Heinrich zu Dohna

Emergence and Genetic Variation of Neuraminidase Stalk Deletions in Avian Influenza Viruses

When avian influenza viruses (AIVs) are transmitted from their reservoir hosts (wild waterfowl and shorebirds) to domestic bird species, they undergo genetic changes that have been linked to higher virulence and broader host range. Common genetic AIV modifications in viral proteins of poultry isolates are deletions in the stalk region of the neuraminidase (NA) and additions of glycosylation sites on the hemagglutinin (HA). Even though these NA deletion mutations occur in several AIV subtypes, they have not been analyzed comprehensively. In this study, 4,920 NA nucleotide sequences, 5,596 HA nucleotide and 4,702 HA amino acid sequences were analyzed to elucidate the widespread emergence of NA stalk deletions in gallinaceous hosts, the genetic polymorphism of the deletion patterns and association between the stalk deletions in NA and amino acid variants in HA. Forty-seven different NA stalk deletion patterns were identified in six NA subtypes, N1–N3 and N5–N7. An analysis that controlled for phylogenetic dependence due to shared ancestry showed that NA stalk deletions are statistically correlated with gallinaceous hosts and certain amino acid features on the HA protein. Those HA features included five glycosylation sites, one insertion and one deletion. The correlations between NA stalk deletions and HA features are HA-NA-subtype-specific. Our results demonstrate that stalk deletions in the NA proteins of AIV are relatively common. Understanding the NA stalk deletion and related HA features may be important for vaccine and drug development and could be useful in establishing effective early detection and warning systems for the poultry industry.

Invasions by Eurasian avian influenza virus H6 genes and replacement of the virus' North American clade.

This study showed frequent cross-hemisphere virus movement, which can affect the risk posed to poultry and humans.

Spatial re-establishment dynamics of local populations of vectors of Chagas disease.

Background Prevention of Chagas disease depends mainly on control of the insect vectors that transmit infection. Unfortunately, the vectors have been resurgent in some areas. It is important to understand the dynamics of reinfestation where it occurs. Here we show how continuous- and discrete-time models fitted to patch-level infestation states can elucidate different aspects of re-establishment. Triatoma infestans, the main vector of Chagas disease, reinfested sites in three villages in northwest Argentina after community-wide insecticide spraying in October 1992. Methodology/Principal Findings Different methods of estimating the probabilities of bug establishment on each site were compared. The results confirmed previous results showing a 6-month time lag between detection of a new infestation and dispersal events. The analysis showed that more new bug populations become established from May to November than from November to May. This seasonal increase in bug establishment coincides with a seasonal increase in dispersal distance. In the fitted models, the probability of new bug establishment increases with increasing time since last detected infestation. Conclusions/Significance These effects of season and previous infestation on bug establishment challenge our current understanding of T. infestans ecology and highlight important gaps in knowledge. Experiments necessary to close these gaps are discussed.

Adaptation and transmission of a duck-origin avian influenza virus in poultry species

A duck-origin avian influenza virus (AIV) was used to study viral adaptation and transmission patterns in chickens (Gallus gallus domesticus) and Pekin ducks (Anas platyrhynchos domesticus). Inoculated birds were housed with naïve birds of the same species and all birds were monitored for infection. The inoculating duck virus was transmitted effectively by contact in both species. Viruses recovered from infected birds showed mutations as early as 1 or 3 days after inoculation in chickens and ducks, respectively. Amino acid substitutions in hemagglutinin (HA) or deletions in neuraminidase (NA) stalk regions were identified in chicken isolates, but only substitutions in HA were identified in duck isolates. HA substitution-containing viruses replicated more efficiently than those with NA stalk deletions. NA deletion mutants were not recovered from contact chickens, suggesting inefficient transmission. Amino acid substitutions in HA proteins appeared in pairs in chickens, but were independent in ducks, indicating adaptation in chickens. In addition, our findings showed that a duck-origin virus can rapidly adapt to chickens, suggesting that the emergence of new epidemic AIV can be rapid.

Wildlife-livestock interactions in a western rangeland setting: quantifying disease-relevant contacts.

Disease transmission between wild ungulates and domestic livestock is an important and challenging animal health issue. The potential for disease transmission between wildlife and livestock is notoriously difficult to estimate. The first step for estimating the potential for between-species disease transmission is to quantify proximity between individuals of different species in space and time. This study estimates second-order statistics of spatio-temporal location data from radio-collared free-ranging deer, elk and cattle in northeast Oregon. Our results indicate, that when observed simultaneously, elk and cattle occur in closer proximity to each other than what would be expected based on general space use of these species. The same is true for deer and elk but not for deer and cattle. Our analysis also demonstrates that average distances between cattle and elk are largely driven by rare events of close co-mingling between the species, which extend over several hours. Behavioral causes for these co-mingling events are currently unknown. Understanding the causes for such events will be important for designing grazing practices that minimize wildlife-livestock contacts.

Evaluation of the benefit of emergency vaccination in a foot-and-mouth disease free country with low livestock density

Foot-and-mouth disease (FMD) is highly contagious and one of the most economically devastating diseases of cloven-hoofed animals. Scientific-based preparedness about how to best control the disease in a previously FMD-free country is therefore essential for veterinary services. The present study used a spatial, stochastic epidemic simulation model to compare the effectiveness of emergency vaccination with conventional (non-vaccination) control measures in Switzerland, a low-livestock density country. Model results revealed that emergency vaccination with a radius of 3 km or 10 km around infected premises (IP) did not significantly reduce either the cumulative herd incidence or epidemic duration if started in a small epidemic situation where the number of IPs is still low. However, in a situation where the epidemic has become extensive, both the cumulative herd incidence and epidemic duration are reduced significantly if vaccination were implemented with a radius of 10 km around IPs. The effect of different levels of conventional strategy measures was also explored for the non-vaccination strategy. It was found that a lower compliance level of farmers for movement restrictions and delayed culling of IPs significantly increased both the cumulative IP incidence and epidemic duration. Contingency management should therefore focus mainly on improving conventional strategies, by increasing disease awareness and communication with stakeholders and preparedness of culling teams in countries with a livestock structure similar to Switzerland; however, emergency vaccination should be considered if there are reasons to believe that the epidemic may become extensive, such as when disease detection has been delayed and many IPs are discovered at the beginning of the epidemic.

Fitting parameters of stochastic birth-death models to metapopulation data.

Populations that are structured into small local patches are a common feature of ecological and epidemiological systems. Models describing this structure are often referred to as metapopulation models in ecology or household models in epidemiology. Small local populations are subject to demographic stochasticity. Theoretical studies of household disease models without resistant stages (SIS models) have shown that local stochasticity can be ignored for between patch disease transmission if the number of connected patches is large. In that case the distribution of the number of infected individuals per household reaches a stationary distribution described by a birth-death process with a constant immigration term. Here we show how this result, in conjunction with the balancing condition for birth-death processes, provides a framework to estimate demographic parameters from a frequency distribution of local population sizes. The parameter estimation framework is applicable to estimate parameters of disease transmission models as well as metapopulation models.

Identifying errors in avian influenza virus gene sequences and implications for data usage of public databases.

Public gene sequence databases have become important research tools to understand viruses and other organisms. Evidence suggests that the identifying information for some of the sequences in these databases might not belong to the sequences they are associated with. We developed two tests to conduct a comprehensive analysis of all published sequences of the hemaglutinin and neuramidase genes of avian influenza viruses (AIVs) to identify sequences that may have been misclassified. One test identified sequence pairs with highly similar nucleotide sequences despite a difference of several years between their sampling dates. Another test, which was applied to samples sequenced and deposited more than once, detected sequences with more nucleotide differences to their own than to their closest relatives. All sequences identified as misclassified were further traced to relevant publications to assess the likelihood of contamination and determine if any conclusions were associated with the use of these sequences. Our results suggested that among 4040 published gene sequences examined, approximately 0.8% might be misclassified and that publications using these sequences may include inaccurate statements. Findings from this report suggest that using laboratory-adapted strains and handling multiple samples simultaneously increases the risk of contamination. The tests reported here may be useful for screening new submissions to public sequence databases.