Can We Use Functional Annotation of Prokaryotic Taxa (FAPROTAX) to Assign the Ecological Functions of Soil Bacteria?

FAPROTAX is a promising tool for predicting ecological relevant functions of bacterial and archaeal taxa derived from 16S rRNA amplicon sequencing. The database was initially developed to predict the function of marine species using standard microbiological references. This study, however, has attempted to access the application of FAPROTAX in soil environments. We hypothesized that FAPROTAX was compatible with terrestrial ecosystems. The potential use of FAPROTAX to assign ecological functions of soil bacteria was investigated using meta-analysis and our newly designed experiments. Soil samples from two major terrestrial ecosystems, including agricultural land and forest, were collected. Bacterial taxonomy was analyzed using Illumina sequencing of the 16S rRNA gene and ecological functions of the soil bacteria were assigned by FAPROTAX. The presence of all functionally assigned OTUs (Operation Taxonomic Units) in soil were manually checked using peer-reviewed articles as well as standard microbiology books. Overall, we showed that sample source was not a predominant factor that limited the application of FAPROTAX, but poor taxonomic identification was. The proportion of assigned taxa between aquatic and non-aquatic ecosystems was not significantly different (p > 0.05). There were strong and significant correlations (σ = 0.90–0.95, p < 0.01) between the number of OTUs assigned to genus or order level and the number of functionally assigned OTUs. After manual verification, we found that more than 97% of the FAPROTAX assigned OTUs have previously been detected and potentially performed functions in agricultural and forest soils. We further provided information regarding taxa capable of N-fixation, P and K solubilization, which are three main important elements in soil systems and can be integrated with FAPROTAX to increase the proportion of functionally assigned OTUs. Consequently, we concluded that FAPROTAX can be used for a fast-functional screening or grouping of 16S derived bacterial data from terrestrial ecosystems and its performance could be enhanced through improving the taxonomic and functional reference databases

Life on the wall: the diversity and activity of microbes on 13th – century AD. Lan Na mural painting

Diverse microorganisms from the three domains of life (Archaea, Bacteria, and Eukaryota) cause deterioration in mural paintings worldwide; however, few studies have simultaneously targeted these three domains. This study aims to survey the microbiome and its potential for biodeterioration on unpreserved Lan Na mural paintings in Sean Khan temple, Chiang Mai, Thailand. The overview of the archaeal, bacterial, and fungal communities was reported by Illumina sequencing, whereas the potential for deterioration was revealed by culturable techniques and a literature search. The abundant microbes reported in this study were also found in other ancient mural paintings worldwide. Halococcus, a salt-tolerant archaeon, as well as the eubacterial genus Crossiella dominated the prokaryotic community. On the other hand, the main fungal group was the genus Candida (Ascomycota). However, a low number of fungi and bacteria were isolated. Most of the isolates showed the ability to survive in the drought conditions of mural paintings but could not perform discoloration activities. The deterioration activity mainly affected calcium compounds, which are the main components of painting substrates. Aspergillus and several bacterial isolates could dissolve calcium compounds, but only Trichaptum species could induce crystal formation. These results suggest that deterioration of painting substrate should be taken into consideration in addition to deterioration of color in mural paintings. For the Lan Na painting in Sean Khan temple, the plaster is the prime target for biodeterioration, and thus we suggest that the preservation effort should focus on this component of the mural painting

A new species of Lepidotrigona (Hymenoptera: Apidae) from Thailand with the description of males of L. flavibasis and L. doipaensis and comments on asymmetrical genitalia in bees

Attasopa, Korrawat, Bänziger, Hans, Disayathanoowat, Terd, Packer, Laurence (2018): A new species of Lepidotrigona (Hymenoptera: Apidae) from Thailand with the description of males of L. flavibasis and L. doipaensis and comments on asymmetrical genitalia in bees. Zootaxa 4442 (1): 63-82, DOI: 10.11646/zootaxa.4442.1.

Prevalence of infection by the microsporidian Nosema spp. in native bumblebees (Bombus spp.) in northern Thailand.

Bumblebees (tribe Bombini, genus Bombus Latreille) play a pivotal role as pollinators in mountain regions for both native plants and for agricultural systems. In our survey of northern Thailand, four species of bumblebees (Bombus (Megabombus) montivagus Smith, B. (Alpigenobombus) breviceps Smith, B. (Orientalibombus) haemorrhoidalis Smith and B. (Melanobombus) eximius Smith), were present in 11 localities in 4 provinces (Chiang Mai, Mae Hong Son, Chiang Rai and Nan). We collected and screened 280 foraging worker bumblebees for microsporidia (Nosema spp.) and trypanosomes (Crithidia spp.). Our study is the first to demonstrate the parasite infection in bumblebees in northern Thailand. We found N. ceranae in B. montivagus (5.35%), B. haemorrhoidalis (4.76%), and B. breviceps (14.28%) and N. bombi in B. montivagus (14.28%), B. haemorrhoidalis (11.64%), and B. breviceps (28.257%)

Attributes of host-specificity better explain the diversified wood-boring longhorn beetles in tropical SW China than plant species diversity

Abstract A long-debated question in ecology is whether the hyper-diversity of tropical plant-feeding insects is a direct consequence of high tropical plant diversity or should be attributed to increases in host plant specialization. In this study, we used Cerambycidae (the wood-boring longhorn beetles whose larval stages feed on the xylems of trees and lianas) and plants as study materials to explore which hypothesis is more favoured. Multiple analyses were used to show the differences in host specificity of Cerambycidae in tropical and subtropical forests. From these analyses, we found that the alpha diversity of beetles in tropical forests was significantly higher than that in subtropical forests but not in plants. The relationship between plants and beetles was also closer in tropical areas than in subtropical areas. Our results imply that the wood-boring longhorn beetles show higher degrees of niche conservatism and host-specificity in tropical forests than in subtropical forests. The high diversity of wood-boring longhorn beetles in tropical forests might be explained to a large extent by their more finely partitioned diet breadth

Anti-Herpes Simplex Virus and Anti-Inflammatory Activities of the Melittin Peptides Derived from <i>Apis mellifera</i> and <i>Apis florea</i> Venom

Herpes simplex virus (HSV) is known to cause cold sores and various diseases in humans. Importantly, HSV infection can develop latent and recurrent infections, and it is also known to cause inflammation. These infections are difficult to control, and effective treatment of the disease remains a challenge. Thus, the search for new antiviral and anti-inflammatory agents is a necessity. Melittin is a major peptide that is present in the venom of the honeybee. It possesses a number of pharmacological properties. In this study, the effects of the melittin peptides from A. mellifera (MEL-AM) and A. florea (MEL-AF) against HSV-1 and HSV-2 were evaluated at different stages during the viral multiplication cycle in an attempt to define the mode of antiviral action using plaque reduction and virucidal assays. The results revealed a new finding that melittin at 5 µg/mL demonstrated the highest inhibitory effect on HSV through the direct inactivation of viral particles, and MEL-AF displayed a greater virucidal activity. Moreover, melittin was also observed to interfere with the process of HSV attachment to the host cells. MEL-AM exhibited anti-HSV-1 and anti-HSV-2 effects with EC50 values of 4.90 ± 0.15 and 4.39 ± 0.20 µg/mL, while MEL-AF demonstrated EC50 values of 4.47 ± 0.21 and 3.95 ± 0.61 µg/mL against HSV-1 and HSV-2, respectively. However, non-cytotoxic concentrations of both types of melittin produced only slight degrees of HSV-1 and HSV-2 inhibition after viral attachment, but melittin at 5 µg/mL was able to reduce the plaque size of HSV-2 when compared to the untreated group. In addition, MEL-AM and MEL-AF also exhibited anti-inflammatory activity via the inhibition of nitric oxide production in LPS-stimulated RAW 264.7 macrophage cells, and they were also found to down-regulate the expressions of the iNOS, COX-2 and IL-6 genes. The highest inhibition of IL-6 mRNA expression was found after treatment with 10 µg/mL of MEL-AM and MEL-AF. Therefore, melittin peptides have displayed strong potential to be used as an alternative treatment for HSV infection and inflammatory diseases in the future

Volatile organic compounds of Thai honeys produced from several floral sources by different honey bee species.

The volatile organic compounds (VOCs) of four monofloral and one multifloral of Thai honeys produced by Apis cerana, Apis dorsata and Apis mellifera were analyzed by headspace solid-phase microextraction (HS-SPME) followed by gas chromatography and mass spectrometry (GC-MS). The floral sources were longan, sunflower, coffee, wild flowers (wild) and lychee. Honey originating from longan had more VOCs than all other floral sources. Sunflower honey had the least numbers of VOCs. cis-Linalool oxide, trans-linalool oxide, ho-trienol, and furan-2,5-dicarbaldehyde were present in all the honeys studied, independent of their floral origin. Interestingly, 2-phenylacetaldehyde was detected in all honey sample except longan honey produced by A. cerana. Thirty-two VOCs were identified as possible floral markers. After validating differences in honey volatiles from different floral sources and honeybee species, the results suggest that differences in quality and quantity of honey volatiles are influenced by both floral source and honeybee species. The group of honey volatiles detected from A. cerana was completely different from those of A. mellifera and A. dorsata. VOCs could therefore be applied as chemical markers of honeys and may reflect preferences of shared floral sources amongst different honeybee species

Bacterial communities associated with the ectoparasitic mites Varroa destructor and Tropilaelaps mercedesae of the honey bee (Apis mellifera)

Varroa and Tropilaelaps mites have been reported as serious ectoparasites of the honey bee (Apis mellifera). In this study, bacterial communities associated with Varroa destructor and Tropilaelaps mercedesae from northern Thailand were determined, using both culture-dependent and culture-independent approaches. Adult female mites were collected from apiaries in Chiang Mai and Lampang provinces. Culturable bacteria were isolated from individual mites. On average, we observed approximately 1340 and 1140 CFU/mite in Varroa and Tropilaelaps, respectively. All isolates were assigned to the genus Enterococcus. Six samples of genomic DNA from 30-50 mites were extracted and subjected to pyrosequencing of bacterial 16S rRNA amplicons. The resulting 81 717 sequences obtained from Varroa were grouped into 429 operational taxonomic units. The most abundant bacteria in Varroa mites belonged to the family Enterobacteriaceae, especially the genera Arsenophonus, Enterobacter and Proteus. For Tropilaelaps mites, 84 075 sequences were obtained and clustered into 166 operational taxonomic units, within which the family Enterococcaceae (particularly the genus Enterococcus) was predominant. Localization of bacteria in the mites using fluorescence in situ hybridization with two universal bacterial probes revealed that these bacteria were in the cecum of the mites. Taxon-specific Enterobacteriaceae and Arsenophonus probes also confirmed their localization in the cecum of Varroa

<i>Tropilaelaps mercedesae</i> Infestation Is Correlated with Injury Numbers on the Brood and the Population Size of Honey Bee <i>Apis mellifera</i>

Tropilaelaps mercedesae, one of the most devastating parasitic mites of honey bee Apis mellifera hosts, is a major threat to honey products by causing severe damage to honey bee colonies. Here, we recorded injury numbers caused by T. mercedesae to different body parts of the larval, pupal, and crippled adult stages of honey bee A. mellifera. We evaluated the relationship between infestation rate and injury numbers per bee for both larvae and pupae. We also noted the total bee numbers per beehive and examined the relationship between the infestation rate and population size. T. mercedesae infested all developmental stages of honey bees, with the highest injury numbers in the abdomens of bee pupae and the antennas of crippled adult bees. Although larvae received more injury numbers than pupae, both infestation rate and injury numbers decreased as the larval stage progressed to the pupal stage. The infestation rate increased as the population size per beehive decreased. This study provided new perspectives to the understanding of changes in the effects of T. mercedesae infestations on different developmental stages of honey bees. It also showed useful baseline information for screening honey bee stock that might have high defensive behaviors against mite infestation

Properties of essential oils absorbed on the surface of cardboard pieces after using atmospheric-pressure plasma treatments to develop long-lasting Varroa miticides in honeybees (Apis mellifera).

The ectoparasitic mite, Varroa destructor is the most serious widespread pest of managed honeybees (Apis mellifera). Several acaricide products, which include essential oils, have been proposed for mite control. In this study, we aimed to apply atmospheric-pressure plasma to modify a cardboard piece surface in order to prolong the delivery of essential oils for controlling Varroa in honeybee colonies. Absorption capacity, release rates and evaporation rates of essential oils were determined. Cardboard piece showed a higher absorption capacity of cinnamon compared to citronella and clove. Surface modification of cardboard pieces using argon plasma at different gas flow rates and treatment durations, significantly affected the absorption of clove oil. Additionally, the release rate of cinnamon, citronella and clove was significantly enhanced after argon plasma treatments. Evaporation of cinnamon was dramatically increased by plasma treatment at 6-h of incubation. The highest evaporation rate was obtained by plasma-treated cardboard piece at a gas flow rate of 0.5 Lpm for 60 s (0.2175 ± 0.0148 μl/g•h). Efficiency of plasma-treated cardboard piece, impregnated with essential oils, was also investigated for Varroa control in honeybee colonies. In the first experiment, formic acid 65% (v/v) showed the highest efficiency of 90.60% and 81.59% with the percent of mite infestation was 0.23 ± 0.13% and 0.47 ± 0.19% at 21 and 35 days, respectively after treatment. The efficacy of cardamon oil (5% (v/v)) delivered using plasma-treated cardboard pieces was 57.71% (0.70 ± 0.16% of mite infestation) at day 21 of experiment. However, the delivery of cardamon oil at the concentration of 1% and 5% (v/v) by untreated cardboard piece had 16.93% and 24.05% of efficacy to control mites. In the 2nd experiment, the application of plasma-treated cardboard pieces impregnated with 5% (v/v) clove oil induced a 38.10% reduction in the population of Varroa mites followed by 5% (v/v) of cardamon with 30% efficiency. Although, the infestation rate of Varroa in colonies was not significant different between treatments, essential oils delivered using plasma-treated cardboard pieces tended to decrease Varroa population in the treated colonies. Hence, atmospheric-pressure plasma for the modification of other materials, should be further investigated to provide alternative control treatment applications against honeybee mites