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Indonesian H5N1 Bird and Human Sequences Do Not Ma

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Jhetta View Drop Down
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    Posted: July 03 2006 at 4:55pm
Commentary
 

Indonesian H5N1 Bird and Human Sequences Do Not Match

Recombinomics Commentary

July 3, 2006

The recent meeting in Jakarta on human H5N1 in Indonesia raised additional question on the origin of the human H5N1.  The meeting included a phylogenetic tree of HA sequences from human, cat, and bird isolates.  The human and cat sequences formed a separate branch which did not include avian sequences.  The names of those isolates are listed below and are on lower branch of the phylogenetic tree.

The report in Jakarta also listed those bird flu isolates with their novel cleavage site RESRRRKKA.  All such isolates are on the lower branch of the tree which has all human isolates in Indonesia other than the Kao cluster and the second reported case.  Included in the lower branch is a cat sequence, feline/IDN/CDC1/06.

The slide that listed the amantadine resistant isolates included the age, gender, date, and location of the human case.  These cases extend from the first reported case in July of 2005, IND/05/05, through cases in May 2006, IDN/554H/06 and IDN/557H/06.  The cases include isolates in and around Jakarta as well as a case in East Java.  Thus; although the human cases have been isolated for almost a year and throughout the island of Java, none of the human isolates match a bird isolate.

A recent phylogenetic tree has
19 bird and two swine H5N1 isolates from isolates across Indonesia.  Thus, the number of non-human isolates in Indonesia now exceeds fifty and only the cat isolate falls on the human branch which has 20 H5N1's isolated in Hong Kong (in addition to corresponding isolates by the CDC).

The failure of the twenty human isolates to match the 50 avian isolates suggests birds are not the source of H5N1 in most of the human cases in Indonesia.  In spite of this failure to match,  WHO updates continue to cite bird deaths in some sort of association with the human cases. 
 
However, H5N1 is widespread in birds in Indonesia, and the sequences indicate that the correlation between human and avian H5N1 does not exist, based on the sequences described in the Jakarta meeting and released in the form of phylogenetic trees.

The failure to match human and avian sequences raise serious credibility issues regarding WHO updates.  The two cased listed above were described in the Who
update of May 29

One newly confirmed case is an 18-year-old man from East Java Province. He developed symptoms on 6 May and was hospitalized on 17 May. He is now recovering. The investigation found a history of exposure to dead chickens in his home within the week prior to symptom onset. No further cases of influenza-like illness have been identified during the investigation and monitoring of his close contacts.

An additional case occurred in a 39-year-old man from West Jakarta. He developed symptoms on 9 May, was hospitalized on 16 May, and died on 19 May. The investigation determined that the man cleaned pigeon faeces from blocked roof gutters at his home shortly before symptom onset. No further potential source of exposure was identified.

The isolate from the 18M, IDN/554H/06, and 39M, IDN/557H/06.were both similar to the first human Indonesian isolate IDN/05/05 and have the same RESRRKKR cleavage site.  No reported avian isolates from 2003 to 2005 have this cleavage site or sufficient similarity to be placed on the same branch of the phylogenetic tree, yet the updates continue to use dead or wild birds as a likely cause of the human infections.  WHO is well aware of the failure to find match bird isolates, yet bird contacts are used in updates and Indonesia uses history of contact with dead or dying birds as a criteria for H5N1 testing.

These approaches fail to address the true origins of H5N1 infections in Indonesia and create a climate of deception.  The WHO and Indonesian approaches for surveillance and containment of H5N1 in humans in Indonesia are increasing causes for concern.

Indonesian human isolates on lower branch of HA phylogenetic tree

IDN/175H/05
IDN/07/05
IDN/160H/05
IDN/557H/06
IDN/177H/06
IDN/554H/06
IDN/05/05
IDN/195H/05
IDN/298H/06
IDN/542H/06
IDN/567H/06
IDN/321H/06
IDN/341H/06
IDN/245H/06
IDN/304H/06
IDN/239H/05
IDN/292H/06
IDN/286H/06
IDN/283H/06
IDN/282H/06

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Post Options Post Options   Thanks (0) Thanks(0)   Quote Guests Quote  Post ReplyReply Direct Link To This Post Posted: July 03 2006 at 5:02pm

  Jhetta,

Would you please "translate" this for us?  I don't understand.  Blushy 2





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He is basically saying based on the H5N1 sequences that he has been able to examine... it looks like the humans who contracted H5N1 did not get it from birds!
 
My bet would be pigs!
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Have there been reports of pigs or other animals in Indonesia dying? If not then asymptomatic animals passing deadly H5N1 to humans?  So, there is some unidentified source of the virus that killed all those people? Oh boy!

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hmm........................

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Originally posted by Jhetta Jhetta wrote:

He is basically saying based on the H5N1 sequences that he has been able to examine... it looks like the humans who contracted H5N1 did not get it from birds!
 
My bet would be pigs!
 
If it is in fact pigs that have been the transfer point to humans....it might be short term "good news" --  but in the medium / long term it is very bad news.
 
The "good" news is that pigs don't fly.  H5N1 cannot migrate via pigs.  Thus--short term, this limitation would logically limit H5N1's spread in a form that infects humans.
 
The bad news is...swine happen to be (apparently) the only mammalian "mixing pot" for H5N1 -- they are susceptiable to both human flu AND H5N1.  When  both viruses have infected a swine host, the virii can reassort--recombine DNA material  between them at a very rapid pace....making for very rapid mutations in H5N1. 
 
One of the logically possible mutations, given this scenario, is H5N1 could easily gain the easy transmissibility of the common flu into humans, while retaining its lethality. 
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Originally posted by 4thegirls 4thegirls wrote:

Have there been reports of pigs or other animals in Indonesia dying? If not then asymptomatic animals passing deadly H5N1 to humans?  So, there is some unidentified source of the virus that killed all those people? Oh boy!

 
Here is some background info... H5N1 was detected long ago in indonesia's pigs; they can be asymptomatic and they have repeately failed to cull pigs or poultry!
 
An internal audit at Indonesia's agriculture ministry has found suspected corruption in the provision of vaccines to fight a bird flu outbreak.
http://www.abc.net.au/ra/news/stories/s1477919.htm
 
THE WASHINGTON POST

Failed Indonesian bird flu response concerns experts
http://www.abc.net.au/am/content/2006/s1578301.htm
 
 
Nature report on H5N1 virus in pigs in Indonesia
 
HIGHLY PATHOGENIC AVIAN INFLUENZA IN INDONESIA
Follow-up report No. 8 (H5N1 infection detected in pigs)
 
HIGHLY PATHOGENIC AVIAN INFLUENZA IN INDONESIA (2006 Avian) Follow-up report No. 12
http://www.oie.int/eng/info/hebdo/AIS_21.HTM#Sec2
 

 
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Post Options Post Options   Thanks (0) Thanks(0)   Quote Guests Quote  Post ReplyReply Direct Link To This Post Posted: July 03 2006 at 6:28pm
jhetta, have you seen the report from channel 4 news? its on www.microbes
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Originally posted by SHERRIE SHERRIE wrote:

jhetta, have you seen the report from channel 4 news? its on www.microbes
No... not yet.. I do not watch much TV... is it good?
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  Jhetta,  I appreciate the help!   You’re The Best





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Originally posted by 4thegirls 4thegirls wrote:

  Jhetta,  I appreciate the help!   You’re The Best
 
Thanks 4thegirls... it is nice to be appreciated!  Have a great 4th!
 
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jhetta go to microbes.info and click on flu then on channel 4 news it was from today i thought it was a little alarming. i still dont know how to post stuff so thats why i asked you cause your smart. thanks sherrie
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A recent phylogenetic tree has 19 bird and two swine H5N1 isolates from isolates across Indonesia.  Thus, the number of non-human isolates in Indonesia now exceeds fifty and only the cat isolate falls on the human branch which has 20 H5N1's isolated in Hong Kong (in addition to corresponding isolates by the CDC).
 
I think the cats are the spreaders, I read a study not to long ago (sorry I can't remember where) about the possibility of cats being a 'mixing pot' for avian influenza. Conventional thought is that the pig is the mixer and they are, but new thinking is that the cat is too. Since the human sequences more closely match the cat than the pig or the bird sequences, I hope someone is rounding up some cats.
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Originally posted by SHERRIE SHERRIE wrote:

jhetta go to microbes.info and click on flu then on channel 4 news it was from today i thought it was a little alarming. i still dont know how to post stuff so thats why i asked you cause your smart. thanks sherrie
 
Is this it?  Sounds accurate to me... still watching the video... thanks... best news feed I have seen so far!  Great find Sherrie... I suspect they are trying to keep the news under raps as much as they can.  This is probably even a little conservative...
 
 
 
Special Reports 
 
Indonesian crisis?

Published: 3 Jul 2006
By: Ian Williams


Dead chickens litter the streets, but has Bird Flu begun to spread between humans too?

>>Watch the report

It's registered more deaths from bird flu than any other country this year - just today tests confirmed a five year old boy became Indonesia's fortieth victim of the disease.

Scientists already believe there could have been limited human to human transmission of the virus when six members of the same family died in North Sumatra.

However they still don't know much about how the disease spreads among chickens.

Our Asia correspondent Ian Williams has travelled to West Java - where a team of veterinary experts from around the world are racing to set up a surveillance system to understand how bird flu has spread.

Their early findings aren't encouraging.
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yes thats it, see i knew you were smart. i find things but thats all im good for. i thought it was under control in indo. since no recent reports until today.
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Originally posted by rodin33 rodin33 wrote:

A recent phylogenetic tree has 19 bird and two swine H5N1 isolates from isolates across Indonesia.  Thus, the number of non-human isolates in Indonesia now exceeds fifty and only the cat isolate falls on the human branch which has 20 H5N1's isolated in Hong Kong (in addition to corresponding isolates by the CDC).
 
I think the cats are the spreaders, I read a study not to long ago (sorry I can't remember where) about the possibility of cats being a 'mixing pot' for avian influenza. Conventional thought is that the pig is the mixer and they are, but new thinking is that the cat is too. Since the human sequences more closely match the cat than the pig or the bird sequences, I hope someone is rounding up some cats.
 
Would not be surprised by that either.... however pigs are suseptible to both avian and human virus... and they have both 2.3 and 2.6 receptors.
 
Will have to look into cat receptors... there are documented cases of wide spread infections of tigers at a zoo and cats infected in germany etc... will post that info later... I have to get ready for a dinner date!
 
 
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It would be nice if Niman could look at current data from cats, pigs, humans in all areas where people have died in recent weeks/months.
 
Leaning towards cats... at this point
 
Commentary
 

Indonesian H5N1 Swine and Bird Sequences Are Similar

Recombinomics Commentary

July 4, 2006

The recent meeting on human H5N1 bird flu in Indonesia included phylogenetic trees of HA and M genes.  The HA tree identified two separate groupings of the human isolated, color coded in green.  The Karo cluster was related to avian isolates.  This group is shaded in pink.  Another human isolate, IDN/06/05 is the second confirmed Indonesian human case and it is located on a nearby branch.  However, the remaining human cases formed a separate branch, located at the bottom of the tree.  The only non-human isolate on this branch is an H5N1 isolate from a cat, feline/IDN.CDC1/06.  Like the other isolates on the branch, it had a novel cleavage site, RESRRKKR.

The presence of the novel cleavage site in human and cat isolates raised the possibility that the source of the human infections was mammalian rater than human.  This first human isolate in Indonesia was in Banten in July 2005.  At the time H5N1 was found in swine in Banten and a recent presentation included two H5N1 swine isolates from Banten, as well as 19 chicken isolates from locations throughout Indonesia.  These are listed in a
phologenetic tree and show that the swine sequences are similar to bird sequences in Indonesia.  Moreover, the swine sequences have the common HA cleavage site RERRRKKR, which is in all but two 2003 isolates on the phylogenetic tree.  Similarly, most of the Indonesian bird isolates in Indonesia are RERRKKR.

These data indicate the H5N1 in the swine in Banten are not the source of the H5N1 in humans in and around the Jakarta area.  In the past year, the human isolates continue to point away from an avian source.  There are now over 50 avian isolates collected between 2003 and 2005 in Indonesia.  There are a few isolates with novel cleavage sites of RERRRIKR, RERRRIKK, RERRRK_R, and
GERRRKKR.  However, the vast majority of the bird sequences are RERRRKKR.  In contrast, other than the Karo cluster and the second human isolate in Indonesia, all other human isolates form a separate branch on the tree and virtually all have an RESRRKKR cleavage site.

The failure to find a matching source for these sequences, other than one cat isolate, is cause for concern. This failure raises serious surveillance issues in Indonesia and  raises doubts concerning transparency in WHO updates, which continue to point toward bird interactions, but fail to identify H5N1 in contact poultry and fail to find matching sequences in a growing number of bird isolates.

The human sequences continue to evolve, yet the sequences of the isolates remain sequestered in a WHO private database.  The only HA sequence made public is the sequence from the first human case confirmed in Indonesia.  That sequence was placed in the WHO database on
August 1, 2005.  The phylogenetic tree lists human sequences from over 25 individuals. 

The time for release of these data has
past.

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Post Options Post Options   Thanks (0) Thanks(0)   Quote Jhetta Quote  Post ReplyReply Direct Link To This Post Posted: July 04 2006 at 2:28am
Here is the info I promised regarding expanding host range... i.e. cats
 
I emailed key parts of this info to the labs that will be screening both human and animals for H5N1 infections in San Diego so they will be aware that it does not just infect birds.
 
 
Bird flu: Don’t put the cat among the chickens, warn Dutch team

PARIS - A leading team of European virologists has appealed for health authorities to step up vigilance about household pets, saying cats and possibly dogs too are at risk from bird flu.

Albert Osterhaus and colleagues at the Erasmus maskmanal Centre in Rotterdam, the Netherlands, say that the World Health Organisation (WHO) and other agencies have under-estimated the risk from cats in their campaign against avian influenza.

They point to several documented cases in Thailand, Indonesia and Germany in which domestic cats, farmyard cats and zoo felines have fallen sick or died after eating H5N1-infected chickens or wild birds, including the death of 147 tigers at a Thai zoo in 2004.

And they say that lab tests they have conducted prove that cats can catch the virus in several ways -- either from eating infected chicks, through contact with infected birds or through virus administered directly into the respiratory tract.

“The available evidence, albeit incomplete, suggests that cats are more than collateral damage in avian flu’s deadly global spread and may play a greater role in the epidemiology of the virus than previously thought,” the Dutch experts say in a commentary published on Thursday in Nature.

The Osterhaus team acknowledge that no-one knows if an infected cat can pass on H5N1 to humans.

Just as unknown is whether the animal, by harbouring the virus, can help it to mutate into a pandemic form -- a pathogen that is not only lethal for humans but contagious, too.

But, they say, this risk cannot be ruled out, and precautions should thus be incorporated into the guidelines of the WHO, the Food and Agricultural Organisation (FAO) and the World Organisation for Animal Health (OIE).

They recommend that steps be taken to prevent contact between cats and infected birds or their droppings; cats suspected of such contacts should be quarantined; and in temperate climates where there has been an outbreak of bird flu, cats should be kept indoors.

Surveillance should also be boosted for any sign of the bird flu virus among dogs, foxes, weasels, stoats and seals, as “we now know that H5N1 virus has the ability to infect an unprecedented range of hosts, including carnivores,” the commentary adds

 
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H5N1 In Cats and Dogs

Introduction


At the end of February 2006 highly pathogenic avian influenza (HPAI), caused by the H5N1 virus was detected in a domestic cat found dead on the northern island of Ruegen, Germany. Since mid-February, over 100 birds have died on this island and tests confirmed H5N1 infection. Also in Asia, cats and other felidae are occasionally found to be infected with H5N1 since the start of the poultry epidemic end 2003. Experimental studies have shown that the domestic cat can become infected with the virus and that cat to cat transmission is possible in principle. Serological studies in several Asian countries suggest that dogs may also contract the H5N1 infection. Countries in Europe have advised owners of pets living near H5N1 wild bird foci to keep cats indoors and dogs on a leash when taken for a walk.

These recent events lead to many questions by the public and pet owners to which the veterinary profession has to respond. In addition, there may be exposure of pet owners and veterinarians. For example, when animals infected with H5N1 (eg birds, dogs and cats) are brought to the veterinary clinic. Important are also the contribution veterinary practitioners can make in the surveillance of the disease for the presence of the H5N1 infection.

This section provides information for the general public and professionals about the risk of cats contracting H5N1 virus and the role of cats in the spread of avian influenza H5N1.


Background

http://www.fao.org/ag/AGAinfo/subjects/en/health/diseases-cards/avian_cats.html#back back


During a H5N1 outbreak in poultry in 1997 in Hong Kong, the first clinical human cases of this sub-type were reported with several fatalities. From the end of 2003 to date (March 2006) 173 people have been confirmed infected with the H5N1 virus of which 93 have died. Except for 1 case, human-to-human transmission has probably not occurred. Although H5N1 is relatively common to wild birds and poultry, humans and other mammals are also at risk of HPAI infection. Highly pathogenic avian influenza in poultry is of growing concern due to the current geographic extent comprising Asia, Africa and Europe showing potential for pandemic spread. The virus is highly contagious and already over 200 million domestic birds have either been culled or died of the disease. Table 1 shows the timeline for avian influenza in cats and other felidae.


Timeline of (H5N1) avian influenza in cats and other felidae (and civets)

1970s & 1980s

Research revealed that infection of domestic cats with influenza A subtypes H3N2 from humans, H7N3 from a turkey, and H7N7 from a harbor seal (Phoc vitulina) produces transient virus excretion and a temporary increase in body temperature but did not induce any other clinical signs of disease.

December 2003

Two leopards and two tigers died at a zoo in Thailand after feeding on chicken carcasses. Investigation confirmed H5N1 in tissue samples from all 4 animals. This was the first report of influenza causing disease and death in big cats.

September 2004

Research shows that domestic cats experimentally infected with H5N1 develop severe disease and can spread infection to other cats.

October 2004

A H5N1 outbreak in zoo tigers in Thailand reportedly fed on chicken carcasses. Eventually, 147 out of the population of 441 tigers died or had to euthanized for animal welfare reasons.

June 2005

Tests on three civets that died late June 2005 in Viet Nam revealed H5N1, marking the first infection of this species with the virus. These endangered Owston’s palm civets were raised in captivity; source of infection is still unknown.

October 05 February 06

FAO field veterinarians report unusual high cat mortality in Iraq and Indonesia in the vicinity of H5N1 outbreaks in poultry.

28 February 2006

H5N1 confirmed in a cat on the Baltic Sea island of Ruegen (Germany). Over 100 wild birds had been found dead on the island during previous weeks.



General Information

http://www.fao.org/ag/AGAinfo/subjects/en/health/diseases-cards/avian_cats.html#back back


Role of cats in virus transmission
Research has shown that domestic cats may die from H5N1 virus. Also horizontal transmission has been proven. However, it is unlikely that cats play a role in the natural transmission cycle of H5N1 viruses. Cat infections occasionally occur in association with H5N1 outbreaks in domestic or wild birds, e.g. when cats feed on infected birds. Experimental/infected cats shed the virus via the respiratory and intestinal tract, and may therefore transmit the virus to other cats. Naturally infected cats are thus in theory, able to spread the virus

In areas where H5N1 Infected wild birds are reported it can not be excluded that cats become infected. Although most wild birds infected are waterfowl, not normally the species cats interact with, H5N1 is potentially infectious to numerous other bird species and it can not be ruled out that passerines or pigeons which do interact with cats get infected

In areas where poultry is infected with H5N1 there is a risk that cats become infected with H5N1 through contact with infected poultry or their faeces. Anecdotal reports support the notion that contact with infected poultry (faeces and eating infected carcasses) forms a source of infection for cats. Cats probably have little or no contribution to the spread of the disease because the number of infected poultry is much higher than the number of infected cats; poultry shed much more virus than cats. Nevertheless, cats may play a role in the spread of the virus to other animals. Report to the local veterinary authority any evidence of significant animal mortality both wild and domestic.

Theoretically there is a possibility that cats transmit infection to humans. However, given the risk that cats become infected with HPAI is low, the risk to human infection is therefore limited.

The role of stray cats
Due to their greater mobility, stray cats could spread the disease into new areas. If infected, stray cats may become a source of contamination to poultry and mammals, including humans.

The role of other mammals
The ability of catching the H5N1 virus is not restricted to cats. Reports show infection in tigers, leopards and civets. Also dogs and pigs may become infected with the virus. Given the broad host spectrum of the H5N1virus, the possibility that also other wild or domesticated mammals including seals, mustelidae or furbearing animals, become infected by contacting infected animals is present. All carnivores could become infected through eating infected poultry or infected wild birds.

Recommendations
Areas where H5N1 HPAI has been diagnosed or is suspected in poultry or wild birds:

Report to the local veterinary authority any evidence of significant bird mortality both wild and domestic
Be especially vigilant for any dead or sick cats and report such findings to the local vet
Make sure contact between cats and wild birds or poultry (or their faeces) is avoided and/or keep cats inside
If cats bring a sick or dead bird inside the house, put on plastic gloves and dispense of the bird in plastic bags for collection by local veterinary animal handlers
Keep stray cats outside the house and avoid contact wit them
If cats show breathing problems or nasal discharge, a veterinarian should be consulted
Do not touch or handle any sick-looking or dead cat (or other animal) and report to the authorities
Wash hands with water and soap regularly and especially after handling animals and cleaning their litter boxes or coming in contact with faeces or saliva
Dogs can only be taken outside the premises if kept restraint
Do not feed any water birds
Disinfect (e.g. with bleach 2-3 %) cages or other hardware with which sick animals have been transported or been in contact with.
Wash animal blankets with soap or any other commercial detergent


Information for veterinarians

http://www.fao.org/ag/AGAinfo/subjects/en/health/diseases-cards/avian_cats.html#back back


Avian influenza in other animal species
Hosts: Wildbird hosts for H5N1 in order of importance are probably Anatidae (ducks, geese, swans), Charadriiform (gulls and shorebirds) and Passeriform (sparrows and starling). Lately swans have been found infected with H5N1 in a number of European countries (e.g. Austria, Germany, France and Romania, etc.). In poultry, both aquatic and terrestrial species become infected but the virus is particularly aggressive in chicken.

Carnivores: can become infected, after consuming infected poultry that succumbed to the disease. To date no H5N1 clinical cases of dogs have been reported but in an unpublished study carried out in 2005 by the National Institute of Animal Health in Bangkok, researchers tested 629 village dogs and 111 cats in the Suphan Buri district of central Thailand. Out of these, 160 dogs and 8 cats had antibodies to H5N1, indicating that they were infected with the virus or had been infected in the past. An eqiune virus has recently shown up in dogs. This inter-species re-assortment is not uncommon for type A influenza viruses.

Pigs are known “„mixing vessels” for different influenza virus subtypes and therefore present a risk for avian influenza virus re-asserting with a human influenza virus into a strain more apt to infect humans. Regarding the present H5N1 subtype, studies conducted in pigs in Vietnam yielded 8 animals out of the 3000 investigated pigs seropositive. None of the animals had any clinical signs and it was not possible to isolate any virus

Ruminants appear at lower risk. So far no cattle have been identified as carrying any influenza type A virus. Horses are susceptible to Influenza viruses but so far mainly H3N8 have been identified. Regular vaccination is carried out. Experimentally mice can be infected but their role in natural transmission has not been established.

Public health implications
Humans and other mammals need to come in contact with large amounts of virus to become infected. In case of an infection with H5N1, mammals and humans apparently only shed small amounts of virus, contributing to reduced risk of spread among themselves. Recent data from experimentally infected cats’ evidenced extra-respiratory replication of the H5N1 and excretion of virus in faeces of cats need to be taken into consideration. Hygienic practices need to be re-enforced, frequent washing of hands with water and soap especially after handling animals, cleaning cat litter boxes as well as before and after the preparation of food.

Occupational health and safety
Veterinarians and their staff are specifically at risk of coming into contact with infected cats, in case the disease becomes more widespread among this species. Normally, veterinarians and their staff engage in frequent hand washing and disinfect examination tables and instruments to reduce the general risk of disease transmission among their patients and to protect the persons present in the consultation room from eventual exposure.

Advice for veterinarians
The following is advised for veterinarians:

Advice to pet owners (see above)
Be ware of possibility to receive (sick) cats infected with H5N1
Take hygienic measures when handling sick cats (gloves and surgical masks)
Take deep oro-pharyngeal swabs of suspected animals (e.g. animals with respiratory problems) and sent them to the laboratory clearly indicating the type of examination requested Support cases to be reported to veterinary authorities
Inform owners of suspected animals and provide them with clear and practical information, avoiding creating any panic among cat owners or the general public.
Provide veterinarians are advised to contact the Veterinary Authorities in their respective countries for specific instructions

 
 
H5N1 Canine Sequence Similar Feline

Recombinomics Commentary
May 23, 2006

The sequence of canine H5N1, A/
dog/Thailand/Suphanburi/KU-08/04(H5N1). has been released at GenBank. The 2004 isolate is from Thailand and was initially reported in 2004. However, the report was quickly followed by a statement that the H5N1 positive sample was mislabeled. The May 5, 2006 deposit of the dog sequence confirms H5N1 presence in dogs and extends the number of confirmed mammalian species positive for H5N1. The sequence is similar to 2004 domestic, A/cat/Thailand/KU-02/04(H5N1), and wild cat, A/tiger/Thailand/CU-T7/2004(H5N1) isolates from Thailand, including PB2 E627K, a polymorphism that increases virulence in mice. H5N1 bird flu with PB2 E627K can also cause hind leg paralysis in experimental ferrets and has infected civet cats

H5N1 has also been isolated from swine, A/
swine/Anhui/2004(H5N1) and there have been reports of a positive stone martin, providing additional evidence for an expanding mammalian host range.

Recently
H3N8 has also been isolated from dogs in the United States suggesting another mammalian mixing vessel for H5N1 recombination and reassortment..

Media Link

Map

http://www.recombinomics.com/News/05...N1_Canine.html

 
J Virol. 1993 Apr;67(4):1761-4. Related Articles, < =1.2> < =1.2> Links
http://www.ncbi.nlm.nih.gov/entrez/utils/lofref.fcgiPrId=3494&uid=8445709&db=PubMed&url=http://www.pubmedcentral.gov/articlerender.fcgitool=pubmed&pubmedid=8445709 
A single amino acid in the PB2 gene of influenza A virus is a determinant of host range.
http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=8445709&dopt=Citation


Subbarao EK, London W, Murphy BR.

Respiratory Viruses Section, National Institute of Allergy and Infectious Diseases, Bethesda, Maryland 20892.

The single gene reassortant virus that derives its PB2 gene from the avian influenza A/Mallard/NY/78 virus and remaining genes from the human influenza A/Los Angeles/2/87 virus exhibits a host range restriction (hr) phenotype characterized by efficient replication in avian tissue and failure to produce plaques in mammalian Madin-Darby canine kidney cells.

The hr phenotype is associated with restriction of viral replication in the respiratory tract of squirrel monkeys and humans. To identify the genetic basis of the hr phenotype, we isolated four phenotypic hr mutant viruses that acquired the ability to replicate efficiently in mammalian tissue. Segregational analysis indicated that the loss of the hr phenotype was due to a mutation in the PB2 gene itself.

The nucleotide sequences of the PB2 gene of each of the four hr mutants revealed that a single amino acid substitution at position 627 (Glu-->Lys) was responsible for the restoration of the ability of the PB2 single gene reassortant to replicate in Madin-Darby canine kidney cells.

Interestingly, the amino acid at position 627 in every avian influenza A virus PB2 protein analyzed to date is glutamic acid, and in every human influenza A virus PB2 protein, it is lysine.

Thus, the amino acid at residue 627 of PB2 is an important determinant of host range of influenza A viruses.
 
PMID: 8445709 [PubMed - indexed for MEDLINE]

PB2 amino acid at position 627 affects replicative efficiency of Hong Kong H5N1 influenza A viruses in mice.
http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15016548

Shinya K, Hamm S, Hatta M, Ito H, Ito T, Kawaoka Y.

Department of Pathobiological Sciences, University of Wisconsin, Madison, WI 53706, USA.

A single amino acid substitution, from glutamic acid to lysine at position 627 of the PB2 protein, converts a nonlethal H5N1 influenza A virus isolated from a human to a lethal virus in mice. In contrast to the nonlethal virus, which replicates only in respiratory organs, the lethal isolate replicates in a variety of organs, producing systemic infection.
 
Despite a clear difference in virulence and organ tropism between the two viruses, it remains unknown whether the dissimilarity is a result of differences in cell tropism or the reduced replicative ability of the nonlethal virus in mouse cells in general. To determine how this single amino acid change affects virulence and organ tropism in mice, we investigated the growth kinetics of the two H5N1 viruses both in vitro and in vivo. The identity of the PB2 amino acid at position 627 did not appreciably affect viral replicative efficiency in chicken embryo fibroblasts and a quail cell line; however, viruses with lysine at this position instead of glutamic acid grew better in the different mouse cells tested. When the effect of this substitution was investigated in mice, all of the test viruses showed the same cell tropism, but infection by viruses containing lysine at position 627 spread more rapidly than those viruses containing glutamic acid at this position.
 
Further analysis showed a difference in local immune responses: neutrophil infiltration in lungs infected with viruses containing lysine at position 627 persisted longer than that associated with viruses lacking a glutamic acid substitution.
 
Our data indicate that the amino acid at position 627 of the PB2 protein determines the efficiency of viral replication in mouse (not avian) cells, but not tropism among cells in different mouse organs. The presence of lysine leads to more aggressive viral replication, overwhelming the host's defense mechanisms and resulting in high mortality rates in mice.

PMID: 15016548 [PubMed - indexed for MEDLINE]"
 
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http://en.wikipedia.org/wiki/Image:Flu_und_legende_color_c.jpg

"Influenza virus infections in mammals.

Vahlenkamp TW, Harder TC.

Friedrich-Loeffler-Institut, Bundesforschungsinstitut fur Tiergesundheit, Institut fur Molekularbiologie, Greifswald-Insel Riems, Germany. thomas.vahlenkamp@fli.bund.de

The natural reservoir of all known subtypes of influenza A viruses are aquatic birds, mainly of the orders Anseriformes and Charadriiformes in which the infection is asymptomatic and the viruses stay at an evolutionary equilibrium.
 
However, mammals may occasionally contract influenza A virus infections from this pool.
 
This article summarizes: (i) natural infections in mammals including pigs, horses, marine mammals, ferrets, minks; (ii) results from experimental infections in several animal models including mice, ferrets, primates, rats, minks, hamsters and (iii) evidence for the increased pathogenicity of the current influenza A H5N1/Asia viruses for mammals.
 
Several reports have shown that a number of mammalian species, including pigs, cats, ferrets, minks, whales, seals and finally also man are susceptible to natural infection with influenza A viruses of purely avian genetic make up.
 
Among the mammalian species naturally susceptible to avian influenza virus the pig and the cat might exert the greatest potential public health impact.
 
Despite numerous studies in animal and cell culture models, the basis of the extended host spectrum and the unusual pathogenicity of the influenza A H5N1 viruses for mammals is only beginning to be unraveled. Recently, also the transmission of equine influenza A virus to greyhound racing dogs has been documented.

PMID: 16573202 [PubMed - in process] "

Avian influenza (H5N1) viruses isolated from humans in Asia in 2004 exhibit increased virulence in mammals.
http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16140756&query_hl=17&itool=pubmed_docsum

The origins of new pandemic viruses: the acquisition of new host ranges by canine parvovirus and influenza A viruses.
http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16153179&query_hl=30&itool=pubmed_docsum

Evolution In Action: Why Some Viruses Jump Species
http://www.sciencedaily.com/releases/2006/03/060316091731.htm

H5N1 in Dogs and Cats ~"The expanded geographical reach and host range of H5N1 is cause for concern.  The spread of H5N1 allows for more dual infections, recombinations, and new sequences causing new problems.  Moreover, these new sequences can increase the affinity for human receptors, such as S227N, and can be generated in avian hosts infected with H5N1 and H9N2 as predicted previously."
http://www.recombinomics.com/News/02140601/H5N1_Dogs_Cats.html

H5N1 Canine Deaths in Thailand "However, if it is passed from dog to dog, it is another cause for concern because the H5N1 circulating in Thailand generates a very high case fatality rate in humans and there is little difference between H5N1 isolated from birds and cats and H5N1 isolated from fatal human cases." 
http://www.recombinomics.com/news/10040401/canine_deaths.html

H5N1 Bird Flu Infection in Mink Expands Host Range ~ "E627K is found in all human H1, H2, H3 serotypes and is associated with the ability of the polymerase to efficiently function at lower temperatures (33 C) compared to E627 (41 C).  Prior to last year, E627K had not been found in any bird H5N1 isolates.  However, at Qinghai Lake, all 16 bird isolates has E627K and all Qinghai H5N1 strain isolates reported since last year have had E627K. The presence of E627K in all Qinghai isolates may increase the likelihood of mammalian infections from eating H5N1 infected birds.  Therefore, the number of mammalian species infected with H5N1 may be significantly higher than those reported above.  Transmission of H5N1 to other mammalian species increases the likelihood of recombination and acquisition of mammalian polymorphisms, which can lead to an expanded host range. The fixing of E627K in H5N1 in long range migratory birds may have significant impact"
http://www.recombinomics.com/News/03280601/H5N1_Mink.html

H5N1 Bird Flu Detected in Live Cats in Austria
http://www.recombinomics.com/News/03060604/H5N1_Austria_Cats.html

 
Nucleotide banner
 
< = value=98418142 name=uid>1:  DQ530170Reports  Influenza A Virus H5N1...[gi:98418142] Links
LOCUS       DQ530170                2287 bp    RNA     linear   VRL 22-MAY-2006
DEFINITION  Influenza A Virus (A/dog/Thailand-Suphanburi/KU-08/04(H5N1))
            polymerase basic protein 2 gene, complete cds.
ACCESSION   DQ530170
VERSION     DQ530170.1  GI:98418142
KEYWORDS    .
SOURCE      Influenza A Virus (A/dog/Thailand-Suphanburi/KU-08/04(H5N1))
  ORGANISM  Influenza A Virus (A/dog/Thailand-Suphanburi/KU-08/04(H5N1))
            Viruses; ssRNA negative-strand viruses; Orthomyxoviridae;
            Influenzavirus A.
REFERENCE   1  (bases 1 to 2287)
  AUTHORS   Songserm,T., Amonsin,A., Jam-on,R., Sae-Heng,N., Meemak,N.,
            Pariyothorn,N., Damrongwattanapokin,S., Chutinimitkul,S.,
            Payungporn,S., Theamboonlers,A. and Poovorawan,Y.
  TITLE     Direct Submission
  JOURNAL   Submitted (03-MAY-2006) Pediatric, Chulalongkorn University, Sor
            Kor Bld., Pratumwan, Bangkok, Bangkok 10330, Thailand
FEATURES             Location/Qualifiers
     source          1..2287
                     /organism="Influenza A Virus
                     (A/dog/Thailand-Suphanburi/KU-08/04(H5N1))"
                     /mol_type="genomic RNA"
                     /strain="A/Dog/Thailand-Suphanburi/KU-08/04"
                     /serotype="H5N1"
                     /db_xref="taxon:384064"
                     /country="Thailand"
     CDS             1..2280
                     /note="PB2"
                     /codon_start=1
                     /product="polymerase basic protein 2"
                     /protein_id="ABF58844.1"
                     /db_xref="GI:98418143"
                     /translation="MERIKELRDLMSQSRTREILTKTTVDHMAIIKKYTSGRQEKNPA
                     LRMKWMMAMKYPITADKRIIEMIPERNEQGQTLWSKTNDAGSDRVMVSPLAVTWWNRN
                     GPATSAVHYPKVYKTYFEKVERLKHGTFGPVHFRNQVKIRRRVDINPGHADLSAKEAQ
                     DVIMEVVFPNEVGARILTSESQLTITKEKKEELQDCKIAPLMVAYMLERELVRKTRFL
                     PVAGGTSSVYIEVLHLTQGTCWEQMYTPGGEVRNDDVDQSLIIAARNIVRRATVSADP
                     LASLLEMCHSTQIGGIRMVDILRQNPTEEQAVDICKAAMGLRISSSFSFGGFTFKRTS
                     GSSVKKEEEVLTGNLQTLKIRVHEGYEEFTMVGRRATAILRKATRRLIQLIVSGRDEQ
                     SIAEAIIVAMVFSQEDCMIKAVRGDLNFVNRANQRLNPMHQLLRHFQKDAKVLFQNWG
                     IEPIDNVMGMIGILPDMTPSTEMSLRGVRVSKMGVDEYSSTERVVVSIDRFLRVRDQR
                     GNVLLSPEEVSETQGTEKLTITYSSSMMWEINGPESVLVNTYQWIIRNWETVKIQWSQ
                     DPTMLYNKMEFEPFQSLVPKAARGQYSGFVRTLFQQMRDVLGTFDTVQIIKLLPFAAA
                     PPKQSRMQFSSLTVNVRGSGMRILVRGNSPVFNYNKATKRLTVLGKDAGALTEDPDEG
                     TAGVESAVLRGFLILGKEDKRYGPALSINELSNLAKGEKANVLIGQGDVVLVMKRKRD
                     SSILTDSQTATKRIRMAIN"
ORIGIN      
        1 atggagagaa taaaagaatt acgagatcta atgtcacagt cccgcactcg cgagatacta
       61 acaaaaacca ctgtggacca tatggccata atcaagaaat acacatcagg aagacaagag
      121 aagaaccctg ctctcagaat gaaatggatg atggcaatga aatatccaat cacagcggac
      181 aagagaataa tagagatgat tcctgaaagg aatgaacaag ggcagacgct ctggagcaag
      241 acaaatgatg ctggatcgga cagggtgatg gtgtctcccc tagctgtaac ttggtggaat
      301 aggaatgggc cggcgacaag tgcagtccat tatccaaagg tttacaaaac atactttgag
      361 aaggttgaaa ggttaaaaca tggaaccttt ggtcccgttc atttccgaaa ccaagttaaa
      421 atacgccgcc gagttgatat aaatcctggc catgcagatc tcagtgctaa agaagcacaa
      481 gatgtcatca tggaggtcgt tttcccaaat gaagtgggag ccagaatatt gacatcagag
      541 tcgcaattga caataacgaa agagaagaaa gaagagctcc aagattgtaa gattgctccc
      601 ttaatggttg catacatgtt ggaaagggaa ctagtccgca aaaccagatt cctaccggta
      661 gcaggcggaa caagcagtgt gtacattgag gtattgcatt tgactcaagg gacctgctgg
      721 gaacagatgt acactccagg cggagaagtg agaaatgacg atgttgacca gagtttgatc
      781 atcgctgcca gaaacattgt taggagagca acggtatcag cggatccact ggcatcactg
      841 ctggagatgt gtcacagcac acaaattggt gggataagga tggtggacat ccttaggcaa
      901 aatccaactg aggaacaagc tgtggatata tgcaaagcag caatgggtct gaggatcagt
      961 tcttccttta gctttggagg cttcactttc aaaagaacaa gtggatcatc cgtcaagaag
     1021 gaagaggaag tgcttacagg caacctccaa acattgaaaa taagagtaca tgaggggtat
     1081 gaggaattca caatggttgg gcggagggca acagctatcc tgaggaaagc aactagaagg
     1141 ctgattcagt tgatagtaag tggaagagac gaacaatcaa tcgctgaggc aatcattgta
     1201 gcaatggtgt tctcacagga ggattgcatg ataaaggcag tccgaggcga tctgaatttc
     1261 gtaaacagag caaaccaaag attaaacccc atgcatcaac tcctgagaca ttttcaaaag
     1321 gatgcaaaag tgctatttca gaattgggga attgaaccca ttgataatgt catggggatg
     1381 atcggaatat tacctgacat gactcccagc acagaaatgt cactgagagg agtaagagtt
     1441 agtaaaatgg gagtggatga atattccagc actgagagag tagttgtaag tattgaccgt
     1501 ttcttaaggg ttcgagatca gcgggggaac gtactcttgt ctcccgaaga ggtcagcgaa
     1561 acccagggaa cagagaaatt gacaataaca tattcatcat caatgatgtg ggaaatcaac
     1621 ggtcctgagt cagtgcttgt taacacttat cagtggatca tcagaaactg ggagactgtg
     1681 aagattcaat ggtctcaaga ccccacgatg ctgtacaata agatggagtt tgaaccgttc
     1741 caatccttgg tacccaaggc tgccagaggt caatacagtg gatttgtgag aacattattc
     1801 caacaaatgc gtgacgtact ggggacattt gatactgtcc agataataaa gctgctacca
     1861 tttgcagcag ccccaccgaa gcagagcaga atgcagtttt cttctttaac tgtgaatgtg
     1921 agaggctcag gaatgagaat actcgtaagg ggcaattccc ctgtattcaa ctataataag
     1981 gcaaccaaaa ggcttaccgt tcttggaaag gacgcaggtg cattaacaga ggatccagat
     2041 gaggggacag ccggagtgga atctgcagta ctgaggggat tcttaattct aggcaaggag
     2101 gacaaaaggt atgggccagc attgagcatc aatgaactga gcaatcttgc gaagggggag
     2161 aaagctaatg tgctgatagg gcaaggagac gtggtgttgg taatgaaacg gaaacgggac
     2221 tctagcatac ttactgacag ccagacagcg accaaaagaa ttcggatggc catcaattag
     2281 ttgaacg
//
 

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