First Report of the Armillaria Root-Disease Pathogen, Armillaria gallica, Associated with Several Woody Hosts in Three States of Mexico.

In September 2007, rhizomorphs with morphological characteristics of Armillaria were collected from woody hosts in forests of Mexico State, Veracruz, and Oaxaca, Mexico. Based on pairing tests, isolates were assigned to five somatically compatible genets or clones (MEX7R, MEX11R, MEX23R, MEX28R, and MEX30R). These genets were all identified as Armillaria gallica based on somatic pairing tests against known tester isolates and nucleotide sequences of the translation elongation factor 1α (tef-1α; GenBank Accession Nos. KF156772 to 76). Sequences of tef-1α for all genets showed a max identity of 97 to 99% to A. gallica (ST23, JF313125) (3,4). However, A. gallica comprises a genetically diverse complex that likely represents multiple cryptic species (3). In Mexico, this species has been previously reported in northeastern Morelos on Quercus sp., eastern Mexico State on Pinus hartwegii, and southwestern Mexico State on Prunus persica (1,2). This study identified associations with 10 new hosts within three states of Mexico, but only five hosts were diseased. Genet MEX7R comprised seven isolates collected in the University of Chapingo forest near Texcoco, Mexico State (19°18'10.764″ N, 98°42'14.147″ W, elevation 3441 m). Four MEX7R isolates were collected from diseased Alnus sp. including the root ball of a 130 cm dbh, root-disease killed tree, one isolate from a symptomless Senecio sp. s.l. (Roldana sp.) shrub and two isolates from symptomless Abies religiosa. Genet MEX11R comprised four isolates from a cloud forest near Xalapa, Veracruz (19°31'14.628″ N, 96°59'22.812″ W, elevation 1496 m). MEX11R isolates were collected from the roots of a root-disease killed Carpinus caroliniana, and from trees with no obvious symptoms (Miconia mexicana, Quercus xalapensis, and Liquidambar styraciflua). Two isolates of genet MEX23R were collected from the Jardin Botanico Francisco Javier Clavijero, Instituto de Ecologia, A.C., Xalapa, Veracruz (19°30'49.067″ N, 96°56'32.999″ W, elevation 1344 m). These isolates were from root-diseased Eriobotrya japonica (non-native fruit tree) that showed obvious symptoms (flaccid, chlorotic, and senescing leaves) and from an adjacent, infected Platanus mexicana that did not show readily observable symptoms. Two collections near Oaxaca, Oaxaca, included a single isolate MEX28R from the root ball of a recently root disease-killed Arbutus xalapensis within a small root disease center at Peña Prieta, in Parque La Cumbre, near Ixtepeji (17°09'42.084″ N, 96°38'15.936″ W, elevation 2853 m) and a single isolate MEX30R from the base of an asymptomatic Alnus acuminata near the El Carrizal fish hatchery 10 km northeast of San Miguel del Valle (17°06'45.036″ N, 96°24'03.743″ W, elevation 2594 m). Armillaria gallica has a circumpolar distribution with an extremely wide host range, and its ecological behavior varies greatly. Continued surveys are needed to better understand the distribution and ecological impacts of this pathogen in relation to Armillaria root disease in Mexico and the potential influences of climate change. Although A. gallica displays diverse ecological behavior, trees infected with A. gallica are less likely to survive the stresses of human activity and a changing climate (4). References: (1) D. Alvarado-Rosales and R. A. Blanchette. Phytopathology 84:1106, 1994. (2) R. D. Elias-Roman et al. For. Pathol. 43:390, 2013. (3) M.-S. Kim et al. Phytopathology 102:S4.63, 2012. (4) B. Marcais and N. Breda. J. Ecol. 94:1214, 2006.

First Report of the Armillaria Root Disease Pathogen, Armillaria gallica, on Douglas-fir (Pseudotsuga menziesii) in Arizona.

In August 2010, a mycelial fan (isolate AZ32F) of Armillaria sp. was collected from the root collar of a living Douglas-fir tree on the Mogollon Rim within the Coconino National Forest (approximate location 34°25'31.26″N, 111°20'41.04″W, elevation 2,293 m) in central Arizona. Mycelial fans under the bark of living trees are a sign of pathogenicity, and symptoms of the diseased tree included resinosis, sloughing bark, and thinning crown. The infected tree was located on a south-facing slope with approximately 30% tree cover, dominated by ponderosa pine (Pinus ponderosa), with lesser components of Douglas-fir and Gambel oak (Quercus gambelii). Based on three replications of somatic incompatibility tests against 24 tester isolates representing seven North American Armillaria spp., isolate AZ32F showed 100% intraspecific compatibility (colorless antagonism) with all four A. gallica isolates, 22% compatibility with A. calvescens, and 0% compatibility with the remaining Armillaria spp. Based on GenBank BLASTn of isolate AZ32F sequences, the partial LSU-IGS1 (GenBank Accession No. KF186682) showed 99 to 100% similarity to A. gallica and two other related Armillaria spp. with 99 to 100% coverage, and translation elongation factor-1 alpha (tef-1α) sequences (KC525954) showed 96% similarity to A. gallica (JF895844) with 100% coverage. Thus, isolate AZ32F was identified as A. gallica, based on somatic incompatibility tests and DNA sequences (partial LSU-IGS1 and tef-1α). Although the isolate is identified as A. gallica with similarities to other North American isolates, evidence is mounting that currently recognized A. gallica likely represents a species complex that comprises multiple phylogenetic species (4). Previous surveys in Arizona have noted A. mellea and A. solidipes (as A. ostoyae) (3), but A. gallica has never been previously confirmed in this state. Within North America, A. gallica is commonly reported east of the Rocky Mountains and in West Coast states of the United States, where it infects hardwoods and conifers including Douglas-fir (1,2). Its ecological behavior ranges from saprophyte to weak/aggressive pathogen (1,2). Because damage by A. gallica appears to increase on hosts predisposed by stress (1), further surveys are needed to document its distribution, frequency, and ecological behavior in the southwestern United States, where climate change will likely cause tree stress due to maladaptation. Continued surveys for Armillaria spp. will better determine their potential threat within the geologically and ecologically unique Mogollon Rim of Arizona. References: (1) K. Baumgartner and D. M. Rizzo. Plant Dis. 85:947, 2001. (2) N. J. Brazee and R. L. Wick. For. Ecol. Manage. 258:1605, 2009. (3) R. L. Gilbertson and D. M. Bigelow. J. Arizona-Nevada Acad. Sci. 31:13, 1998. (4) M.-S. Kim et al. Phytopathology 102:S4.63, 2012.

First Report of an Armillaria Root Disease Pathogen, Armillaria gallica, Associated with Several New Hosts in Hawaii.

The loss and decline of native tree species caused by invasive plant pathogens is a major threat to the endangered endemic forests of the Hawaiian Islands (3). Thus, it is critical to characterize existing pathogens to evaluate potential invasiveness. In August 2005, rhizomorphs and mycelial bark fans of genet HI-4 were collected from dead/declining, mature trees of introduced Monterey pine (Pinus radiata) on the southern flank of Mauna Kea, Hawaii (approximately 19°42'55″N, 155°26'48″W, elevation 2,175 m). In March of 2008, three additional genets (HI-11, HI-13, and HI-16) were collected as rhizomorphs at a site named Pu'u La'au (west slope of the Mauna Kea Forest Reserve area, approximately 19°50'00″N, 155°35'35″W, elevation 2,275 to 2,550 m), approximately 20 km west-northwest of the HI-4 collection. These genets were collected from apparently healthy loblolly pine (Pinus taeda) that were introduced, apparently healthy mamane (Sophora chrysophylla; an endemic tree species of Hawaii), dead and dying mamane, and apparently healthy Methley plum (Prunus cerasifera × Prunus salicina) that was planted. All isolates were determined to have identical sequences in the intergenic spacer-1 rDNA region (GenBank Accession No. DQ995357). On the basis of somatic paring tests against North American Armillaria tester strains and 99% nucleotide sequence identities to GenBank Accession Nos. AY190245 and AY190246, these isolates were identified as Armillaria gallica. Past surveys have noted A. mellea sensu lato and A. nabsnona on numerous hosts in Hawaii, including mamane (3,4). However, to our knowledge, this is the first confirmed report of A. gallica in Hawaii, where it was found on mamane, Monterey pine, loblolly pine, and Methley plum. A. gallica has been widely categorized as a beneficial saprophyte, an opportunistic pathogen, or an aggressive pathogen (2). A recent study suggests that A. gallica can be highly pathogenic in some areas of the eastern United States and it is an important component of forest decline (2), especially under increasing stressors such as climate change. The isolation of A. gallica from declining stands on both introduced and endemic hosts under drought conditions suggests this pathogen is a contributing factor to forest decline on the island of Hawaii. Because the mamane tree is an important component of the native forest stands and essential to the endangered palila bird (Loxioides bailleui), which feeds almost exclusively on its green seeds (1), continued monitoring of Armillaria root disease is warranted. References: (1) P. C. Banko et al. J. Chem. Ecol. 28:1393, 2002. (2) N. J. Brazee and R. L. Wick. For. Ecol. Manage. 258:1605, 2009. (3) R. E. Burgan and R. E. Nelson. USDA For. Serv. Tech. Rep. PSW-3, 1972. (4) J. W. Hanna et al. Plant Dis. 91:634, 2007.

First Report of Armillaria Root Disease Caused by Armillaria tabescens on Araucaria araucana in Veracruz, Mexico.

In September 2007, bark samples were collected from the root collar of a single Araucaria araucana tree that had recently died and was suspected of being killed by Armillaria root disease. Disease symptoms and signs included a thinning crown and fruiting bodies at the tree base over a several-year period before tree death. The tree was located in an isolated street-tree planting within a business district on Maestros Veracruzanos Street, Xalapa, Veracruz (19°31'52''N, 96°54'25''W, elevation 1,392 m). One fungal isolate (MEX21WF) was obtained, which possessed two sequence repeat types from the intergenic spacer-1 (IGS-1) region (GenBank Accession Nos. GQ335541 and GQ335542). On the basis of these IGS-1 sequences, this isolate from Mexico possessed 99% nucleotide sequence identities with North American Armillaria tabescens isolates (GenBank Accession Nos. AY695410 ≈ GQ335541 and AY773966 ≈ GQ335542). Somatic pairing tests of the isolate with other North American Armillaria species also identified it as A. tabescens (2). In addition, fruiting bodies were produced on the stump base in 2009 that matched morphological features of A. tabescens, e.g., exannulate, cespitose growth in clusters, brown-gray stipe to blackish toward the base, longitudinally fibrillose, basidiospores (6-) 7 to 9 × 4 to 5 (-5.5) µm, and other general morphology. On the basis of these three lines of taxonomic evidence, it was concluded that the isolate was A. tabescens. To our knowledge, this is the first confirmed report of A. tabescens causing Armillaria root disease in Mexico. Furthermore, this note represents the first report of A. tabescens on Araucaria araucana, which is native to Chile and Argentina. The other previous reports of A. tabescens in Mexico are based on herbarium specimens collected in 1965 (BPI 753040) from Valle de Bravo (approximately 350 km west of Xalapa) in the state of México and 1973 (BPI 753041) from near Monterrey (approximately 760 km north-northwest of Xalapa) in the state of Nuevo León (1). However, no host information or confirmation of taxonomic identification was reported for these herbarium specimens. Although this note confirms the presence of A. tabescens in Mexico, more surveys and monitoring are needed to determine the full distribution of this pathogen in Mexico. Because the climate and tree communities of eastern Mexico are similar to those of the southeastern United States, where A. tabescens has been reported as a common pathogen of oaks and fruit trees (3,4), it seems reasonable that A. tabescens may represent an existing or potential threat in eastern Mexico. References: (1) D. F. Farr and A. Y. Rossman. Fungal Databases. Systematic Mycology and Microbiology Laboratory. Online publication. ARS, USDA, 2010. (2) K. I. Mallett and Y. Hiratsuka. Can. J. Bot. 64:2588, 1986. (3) F. Miranda and A. J. Sharp. Ecology 31:313, 1950. (4) G. Schnabel et al. Mycol. Res. 109:1208, 2005.

First Report of Armillaria sinapina, a Cause of Armillaria Root Disease, Associated with a Variety of Forest Tree Hosts on Sites with Diverse Climates in Alaska.

In August of 2007, a preliminary survey was conducted in Alaska to evaluate potential impacts of climate change on forest trees. Armillaria sinapina, a root-disease pathogen, was isolated from conifer and hardwood hosts on climatically diverse sites spanning 675 km from the Kenai Peninsula to the Arctic Circle. Seven isolates (NKAK1, NKAK2, NKAK5, NKAK6, NKAK9F, NKAK13, and NKAK15) were identified as A. sinapina by using intergenic spacer-1 nucleotide sequences (GenBank Accession Nos. EU665175-EU665181) and somatic pairings. Of particular note is that one isolate (NKAK9F) was obtained from a declining Salix sp. (willow) growing in a flood plain near the Arctic Circle (66°32.316'N, 150°47.717'W). This isolate was collected from mycelial bark fans that were intercalated within multiple bark layers, a sign of disease. All other isolates were derived from rhizomorphs attached to and/or embedded within roots and root collars, but most host trees showed no clear indication of disease. Two isolates were collected from dead trees within a small mortality center (62°08.703'N, 150°04.593'W) that included an isolate from Picea glauca (white spruce; NKAK13) and another isolate from Betula sp. (birch; NKAK15). Additional isolates came from a beetle-killed P. glauca (NKAK1) 120 km northwest of Anchorage (61°48.079'N, 148°16.983'W) and a suppressed (overtopped by other trees in the stand) Tsuga mertensiana (mountain hemlock; NKAK2) 58 km southeast of Anchorage (60°50.679'N, 149°03.742'W). The two remaining isolates originated from the Kenai Peninsula (approximately 60°29.629'N, 149°45.465'W) and were derived from a root-diseased Populus tremuloides (trembling aspen; NKAK5) and a suppressed P. glauca (NKAK6). Although A. mellea sensu lato was previously reported on willow in interior Alaska (1) and A. sinapina was previously reported from sites under coastal influence (4), this represents the first confirmed report of A. sinapina on P. glauca, T. mertensiana, Populus tremuloides, Salix sp., and Betula sp. in Alaska. Unfortunately, pathogenicity of A. sinapina cannot be readily verified under experimental conditions because environmental variables, host-tree status (e.g., species, population, age, and vigor), and inoculum potential are difficult to recreate. Armillaria sinapina is typically regarded as a weak pathogen of diverse hosts (3). However, A. sinapina is predicted to cause more disease on hosts predisposed by climate stress, and climate change is well-documented in Alaska (2). Because A. sinapina occurs on diverse hosts under different climates across a wide geographic range in Alaska, Armillaria root disease could become more prevalent on trees stressed by climate change. References: (1) T. E. Hinds and T. H. Laurent. Plant Dis. Rep. 62:972, 1978. (2) J. J. McCarthy et al., eds. Climate Change 2001: Impacts, Adaptation and Vulnerability. Cambridge University Press, Cambridge, 2001. (3) D. J. Morrison et al. Can. J. Plant Pathol. 7:242, 1985. (4) C. G. Shaw, III and E. M. Loopstra. Phytopathology 78:9714, 1988.

First Report of the Root-Rot Pathogen, Armillaria nabsnona, from Hawaii.

The genus Armillaria (2) and Armillaria mellea sensu lato (3) have been reported previously from Hawaii. However, Armillaria species in Hawaii have not been previously identified by DNA sequences, compatibility tests, or other methods that distinguish currently recognized taxa. In August 2005, Armillaria rhizomorphs and mycelial bark fans were collected from two locations on the island of Hawaii. Stands in which isolates were collected showed moderate to heavy tree mortality and mycelial bark fans. Pairing tests (4) to determine vegetative compatibility groups revealed three Armillaria genets (HI-1, HI-7, and HI-9). Rhizomorphs of genet HI-1 were collected from both dead and healthy mature trees of the native 'Ohia Lehua (Metrosideros polymorpha) approximately 27 km west of Hilo, HI (approximately 19°40'49″N, 155°19'24″W, elevation 1,450 m). Rhizomorphs of HI-7 and HI-9 were collected, respectively, from dead/declining, mature, introduced Nepalese alder (Alnus nepalensis) and from an apparently healthy, mature, introduced Chinese banyan (Ficus microcarpa) in the Waipi'o Valley (approximately 20°03'29″N, 155°37'35″W, elevation 925 m). On the basis of somatic pairing tests and intergenic spacer-1 (IGS-1) nucleotide sequence identities of 99 to 100% with North American A. nabsnona (GenBank Accession No. AY509178), HI-1 (GenBank Accession No. DQ995356), HI-7 (GenBank Accession No. DQ995358), and HI-9 (GenBank Accession No. DQ995359) were identified as A. nabsnona, a pathogen of hardwoods (1). The IGS-1 sequences of A. nabsnona genets (HI-1, HI-7, and HI-9) had a greater similarity to North American collections of A. nabsnona than to the Asian A. nabsnona, even though the two introduced hosts originated from Asia. Phylogeographic studies could help determine the potential introduction and original source of A. nabsnona in Hawaii. Although A. nabsona was isolated from multiple hosts in declining stands, pathogenicity studies are needed to confirm whether this pathogen causes disease on diverse native and exotic tree species in Hawaii. References: (1) E. Allen et al. Pages 2-7 in: Common Tree Diseases of British Columbia. Natural Resources Canada. Canadian Forest Service, Victoria, BC, Canada, 1998. (2) D. E. Hemmes and D. E. Desjardin. Pages 129 and 153 in: Mushrooms of Hawaii. Ten Speed Press, Berkeley, CA, 2002. (3) F. F. Laemmlen and R. V. Bega. Plant Dis. Rep. 58:102, 1974. (4) Y. Wu et al. USDA Forest Service Tech. Rep. R2-58, 1996.

Pleural infections: a clinical-radiologic review.

Pleural space infection is a common disorder that may result from a wide variety of causes and is associated with a wide range of etiologic agents. The authors reviewed retrospectively records of 102 patients with discharge diagnoses of empyema and/or bronchopleural fistula. Chest radiographs and computed tomography closest to the time of initial diagnosis were evaluated separately. In 78 cases of empyema, the etiologies included primary pulmonary infections (49%), postsurgical (23%), traumatic (11.5%), intraabdominal pathology (5%), and unknown (11.5%). In 24 cases of bronchopleural fistulas, the etiologies were previous surgical procedures (37.5%), pulmonary infections (37.5%), traumatic (4%), and unknown (21%).