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Applying the scientific method when assessing the influence of migratory birds on the dispersal of H5N1

Sally Furniss

Well-known member
Applying the scientific method when assessing the influence of migratory birds on the dispersal of H5N1

Paul L Flint
U.S. Geological Survey, Alaska Science Center, 1011 East Tudor Road, Anchorage, Alaska, 99503, USA

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Virology Journal 2007, 4:132doi:10.1186/1743-422X-4-132
The electronic version of this article is the complete one and can be found online at: http://www.virologyj.com/content/4/1/132
<table cellpadding="0" cellspacing="0"> <tbody> <tr> <td>Received:</td> <td>21 November 2007</td> </tr> <tr> <td>Accepted:</td> <td>4 December 2007</td> </tr> <tr> <td>Published:</td> <td>4 December 2007</td> </tr> </tbody> </table> ? 2007 Flint; licensee BioMed Central Ltd.
This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Abstract

Background

The role of wild birds in the dispersal of highly pathogenic avian influenza virus H5N1 continues to be the subject of considerable debate. However, some researchers functionally examining the same question are applying opposing null hypotheses when examining this issue.
Discussion

I describe the correct method for establishing a null hypothesis under the scientific method. I suggest that the correct null hypothesis is that migratory birds can disperse this virus during migration and encourage researchers to design studies to falsify this null. Finally, I provide several examples where statements made during this debate, while strictly true, are not generally informative or are speculative.
Summary

By adhering to the scientific method, definitive answers regarding the role of wild birds in the dispersal of highly pathogenic viruses will be reached more effectively.
Background

Considerable debate remains regarding the role of wild birds in the dispersal of the highly pathogenic avian influenza (HPAI) H5N1 virus. Numerous articles have been published on this topic, many of which lack any data which would allow critical testing relevant to this issue. Throughout the literature, there are two opposing views with regard to the assumptions regarding to the role of wild birds in dispersing H5N1. First, some authors assume that wild birds cannot disperse this virus over long distances and cite the lack of studies demonstrating such movements [1-4]. Conversely, other authors presume that wild birds can disperse this virus [5-8]. These two opposing views are based on reversal of the functional null hypothesis. The goal of this paper is to apply the scientific method to the development of the appropriate null hypothesis for this issue.
Discussion

The most appropriate null hypothesis

The implied null hypotheses are Ho(1): Birds can disperse H5N1 during migration; and Ho(2): Birds cannot disperse H5N1 during migration. Both are potentially valid null hypotheses and each uses the other as the alternative. Thus the main question becomes, which is more appropriate as a working null hypothesis? The development of a null hypothesis, under the scientific method, is predicated on the desire to control the probability of making a type I error. Recall that type I errors occur when a null hypothesis is rejected when in fact the null hypothesis is true [9]. Thus, choosing between these two potential null hypotheses requires assessing which of the alternative type I errors is more severe. Type I error from Ho(1): Conclude that migratory birds cannot disperse H5N1 when in fact they do; Type I error from Ho(2): Conclude that migratory birds do disperse H5N1 when in fact they do not. Because there are currently no data available that would allow a critical test of either of these hypotheses we are left with considering which of these type I errors is most severe or unacceptable. Since the primary interest in H5N1 is relative to the potential risk to the poultry industry and human health, it would seem that prematurely dismissing a potential carrier for dispersal has the potential to allow this virus to expand its range undetected as well as hinder the response to detected outbreaks. Conversely, incorrectly concluding that migratory birds can disperse this virus is simply inefficient. That is, time and resources may be wasted sampling wild birds to detect virus dispersal that is not occurring. However, this inefficiency is only relevant to human health risks if associated resources would be re-directed to issues associated with alternative pathways of H5N1 dispersal (i.e., by poultry transport). Given the funding processes associated with these programs, such re-allocation would seem unlikely. Accordingly, Ho(1) is the appropriate working null hypothesis as the type I error based on this null hypothesis is the most severe and the probability of making this error should be minimized and controlled. While acknowledging that under the strict scientific method, a null hypothesis is never accepted as being true (that is, null hypotheses can only be falsified), it is common practice, particularly in the field of wildlife biology where researchers have little control over potential covariates, to functionally assume the working null as being true while actively attempting to falsify the null. Researchers should pursue data which will allow a critical test of the null hypothesis that wild birds can disperse H5N1 during migration; particularly long distance migration. Importantly, sufficient resources need to be allocated to this effort such that failure to reject this null is associated with reasonable statistical power (i.e., minimizing the probability of a type II error where we fail to reject a null that is false)
Uninformative conclusions drawn from valid results

Finally, throughout the debate on the respective roles of migratory and domestic birds in the dispersal of H5N1, there are numerous statements that are not based on data presented, or when they are based on data, are not necessarily conclusive. Two such examples are detailed below. First, it has been concluded that wild birds are not an important carrier because HPAI viruses such as H5N1 are rarely isolated from apparently healthy wild birds [10]. However, the same statement can be made about domestic chickens in areas where H5N1 is considered endemic [5]. Thus, the same logic, as applied to migratory birds, could be used to conclude that domestic poultry are not an important carrier. In a second example, it has been speculated that wild birds could not effectively transport H5N1 over long distances as migratory performance would be negatively influenced by the infection [4]. While others pointed out that H5N1 appeared to kill wild birds nearly as efficiently as domestic poultry and noted that "dead ducks don't fly" [1]. However, many of the same arguments would likely apply to long distance transport of domestic poultry. During transport, domestic birds are frequently deprived of food and water for extended periods and are exposed to extreme environmental conditions. Again, the same logic could be used to conclude that exposed domestic poultry are less likely to survive long distance transport for the same immunological reasons. In both of these examples, while the original statements may be strictly true, the conclusions drawn from them are not necessarily valid or informative.
Conclusion

As the debate regarding the role of wild birds and domestic fowl in the dispersal of H5N1 continues, researchers should be careful to draw conclusions directly from data (i.e., avoid speculation) and ensure that conclusions are empirically and logically supported by all available data. By following the scientific principles, collecting data required for critical tests of hypotheses, and avoiding speculation, definitive conclusions regarding the dispersal of H5N1 will be reached more effectively.



Abbreviations

HPAI: highly pathogenic avian influenza
Competing interests

The author(s) declare that they have no competing interests.
Acknowledgements

The author thanks D. Derksen, D. Rocque, J. Schmutz and H. Wilson for critical evaluation of the manuscript. The author was supported by the U.S. Geological Survey during development of this manuscript.
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http://www.virologyj.com/content/4/1/132
 
Re: Applying the scientific method when assessing the influence of migratory birds on the dispersal of H5N1

There is no scientific debate. There was no reported H5N1 west of China prior to Qinghai Lake. After, ALL H5N1 was the Qinghai strain. The "debate" ended in the summer of 2005. False negatives using a fatally flawed assay do not provide statistical power.
 
Re: Applying the scientific method when assessing the influence of migratory birds on the dispersal of H5N1

you see, what strange things occur from the trend that scientists
don't give numbers to quantify their claims - here the amount of
spread of H5N1 over long distances by wild birds.

The question is not "if" or "if not" but "how much".
I guesstimate, that 70% of introductions to new locations
are due to wild birds.
 
Re: Applying the scientific method when assessing the influence of migratory birds on the dispersal of H5N1

I guesstimate, that 70% of introductions to new locations
are due to wild birds.
Interesting but then the question is what figures do you use to arrive at 70% and where do the other 30% come from?
 
Re: Applying the scientific method when assessing the influence of migratory birds on the dispersal of H5N1

With a premise that articles on
Virology Journal 2007, 4:132doi:10.1186/1743-422X-4-132
are enaugh scientific for the debate,
we can see that
the statistical minimum of data needed to support, or not, scientificaly one of the debata side, are not reached yet:

"Because there are currently no data available that would allow a critical test of either of these hypotheses we are left with considering which of these type I errors is most severe or unacceptable."

Second, from the article text,
the side stated that wild birds are the long distance carriers must:

"Researchers should pursue data which will allow a critical test of the null hypothesis that wild birds can disperse H5N1 during migration; particularly long distance migration."

It means that, the researchers, before stating that the wild birds are the main spreaders, must previously pursued the data which corroborate that statement, not the ones saying the birds are not the main spreaders.

Third, from the conclusion text of the article:

"... researchers should be careful to draw conclusions directly from data (i.e., avoid speculation) and ensure that conclusions are empirically and logically supported by all available data."

From the article on virology, the conclusions of the researchers cannot be drawed only from genetic data, but must be supported by fisical evidence (empiricaly) in the field, and supported by all other kind of available data, not only genetical (logically).
 
Re: Applying the scientific method when assessing the influence of migratory birds on the dispersal of H5N1

Interesting but then the question is what figures do you use to arrive at 70% and where do the other 30% come from?

mainly what I read in the forums, the papers, the outbreaks.
And my experience with relationship of sequences.

The 30% come from the difference to 100% : 30%=100%-70%.
No "proof" yet, so it can't be 100%.

Well, I could list some pros and cons ...
Biggest con: very little H5N1 found in testing healthy wild birds.

Hey, we could make a poll : how much of the H5N1-spread do you
estimate is due to birds ?
But few votes on flutrackers usually, so I also it on another forum.
 
Re: Applying the scientific method when assessing the influence of migratory birds on the dispersal of H5N1

With a premise that articles on
Virology Journal 2007, 4:132doi:10.1186/1743-422X-4-132
are enaugh scientific for the debate,
we can see that
the statistical minimum of data needed to support, or not, scientificaly one of the debata side, are not reached yet:

"Because there are currently no data available that would allow a critical test of either of these hypotheses we are left with considering which of these type I errors is most severe or unacceptable."

Second, from the article text,
the side stated that wild birds are the long distance carriers must:

"Researchers should pursue data which will allow a critical test of the null hypothesis that wild birds can disperse H5N1 during migration; particularly long distance migration."

It means that, the researchers, before stating that the wild birds are the main spreaders, must previously pursued the data which corroborate that statement, not the ones saying the birds are not the main spreaders.

Third, from the conclusion text of the article:

"... researchers should be careful to draw conclusions directly from data (i.e., avoid speculation) and ensure that conclusions are empirically and logically supported by all available data."

From the article on virology, the conclusions of the researchers cannot be drawed only from genetic data, but must be supported by fisical evidence (empiricaly) in the field, and supported by all other kind of available data, not only genetical (logically).
Papers that start with an assumption that there is a debate are just looking to use negative data for more hocus pocus. These papers are along the lines of intelligent design - no science required.
 
Re: Applying the scientific method when assessing the influence of migratory birds on the dispersal of H5N1

The existence of a debate cannot reasonably be denied by refering to
(debating) conflicting papers.

I think there is pretty much debate about this.
More than I had expected.
 
Re: Applying the scientific method when assessing the influence of migratory birds on the dispersal of H5N1

Papers that start with an assumption that there is a debate are just looking to use negative data for more hocus pocus. These papers are along the lines of intelligent design - no science required.

All scientists are driven by apriori assumptions and biased world views. There is no such thing as unbiased research. A true scientist acknowledges his/her assumptions and world view upfront, and attempts to ensure that they do not bias their interpretation of research.
 
Last edited by a moderator:
Re: Applying the scientific method when assessing the influence of migratory birds on the dispersal of H5N1

The existence of a debate cannot reasonably be denied by refering to
(debating) conflicting papers.

I think there is pretty much debate about this.
More than I had expected.
The "debates" are started by conservation groups. There is no debate on the science side. The data could not be clearer.
 
Re: Applying the scientific method when assessing the influence of migratory birds on the dispersal of H5N1

All scientific theories which claim to be "ultimate" must be proved by fisical evidence.
If it is realy:
"There is no debate on the science side. The data could not be clearer."

That exactly means in terms of evidence what:
The main world spreaders of H5N1 are the wild birds, many of them are resistant to the virus, asymptomatic, and can shed it over long distances?

I can than continue with:
We can do nothing to stop that, but we can try to isolate the animals mainly deadly affected by the virus - the poultry.

Or the treatment will be worst than the illness - kill all the wild organisms? I hope it will not. That kind of radical measures remains doubtful for the already high infected zones also (range 1,5 miles).


If somebody have better suggestions for tretments, wrote them.
 
Re: Applying the scientific method when assessing the influence of migratory birds on the dispersal of H5N1

All scientific theories which claim to be "ultimate" must be proved by fisical evidence.
If it is realy:
"There is no debate on the science side. The data could not be clearer."

That exactly means in terms of evidence what:
The main world spreaders of H5N1 are the wild birds, many of them are resistant to the virus, asymptomatic, and can shed it over long distances?

I can than continue with:
We can do nothing to stop that, but we can try to isolate the animals mainly deadly affected by the virus - the poultry.

Or the treatment will be worst than the illness - kill all the wild organisms? I hope it will not. That kind of radical measures remains doubtful for the already high infected zones also (range 1,5 miles).


If somebody have better suggestions for tretments, wrote them.
The debate ended in the summer of 2005. The Qinghai strain was identified in wild birds in May, 2005 at Qinghai Lake. Conservation groups used "dead birds don't fly" and "wild birds as victims" arguments to say the strain would burn out at Qinghai Lake and were infected because of domestic poultry. In July, the outbreak at Chany Lake in Russia ended the debate. The wild birds were thousands of miles from Qinghai Lake (they can fly), and a healthy teal was infected with the Qinghai strain, Russia had never reported H5N1 previously, The outbreak in Russia was followed by Erhel Lake in remote Monogolia. Same story. No domestic poultry around, wild birds infected with Qinghai strain. Mongolia had never reported H5N1 previosu. Same story repeat 50X in the next year in countries west of China. None had reported H5N1 previously. ALL reported Qinghai. MANY had ZERO reports in domestic poultry.

The case was closed long ago.

Conservation groups use an assay that doesn't work *they found ZERO H5N1 positive live birds at Erhel Lake, where there was clearly H5N1 in the dead birds (and live birds that were missed by the assay).

The data really doesn't get any clearer.

The "debates" are simply propaganda exercises by conservation groups.

All of the above was known TWO YEARS ago.
 
Re: Applying the scientific method when assessing the influence of migratory birds on the dispersal of H5N1

mainly what I read in the forums, the papers, the outbreaks.
And my experience with relationship of sequences.

The 30% come from the difference to 100% : 30%=100%-70%.
No "proof" yet, so it can't be 100%.

Well, I could list some pros and cons ...
Biggest con: very little H5N1 found in testing healthy wild birds.

Hey, we could make a poll : how much of the H5N1-spread do you
estimate is due to birds ?
But few votes on flutrackers usually, so I also it on another forum.
Please no more hocus pocus.

Bernard Mathews was the exception that proved the rule. H5N1 in their turkeys in England were 99.94% identical with ducks in Hungary.

Please name ONE example of outbreaks on farms that are more than 100 miles apart and have H5N1 with a similar level of identity.

Name ONE H5N1 from a farm, wild bird, or person west of China that is NOT clade 2.2 (Qinghai strain).

30% is pure fantasy - with ZERO data.
 
Re: Applying the scientific method when assessing the influence of migratory birds on the dispersal of H5N1

I came across this study by Victor Nettles, John Wood and Robert Webster dated January of 1985; I didn't realize this debate had been going on for almost 25 years.

Here, when they tested for H5N2 antibodies were detected in 33% of the waterfowl. Doesn't our much more updated testing in wild birds at least show antibodies against H5N1?

http://links.jstor.org/sici?sici=0005-2086(198507/09)29:3<733:WSAWAO>2.0.CO;2-R#abstract
Wildlife surveillance was conducted for influenza viruses in conjunction with the 1983-84 lethal H5N2 avian influenza epizootic in domestic poultry in Pennsylvania, New Jersey, Maryland, and Virginia. Virus-isolation attempts made on cloacal and tracheal swabs from 4,466 birds and small rodents within the quarantined areas and 1,511 waterfowl in nearby Maryland yielded only a single H5N2 isolate from a pen-raised chukar in Pennsylvania. Antibodies against hemagglutinin type 5 and/or neuraminidase type 2 were found in 33% of the aquatic birds tested; however, this finding could not be used to confirm previous H5N2 avian influenza virus activity because of the possibility of prior infections with multiple influenza subtypes. The low prevalence of lethal H5N2 avian influenza virus in wild birds and small rodents strongly indicated that these animals were not responsible for dissemination of the disease among poultry farms during the outbreak.
 
Re: Applying the scientific method when assessing the influence of migratory birds on the dispersal of H5N1

The debate ended in the summer of 2005. The Qinghai strain was identified in wild birds in May, 2005 at Qinghai Lake.
...
The case was closed long ago.
...
... there was clearly H5N1 in the dead birds (and live birds that were missed by the assay).
...
The data really doesn't get any clearer.
...
All of the above was known TWO YEARS ago.

Thank you Dr. Niman for your answer.

I agree that's the case for the past debate in 2005.,
but after that we must continue with the previously wroted situation:

"The main world spreaders of H5N1 are the wild birds, many of them are resistant to the virus, asymptomatic, and can shed it over long distances".

My comments wasn't on the 2005. situation, but currently.

The debate like I saw it, is which part of the spreading chain can be broken, to assure the illness to not spread on much more teritory.

For now, seems that the countries without endemic bird flu doing well with the (prev. wroted) policy:
"We can do nothing to stop that (the wild birds infections), but we can try to isolate the animals mainly deadly affected by the virus - the poultry."

Is that the reason why after the 2005. the high virulent bf version is not spread more through the poultry flocks, or not?

If not, we can suppose that the role of the wild birds in spreading are overstimated.

If better ways of breaking the infection chain by other ways exists, I hope that they will be described here at FT also.
 
Re: Applying the scientific method when assessing the influence of migratory birds on the dispersal of H5N1

what antibodies do they test for ?
Isn't it that they just pick some epitopes, amino-acid chains of lengths
about 20, and then check whether they appear in/at the antibodies ?
So they test for H5 and then for N2, but not H5N2 ? Could be
H7N2 reassorted or H5N1.
The "and/or" is strange. What do they mean ?
So it should be possible to test for Qinghai-H5-antibodies ?!?
(but expensive ?)

I think, they tested for antibodies in HK when there was an outbreak,
don't remember the result. H5-antibodies in wild birds should be rare
and if, then it's presumably low-path.



I came across this study by Victor Nettles, John Wood and Robert Webster dated January of 1985; I didn't realize this debate had been going on for almost 25 years.

Here, when they tested for H5N2 antibodies were detected in 33% of the waterfowl. Doesn't our much more updated testing in wild birds at least show antibodies against H5N1?

http://links.jstor.org/sici?sici=0005-2086(198507/09)29:3<733:WSAWAO>2.0.CO;2-R#abstract
Wildlife surveillance was conducted for influenza viruses in conjunction with the 1983-84 lethal H5N2 avian influenza epizootic in domestic poultry in Pennsylvania, New Jersey, Maryland, and Virginia. Virus-isolation attempts made on cloacal and tracheal swabs from 4,466 birds and small rodents within the quarantined areas and 1,511 waterfowl in nearby Maryland yielded only a single H5N2 isolate from a pen-raised chukar in Pennsylvania. Antibodies against hemagglutinin type 5 and/or neuraminidase type 2 were found in 33% of the aquatic birds tested; however, this finding could not be used to confirm previous H5N2 avian influenza virus activity because of the possibility of prior infections with multiple influenza subtypes. The low prevalence of lethal H5N2 avian influenza virus in wild birds and small rodents strongly indicated that these animals were not responsible for dissemination of the disease among poultry farms during the outbreak.
 
Re: Applying the scientific method when assessing the influence of migratory birds on the dispersal of H5N1

I came across this study by Victor Nettles, John Wood and Robert Webster dated January of 1985; I didn't realize this debate had been going on for almost 25 years.

Here, when they tested for H5N2 antibodies were detected in 33% of the waterfowl. Doesn't our much more updated testing in wild birds at least show antibodies against H5N1?

http://links.jstor.org/sici?sici=0005-2086(198507/09)29:3<733:WSAWAO>2.0.CO;2-R#abstract
Wildlife surveillance was conducted for influenza viruses in conjunction with the 1983-84 lethal H5N2 avian influenza epizootic in domestic poultry in Pennsylvania, New Jersey, Maryland, and Virginia. Virus-isolation attempts made on cloacal and tracheal swabs from 4,466 birds and small rodents within the quarantined areas and 1,511 waterfowl in nearby Maryland yielded only a single H5N2 isolate from a pen-raised chukar in Pennsylvania. Antibodies against hemagglutinin type 5 and/or neuraminidase type 2 were found in 33% of the aquatic birds tested; however, this finding could not be used to confirm previous H5N2 avian influenza virus activity because of the possibility of prior infections with multiple influenza subtypes. The low prevalence of lethal H5N2 avian influenza virus in wild birds and small rodents strongly indicated that these animals were not responsible for dissemination of the disease among poultry farms during the outbreak.
H5 antibodies in wild birds is common. The conservation groups avoid that area. They just focus on a fataly flawed assay that they use 350,000 times.
 
Re: Applying the scientific method when assessing the influence of migratory birds on the dispersal of H5N1

what antibodies do they test for ?
Isn't it that they just pick some epitopes, amino-acid chains of lengths
about 20, and then check whether they appear in/at the antibodies ?
So they test for H5 and then for N2, but not H5N2 ? Could be
H7N2 reassorted or H5N1.
The "and/or" is strange. What do they mean ?
So it should be possible to test for Qinghai-H5-antibodies ?!?
(but expensive ?)

I think, they tested for antibodies in HK when there was an outbreak,
don't remember the result. H5-antibodies in wild birds should be rare
and if, then it's presumably low-path.
Please. H5 is common in wild birds (and assays can be designed to distiguish between high and low path, just as ferret antibodies can distingusih between Brisbane and Solomon Islands'like seasonal flu).
 
Re: Applying the scientific method when assessing the influence of migratory birds on the dispersal of H5N1

Please no more hocus pocus.

Bernard Mathews was the exception that proved the rule. H5N1 in their turkeys in England were 99.94% identical with ducks in Hungary.

Please name ONE example of outbreaks on farms that are more than 100 miles apart and have H5N1 with a similar level of identity.

Name ONE H5N1 from a farm, wild bird, or person west of China that is NOT clade 2.2 (Qinghai strain).

30% is pure fantasy - with ZERO data.


100 miles is not much, we should test these outbreaks in Korea,Germany,...
but usually the exact location is not given.
Denmark-Scotland is about 500 miles, 4 or 5 nucleotide-differences
in the 3 polymerases.
Italy-Iran : 30 mutations, ~2000km
Lioaning-Shantou : 24 mutations(some segments missing) , ~2000km
I just only checked Qinghai-viruses.
Who finds the record-holder with minimum #mutations/geographic distance ?
(except B.Matthews)
I get about 80km per mutation


I'm not aware of the rule, that exceptions proof the rule.
That "rule" itself would be a paradoxon.
OK, poultry-introduction examples: B.Matthews,Nigeria,Germany 2007 (ducks,
don't remember exactly)...
The current outbreaks in H7 usually spread by poultry, I think. The first introduction
maybe by wild birds, though.


---------------------

edit:
current record : Kurgan-Nigeria, 7000km, 45 mutations in the whole genome
 
Re: Applying the scientific method when assessing the influence of migratory birds on the dispersal of H5N1

100 miles is not much, we should test these outbreaks in Korea,Germany,...
but usually the exact location is not given.
Denmark-Scotland is about 500 miles, 4 or 5 nucleotide-differences
in the 3 polymerases.
Italy-Iran : 30 mutations, ~2000km
Lioaning-Shantou : 24 mutations(some segments missing) , ~2000km
I just only checked Qinghai-viruses.
Who finds the record-holder with minimum #mutations/geographic distance ?
(except B.Matthews)


I'm not aware of the rule, that exceptions proof the rule.
That "rule" itself would be a paradoxon.
OK, poultry-introduction examples: B.Matthews,Nigeria,Germany 2007 (ducks,
don't remember exactly)...
The current outbreaks in H7 usually spread by poultry, I think. The first introduction
maybe by wild birds, though.
H5N1 in Denmark, Scotland, Sweden, and northern Germany in early 2006 all involved wild birds and most of the isolates were the same clade 2.2 subclade, 2.2.2.
 
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