Flagella and Pili of Xanthomonas axonopodis pv. glycines are associated with motility,biofilm formation and virulence on soybean

Targeted mutations in flgK, and pilD genes in strain KU‐P‐SW005 of Xanthomonas axonopodis pv. glycines, the cause of pustule disease on soybean, led to altered motility phenotypes. The flgK mutants lacked a monopolar flagellum and lost swimming motility, whereas the pilD mutant lacked type IV pili and was unable to move via twitching, a form of surface motility not previously reported for this pathogen. The flgK and pilD mutants were also altered in biofilm production. The flgK and pilD mutants caused reduced disease in susceptible soybean cultivars Spencer when compared to KU‐P‐SW005. Cell counts of the flgK and pilD mutants on plants remained equivalent to KU‐P‐SW005 10 days after inoculation. Complementation of flgK and pilD mutants restored all phenotypes to wild‐type levels. Therefore, flgK and pilD genes that are required for swimming and twitching motility also affected biofilm formation and virulence on soybean.     

Metabarcoding reveals diet diversity in an ungulate community in Thailand

The diverse large mammal communities found in Asian dry forests and savannas should segregate based on their diet selection. We examined the diet composition of sympatric ungulate species using metabarcoding to determine whether their diet was segregated and whether obvious attributes (i.e., body size, phylogeny, ecology) explained the structure. We collected fecal samples from eight ungulate species in Huai Kha Khaeng Wildlife Sanctuary in the western forest complex of Thailand. The fecal collections occurred around a plot where all woody species were codified within a genetic barcode library, and this library was supplemented with samples from plant species known to be consumed by these species. Of 273 plant species tested, at least 93 were found within the fecal samples. Over half of the identified species were not previously known by experts as forage species. All ungulate species showed a strong consumption of grasses and forbs. For the three species with sufficient sample size (sambar, banteng, and guar), there were seasonal differences in their diet, with each showing increased occurrence of woody plants during the dry season. The pattern of forage consumption did not follow obvious paradigms of body size or taxonomy, with significant diet differences found in two similar‐sized bovids (gaur, banteng), while the diet of sambar was more similar to bovids than to the other deer species. Asian ungulates differ in their forage consumption and metabarcoding should allow for testing of diet shifts in response to seasonal rains and fires which dominate the phenology of Asian dry forests and savannas.     

Species composition and vegetation structure of an upper montane forest at the summit of Mt. Doi Inthanon, Thailand

Upper montane forest (UMF) within Doi Inthanon National Park, Northern Thailand, was investigated by means of fifty, 40 × 40 m stratified random plots situated between 2080 and 2565 m altitude. The aim was to address a number of community ecological questions concerning woody species composition and structural heterogeneity of the forest. A total of 7474 individuals of trees and woody climbers ≥ 15 cm gbh (girth at breast height) were included in the study and these were identified to 47 species, 39 genera and 26 families. The average density was 934 individuals/ha and the average stem basal area was 71.8 m²/ha. The most important species were: Quercus eumorpha, Sjzygium angkae, Litsea martabanica, Helicia nilagirica, Lindera caudata, Schima wallichii, Osmanthus fiagrans, Eurya acuminata, Myrsine semiserrata and Ilex umbellulata and the most important families were Fagaceae, Lauraceae, Theaceae and Myrtaceae. Altitude was the most important environmental variable explaining species composition and vegetation structure. Most of the calculated vegetation variables showed significant correlation with altitude: species richness, family richness, diversity, density and crown cover declined with altitude, average tree height was uncorrelated with altitude and basal area increased with altitude. An analysis of size class distributions indicated good forest conditions and reverse-J-shaped age class distribution of most species.     

Influence of fruit on habitat selection of Asian bears in a tropical forest

Wild bear populations in Southeast Asia are threatened with extinction, but the ecology and distribution of the 2 species occurring in the region's protected areas is poorly known, so there is little scientific basis underlying conservation strategies. We used bear signs, primarily claw marks on climbed trees, to study habitat selection and distribution of Asiatic black bear (Ursus thibetanus) and sun bear (Helarctos malayanus) across Khao Yai National Park, Thailand from March to December 2008. We found black bear claw marks in 24 of 30 random sample blocks (80%), indicating that this species was widely distributed across Khao Yai. Sun bear signs were much scarcer: their claw marks occurred in 11 blocks (37%); data were too sparse for sun bear so we limited our focus to Asiatic black bear. Using logistic regression, we found that fruit abundance best explained variation in presence of black bear, whereas human disturbance, distance to park edge, elevation, and ground cover had little influence. Fruits appear to be a key resource for Asiatic black bears, and factors affecting fruit abundance or shifts in seasonality (e.g., climate change) will impact bear populations. Knowledge of this relationship will allow managers to be more proactive in managing bears. We recommend using sign surveys for monitoring changes in black bear occupancy as they are inexpensive, efficient, and can be conducted by trained park rangers. © 2011 The Wildlife Society.     

Arsenic exposure levels of petrochemical workers in three workplace settings in Rayong Province,Thailand

Background: Crude oil and natural gas are often contaminated with arsenic. As a carcinogen, arsenic contamination in the workplace is of concern, particularly when urinary arsenic levels are higher than the standard. The aim of this study was to identify exposure sources of arsenic among petrochemical workers. Methods: A total of 188 operators and 30 office workers participated in the study. Ninety-three workplace air samples, three main meals in five consecutive days, and drinking water were collected from each participant. Urine was collected at the end of the day after the last food sample was collected from each subject. Urine samples where arsenic concentration exceeded 100 mg/L were further analyzed to identify species. Results: The average arsenic concentrations in operators' and office workers' food and urine were 0.55 ± 1.00 and 0.49 ± 0.67 mg/kg; and 76.43 ± 107.36 and 149.92 ± 200.28 mg/L, respectively. The arsenic concentrations in air and water were well below their standards. The urinary arsenic correlated well with arsenic in the food but not in the air and water. Conclusion: Occupational exposure to arsenic among operators and office workers was lower than 1% TLV (Threshold limit value) and did not differ significantly. The major source of arsenic exposure Q2 was food.     

Ecological and Morphological Profile of Floating Spherical Cladophora socialis Aggregations in Central Thailand

The unique beauty of spherical aggregation forming algae has attracted much attention from both the scientific and lay communities. Several aegagropilous seaweeds have been identified to date, including the plants of genus Cladophora and Chaetomorpha. However, this phenomenon remains poorly understood. In July 2013, a mass occurrence of spherical Cladophora aggregations was observed in a salt field reservoir in Central Thailand. The aims of the present study were to describe the habitat of the spherical aggregations and confirm the species. We performed a field survey, internal and external morphological observations, pyrenoid ultrastructure observations, and molecular sequence analysis. Floating spherical Cladophora aggregations (1–8 cm in diameter) were observed in an area ~560 m², on the downwind side of the reservoir where there was water movement. Individual filaments in the aggregations were entangled in each other; consequently, branches growing in different directions were observed within a clump. We suggest that water movement and morphological characteristics promote the formation of spherical aggregations in this species. The molecular sequencing results revealed that the study species was highly homologous to both C. socialis and C. coelothrix. However, the diameter of the apical cells in the study species was less than that of C. coelothrix. The pyrenoid ultrastructure was more consistent with that of C. socialis. We conclude that the study species is C. socialis. This first record of spherical aggregations in this species advances our understanding of these formations. However, further detailed physical measurements are required to fully elucidate the mechanism behind these spherical formations.     

Identification of an operon, Pil-Chp, that controls twitching motility and virulence in Xylella fastidiosa

Xylella fastidiosa is an important phytopathogenic bacterium that causes many serious plant diseases, including Pierce's disease of grapevines. Disease manifestation by X. fastidiosa is associated with the expression of several factors, including the type IV pili that are required for twitching motility. We provide evidence that an operon, named Pil-Chp, with genes homologous to those found in chemotaxis systems, regulates twitching motility. Transposon insertion into the pilL gene of the operon resulted in loss of twitching motility (pilL is homologous to cheA genes encoding kinases). The X. fastidiosa mutant maintained the type IV pili, indicating that the disrupted pilL or downstream operon genes are involved in pili function, and not biogenesis. The mutated X. fastidiosa produced less biofilm than wild-type cells, indicating that the operon contributes to biofilm formation. Finally, in planta the mutant produced delayed and less severe disease, indicating that the Pil-Chp operon contributes to the virulence of X. fastidiosa, presumably through its role in twitching motility.     

Characterization of the Xylella fastidiosa PD1671 Gene Encoding Degenerate c-di-GMP GGDEF/EAL Domains,and Its Role in the Development of Pierce’s Disease

Xylella fastidiosa is an important phytopathogenic bacterium that causes many serious plant diseases including Pierce’s disease of grapevines. X. fastidiosa is thought to induce disease by colonizing and clogging xylem vessels through the formation of cell aggregates and bacterial biofilms. Here we examine the role in X. fastidiosa virulence of an uncharacterized gene, PD1671, annotated as a two-component response regulator with potential GGDEF and EAL domains. GGDEF domains are found in c-di-GMP diguanylate cyclases while EAL domains are found in phosphodiesterases, and these domains are for c-di-GMP production and turnover, respectively. Functional analysis of the PD1671 gene revealed that it affected multiple X. fastidiosa virulence-related phenotypes. A Tn5 PD1671 mutant had a hypervirulent phenotype in grapevines presumably due to enhanced expression of gum genes leading to increased exopolysaccharide levels that resulted in elevated biofilm formation. Interestingly, the PD1671 mutant also had decreased motility in vitro but did not show a reduced distribution in grapevines following inoculation. Given these responses, the putative PD1671 protein may be a negative regulator of X. fastidiosa virulence.     

Occurrence of Three Felids across a Network of Protected Areas in Thailand: Prey,Intraguild, and Habitat Associations

Clouded Leopard, Leopard, and Tiger are threatened felids in Southeast Asia, but little is known about the factors influencing their distributions. Using logistic regression, we assessed how habitat variables, prey detection patterns, and presence of intraguild predators affect the occurrence of these felids across 13 protected areas within Thailand. Our analysis is based on data from 1108 camera‐trap locations (47,613 trap‐nights). Clouded Leopard and Leopard are associated with habitat where Red Muntjac and Eurasian Wild Pig were most likely to be present. Tiger are associated with habitat with a higher likelihood for the presence of Gaur, Eurasian Wild Pig, and Sambar. Clouded Leopard and Tiger were both weakly associated with areas with mature evergreen forest. Besides availability of prey, associations with potential competitors also appear to influence the distribution of these felids, although the strength of these effects requires further investigation. Occurrence rates for Clouded Leopard were no different in protected areas with Leopard versus without Leopards. Leopard had similar occurrence rates regardless of the presence of Tiger, but Leopards were less likely to be detected at the same camera‐trap points with the larger felid. Our results suggest that the two most commonly photographed prey species in the study areas serve as key prey species, Eurasian Wild Pig for all three carnivores and Red Muntjac for Leopard and Clouded Leopard.