain could evolve.


As mentioned previously, the A/H1N1pdm09 virus has remained remarkably stable since it first emerged as a pandemic strain in the spring of 2009.  But over time - and as all influenza A viruses do - the virus has picked up small genetic changes due to antigenic drift.

Most of these genetic changes go nowhere. They either offer no evolutionary advantage, or worse for the virus, they are detrimental to its survival.  

But occasionally - and through random chance -  a mutation (or set of mutations) provides the virus a biological advantage over its competitors; it infects its host more readily, transmits more easily, or replicates more efficiently.

The tendency is to become complacent with seasonal flu, and to worry only about the novel flu strains.  But the virulence of seasonal flu is not constant, and it is always possible for an old and familiar flu to learn new tricks. 

The changes observed in this study - and those recently reported by the Russians (see Week 5 Epi Report) - are intriguing and beg further investigation and study.  But it remains to be seen how much of an effect they will actually have on the virulence and future impact of the H1N1pdm09 virus.

 

Stay tuned. 

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H7N9 : Hunan Province Reports 2nd Case In Two Days

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Between Hong Kong CHP's tracking and the terrific work done by Sharon Sanders with FluTracker's H7N9 line listing, we've seen just over two dozen H7N9 cases announced by Chinese authorities since the fall flu season began.

While China earned praise during the first two H7N9 epidemic seasons for their daily updates on cases, for the past year we've seen the quality and quantity of reporting drop to abysmal levels. 

Earlier this week a Chinese media report alluded to there being nearly 20 unannounced cases (see China reports H7N9 since September last year 44 cases), but where they occurred - and when - isn't known.  


Over the past two days we've seen Xinhau reports announcing two recent H7N9 cases in Hunan Province (details below), and today's report indicates that Hunan has reported 5 cases since the 1st of the year.

A bit of a surprise because until yesterday, they'd only reported one.

 First an excerpt from yesterday's CHP announcement, followed by today's Xinhua report.

4 February 2016
CHP closely monitors additional human case of avian influenza A(H7N9) in Mainland 

     The Centre for Health Protection (CHP) of the Department of Health (DH) is today (February 4) closely monitoring an additional human case of avian influenza A(H7N9) in the Mainland, and again urged the public to maintain strict personal, food and environmental hygiene both locally and during travel.

     According to the Health and Family Planning Commission (HFPC) of Hunan Province, the 60-year-old male patient lived in Yongzhou, Hunan. 

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New human H7N9 case reported in central China

CHANGSHA, Feb. 4 (Xinhua) -- A new human H7N9 avian flu case was reported in central China's Hunan Province Thursday, bringing the total number of cases to five, local authorities said.

A 48-year-old man surnamed Xie died Thursday at a hospital in Yongzhou City, the provincial health and family planning commission said in a press release.

Xie, a native of Lingling District in Yongzhou, sought medication Wednesday and was diagnosed of human H7N9 avian flu. The immediate cause of his death was liver and kidney failures, the document said.

But it did not say how Xie contracted the illness.

Hunan Province has reported five cases of human H7N9 avian flu since the beginning of this year. Two of the patients have died and the other three are still receiving treatment, the document said.

Sporadic human H7N9 cases have been reported in Shanghai, Guangdong and Fujian. H7N9 is a bird flu strain first reported to have infected humans in March 2013 in China. It is most likely to strike in winter and spring.

While H7N9 activity appears lighter this winter in China than in previous years, the lack of dependable reporting out of China makes it very difficult to put a lot of faith in appearances.

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