Social network analysis and whole-genome sequencing to evaluate disease transmission in a large, dynamic population: A study of avian mycobacteriosis in zoo birds.

This study combined a social network analysis and whole-genome sequencing (WGS) to test for general patterns of contagious spread of a mycobacterial infection for which pathways of disease acquisition are not well understood. Our population included 275 cases diagnosed with avian mycobacteriosis that were nested in a source population of 16,430 birds at San Diego Zoo Wildlife Alliance facilities from 1992 through mid-2014. Mycobacteria species were determined using conventional methods and whole genome sequencing (WGS). Mycobacterium avium avium (MAA) and Mycobacterium genavense were the most common species of mycobacteria identified and were present in different proportions across bird taxa. A social network for the birds was constructed from the source population to identify directly and indirectly connected cases during time periods relevant to disease transmission. Associations between network connectivity and genetic similarity of mycobacteria (as determined by clusters of genotypes separated by few single nucleotide polymorphisms, or SNPs) were then evaluated in observed and randomly generated network permutations. Findings showed that some genotypes clustered along pathways of bird connectivity, while others were dispersed throughout the network. The proportion of directly connected birds having a similar mycobacterial genotype was 0.36 and significant (p<0.05). This proportion was higher (0.58) and significant for MAA but not for M. genavense. Evaluations of SNP distributions also showed genotypes of MAA were more related in connected birds than expected by chance; however, no significant patterns of genetic relatedness were identified for M. genavense, although data were sparse. Integrating the WGS analysis of mycobacteria with a social network analysis of their host birds revealed significant genetic clustering along pathways of connectivity, namely for MAA. These findings are consistent with a contagious process occurring in some, but not all, case clusters.

Bird flu, influenza and 1918: the case for mutant Avian tuberculosis.

Influenza is Italian for "influence", Latin: influentia. It used to be thought that the disease was caused by a bad influence from the heavens. Influenza was called a virus long, long before it was proven to be one. In 2005, an article in the New England Journal of Medicine estimated that a recurrence of the 1918 influenza epidemic could kill between 180 million and 360 million people worldwide. A large part of the current bird-flu hysteria is fostered by a distrust among the lay and scientific community regarding the actual state of our knowledge regarding the bird flu or H5N1 and the killer "Influenza" Pandemic of 1918 that it is compared to. And this distrust is not completely unfounded. Traditionally, "flu" does not kill. Experts, including Peter Palese of the Mount School of Medicine in Manhattan, remind us that even in 1992, millions in China already had antibodies to H5N1, meaning that they had contracted it and that their immune system had little trouble fending it off. Dr. Andrew Noymer and Michel Garenne, UC Berkely demographers, reported in 2000 convincing statistics showing that undetected tuberculosis may have been the real killer in the 1918 flu epidemic. Aware of recent attempts to isolate the "Influenza virus" on human cadavers and their specimens, Noymer and Garenne summed that: "Frustratingly, these findings have not answered the question why the 1918 virus was so virulent, nor do they offer an explanation for the unusual age profile of deaths". Bird flu would certainly be diagnosed in the hospital today as Acute Respiratory Distress Syndrome (ARDS). Roger and others favor suspecting tuberculosis in all cases of acute respiratory failure of unknown origin. By 1918, it could be said, in so far as tuberculosis was concerned, that the world was a supersaturated sponge ready to ignite and that among its most vulnerable parts was the very Midwest where the 1918 unknown pandemic began. It is theorized that the lethal pig epidemic that began in Kansas just prior to the first human outbreaks was a disease of avian and human tuberculosis genetically combined through mycobacteriophage interchange, with the pig, susceptible to both, as its involuntary living culture medium. What are the implications of mistaking a virus such as Influenza A for what mycobacterial disease is actually causing? They would be disastrous, with useless treatment and preventative stockpiles. The obvious need for further investigation is presently imminent and pressing.

Polymerase chain reaction identification of Mycobacterium avium in formalin-fixed, paraffin-embedded animal tissues.

A PCR procedure previously developed for identification of Mycobacterium bovis in formalin-fixed tissues was used to identify mycobacteria of the M. avium complex. Tissues were examined from 100 culture-positive cases of M. avium complex infection, including 86 in which the subspecies was not identified and 14 that had been identified as M. avium subsp. paratuberculosis. Each sample was tested with 5 primer sets, 16S ribosomal RNA (rRNA), IS900, IS901, IS1245, and a heat shock protein (hspX), that detect 1 or both M. avium subspecies. The success rate of PCR detection varied with the primers used and the animal species tested. Among the 86 cases with no M. avium subspecies designation, primers for the 16S rRNA gene were clearly the most efficient because they produced amplicons from all samples that reacted with any other primer set. The overall detection rate in this group of samples was 71%: highest in avian tissues (89%) followed by swine (72%) and ruminants (57%) None of the avian or swine tissues reacted with primers for IS900 or hspX, which identify M. a. paratuberculosis. In contrast, 7 of the 12 ruminant samples that were 16S rRNA positive reacted with 1 or both of these primers. All of the 14 cases shown by culture to be M. a. paratuberculosis infections were positive with IS900 primers, whereas only 11 were positive for 16S rRNA. These results indicate that 16S rRNA primers are the most useful for PCR identification of M. avium in formalin-fixed tissues of nonruminant species. However, IS900 primers should also be used when ruminant tissues are examined because these primers provide the greatest sensitivity for detection of M. a. paratuberculosis infections.

Susceptibility of captive wildfowl to avian tuberculosis: the importance of genetic and environmental factors.

This study reports the findings of an epidemiological survey of death due to avian tuberculosis in the captive collection of wildfowl at The Wildfowl and Wetlands Trust Centre, Slimbridge, Gloucestershire. Both genetic and environmental factors have been shown to affect the incidence of, and the birds' susceptibility to, the disease. Seasonal body condition was related to the occurrence of death due to the disease in both males and females. Birds from either hot or cold climates appeared to have a higher incidence than those from temperate climates. What the birds ate did not affect incidence but the method they used for obtaining their food did. Higher susceptibility was found in those species evolved for marine or arboreal habitats. Anomalies in susceptibility which suggest a higher level of genetic immunity in some groups have also been found. Reasons are put forward to explain these findings.