Acheta domesticus: A Natural Source of Anti-Skin-Aging Ingredients for Cosmetic Applications.

Acheta domesticus is an edible insect, rich in nutritional value and considered a sustainable protein source. This study aimed to investigate the potential application of A. domesticus extracts for anti-skin-aging purposes. The extracts were prepared by maceration at ambient temperature with 95% ethanol or hexane and maceration in gentle heat (45 °C) with 95% v/v ethanol or DI water. The extracts were examined for total protein, phenolic, and flavonoid contents. Protein molecular weight distribution was analyzed. The safety of the extracts was investigated in terms of irritation and cytotoxicity. Biological activities relevant to the inhibition of skin aging were evaluated, including increasing transforming growth factor-beta 1 (TGF-ß1) expression and inhibitory activities on collagenase and hyaluronidase. The aqueous extract from maceration in gentle heat had the highest total protein content (63 ± 1% w/w), total phenolic content (0.48 ± 0.03 mg GAE/g extract), TGF-ß1 stimulating activities (33 ± 2 pg/mL), and collagenase inhibition (with a half maximal inhibitory concentration of 26 ± 1 µg/mL) among various extracts investigated. It caused no irritation to the hen's egg chorioallantoic membrane and showed no cytotoxicity to human dermal fibroblasts and peripheral blood mononuclear cells. Therefore, aqueous A. domesticus extract is proposed as an innovative natural anti-skin-aging ingredient.

Optimization of Hydrolysis Conditions, Isolation, and Identification of Biologically Active Peptides Derived from Acheta domesticus for Antioxidant and Collagenase Inhibition.

The study aimed to optimize hydrolysis conditions and isolate and identify bioactive peptides with anti-skin aging effects from Acheta domesticus (house cricket). A. domesticus proteins underwent hydrolysis using Alcalase® and optimized conditions using response surface methodology through a face-centered central composite design. Variable controls (enzyme-substrate concentration (E/S), time, and temperature) were assessed for their impact on activities against collagenase, 2,2-diphenyl-1-picrylhydrazyl radical (DPPH●), and degree of hydrolysis of protein hydrolysate (PH). PH was also investigated for composition, anti-skin aging, and anti-inflammatory effects. Amino acid sequences with potent anti-skin aging activity were isolated and identified using ultrafiltration, gel filtration chromatography, and liquid chromatography coupled with tandem mass spectrometry, employing de novo sequencing. Optimal conditions for producing PH with maximum anti-skin aging activity were an E/S concentration of 2.1% (w/w), 227 min, and 61.5 °C. Glutamic acid was a predominant amino acid and PH exhibited a molecular weight below 15 kDa. Additionally, PH displayed significant activities against collagenase, hyaluronidase, DPPH●, lipid peroxidation, and NF-κB-mediated inflammation (p < 0.05). Three novel anti-skin aging peptides were identified-Ala-Val-Thr-Lys-Ala-Asp-Pro-Tyr-Thr-Asp-Gln, Thr-Val-Met-Glu-Leu-Asn-Asp-Leu-Val-Lys-Ala-Phe, and Val-Pro-Leu-Leu-Glu-Pro-Trp-exhibiting the most potent collagenase and DPPH● inhibition. Therefore, this study proposed that PH, produced with Alcalase® under optimal conditions, emerges as a promising substance with potent anti-skin aging activity for the cosmeceutical industry.

Proteomics and Immune Response Differences in Apis mellifera and Apis cerana Inoculated with Three Nosema ceranae Isolates.

Nosema ceranae infects midgut epithelial cells of the Apis species and has jumped from its original host A. cerana to A. mellifera worldwide, raising questions about the response of the new host. We compared the responses of these two species to N. ceranae isolates from A. cerana, A. mellifera from Thailand and A. mellifera from France. Proteomics and transcriptomics results were combined to better understand the impact on the immunity of the two species. This is the first combination of omics analyses to evaluate the impact of N. ceranae spores from different origins and provides new insights into the differential immune responses in honeybees inoculated with N. ceranae from original A. cerana. No difference in the antimicrobial peptides (AMPs) was observed in A. mellifera, whereas these peptides were altered in A. cerana compared to controls. Inoculation of A. mellifera or A. cerana with N. ceranae upregulated AMP genes and cellular-mediated immune genes but did not significantly alter apoptosis-related gene expression. A. cerana showed a stronger immune response than A. mellifera after inoculation with different N. ceranae isolates. N. ceranae from A. cerana had a strong negative impact on the health of A. mellifera and A. cerana compared to other Nosema isolates.

Genetic Profile of the Parasitic Varroan Mite Varroa destructor (Arachnida: Mesostigmata: Varroidae) in Taiwan: a New Taiwanese Haplotype Intermediate Between the Highly Virulent Russian and Less Virulent Japanese Types Identified in the Honey Bee Host Apis cerana.

The Modern beekeeping industry is being challenged by the varroan mite and its transmitted pathogens. Various types of Varroa destructor exhibit different levels of virulence toward honey bees, but only the Japanese (J) and Russian (R) types were found to infect Apis mellifera. Type R was more highly virulent against A. mellifera in comparison with type J. Examining the genetic profile of Varroa species is therefore of crucial importance in apiary management. In this study, maternally inherited mitochondrial cytochrome oxidase I (COI) and bisexual nuclear internal transcribed spacer (ITS) sequences of V. destructor individuals from Taiwan were determined. All 168 COI sequences observed in populations obtained from A. mellifera were identical and belonged to type J, with one base difference to that of populations collected from A. cerana; the new type is named 'T type' (Taiwan type). ITS sequences of V. destructor and its sister species V. jacobsoni were identical. A network analysis based on 611 COI sequences compiled from references indicated the presence of 27 haplotypes in V. destructor. Epidemic history and relationship analyses of V. destructor showed that the basal haplotypes were those from A. cerana and many R-extending haplotypes infesting A. mellifera involving amino acid substitutions. Calibration dating based on COI analysis revealed that V. destructor differentiated from its sibling lineage (occurring in Sri Lanka) prior to 1.3 million years ago (Mya). The ancestral haplotype retention and drift in V. destructor that occurred locally during 0.10-0.64 Mya might be relevant to its host A. cerana, which had been isolated geologically. The highly virulent type R was spreading quickly and could gradually outcompete the common and less virulent type J. Type T, being intermediate between types R and J, ought to be studied to better understand the pathogenic mechanism of V. destructor in A. mellifera. Moreover, for areas where type R does not occur, such as Taiwan, quarantine requirements are crucial for reducing invasion risks.

Tropilaelaps mercedesae Infestation Is Correlated with Injury Numbers on the Brood and the Population Size of Honey Bee Apis mellifera.

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.

Microbial community profiling and culturing reveal functional groups of bacteria associated with Thai commercial stingless worker bees (Tetragonula pagdeni).

Stingless bees play a crucial role in the environment and agriculture as they are effective pollinators. Furthermore, they can produce various products that can be exploited economically, such as propolis and honey. Despite their economic value, the knowledge of microbial community of stingless bees, and their roles on the bees' health, especially in Thailand, are in its infancy. This study aimed to investigate the composition and the functions of bacterial community associated with Tetragonula pagdeni stingless bees using culture-independent and culture-dependent approaches with emphasis on lactic acid bacteria. The culture-independent results showed that the dominant bacterial phyla were Firmicutes, Proteobacteria and Actinobacteria. The most abundant families were Lactobacillaceae and Halomonadaceae. Functional prediction indicated that the prevalent functions of bacterial communities were chemoheterotrophy and fermentation. In addition, the bacterial community might be able to biosynthesize amino acid and antimicrobial compounds. Further isolation and characterization resulted in isolates that belonged to the dominant taxa of the community and possessed potentially beneficial metabolic activity. This suggested that they are parts of the nutrient acquisition and host defense bacterial functional groups in Thai commercial stingless bees.

Preliminary Survey of Pathogens in the Asian Honey Bee (Apis cerana) in Thailand.

Widespread parasites, along with emerging threats, globalization, and climate change, have greatly affected honey bees' health, leading to colony losses worldwide. In this study, we investigated the detection of biotic stressors (i.e., viruses, microsporidian, bacteria, and fungi) in Apis cerana by surveying the colonies across different regions of Thailand (Chiang Mai in the north, Nong Khai and Khon Kaen in the northeast, and Chumphon and Surat Thani in the south, in addition to the Samui and Pha-ngan islands). In this study, we detected ABPV, BQCV, LSV, and Nosema ceranae in A. cerana samples through RT-PCR. ABPV was only detected from the samples of Chiang Mai, whereas we found BQCV only in those from Chumphon. LSV was detected only in the samples from the Samui and Pha-ngan islands, where historically no managed bees are known. Nosema ceranae was found in all of the regions except for Nong Khai and Khon Kaen in northeastern Thailand. Paenibacillus larvae and Ascosphaera apis were not detected in any of the A. cerana samples in this survey. The phylogenetic tree analysis of the pathogens provided insights into the pathogens' movements and their distribution ranges across different landscapes, indicating the flow of pathogens among the honey bees. Here, we describe the presence of emerging pathogens in the Asian honey bee as a valuable step in our understanding of these pathogens in terms of the decline in eastern honey bee populations.

Ultrasound-assisted green extraction methods: An approach for cosmeceutical compounds isolation from Macadamia integrifolia pericarp.

This study aimed was to examine the potential of several green extraction methods to extract cosmetic/cosmeceutical components from Macadamia integrifolia pericarps, which were a by-product of the macadamia nut industry. M. integrifolia pericarps were extracted by conventional solvent extraction process using 95% v/v ethanol and various green extraction methods, including infusion, ultrasound, micellar, microwave, and pulsed electric field extraction using water as a clean and green solvent. The extracts were evaluated for total phenolic content using Folin-Ciocalteu method.The antioxidant activities were evaluated by 1,1-diphenyl-2-picrylhydrazyl (DPPH), 2,2'-azinobis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), ferric reducing/antioxidant power, and ferric-thiocyanate method. The anti-skin ageing activities were investigated by means of collagenase, elastase, and hyaluronidase inhibition using enzyme-substrate reaction assay. The irritation profile of the extracts was evaluated by the hen's egg test-chorioallantoic membrane (HET-CAM) test. The results noted that ultrasound-assisted extraction yielded the significantly highest extract amount with the significantly highest total phenolic content (p < 0.05), especially when the extraction time was 10 min. The aqueous extract from ultrasound-assisted extraction possessed the most potent antioxidant and anti-skin ageing activities (p < 0.05). Its antioxidant activities were comparable to ascorbic acid and Trolox, whereas the anti-skin ageing activities were equivalent to epigallocatechin-3-gallate and oleanolic acid. Besides, the extract was safe since it induced no irritation in the HET-CAM test. Therefore, ultrasound-assisted extraction was suggested as an environmentally friendly extraction method for M. integrifolia pericarp extraction and further application in the cosmetic/cosmeceutical industries.

Living Fungi in an Opencast Limestone Mine: Who Are They and What Can They Do?

Opencast limestone mines or limestone quarries are considered challenging ecosystems for soil fungi as they are highly degraded land with specific conditions, including high temperature, prolonged sunlight exposure, and a lack of organic matter, moisture, and nutrients in soil. In such ecosystems, certain fungi can survive and have a crucial function in maintaining soil ecosystem functions. Unfortunately, we know very little about taxonomic diversity, potential functions, and the ecology of such fungi, especially for a limestone quarry in a tropical region. Here, we characterized and compared the living soil fungal communities in an opencast limestone mine, including mining site and its associated rehabilitation site (9 months post-rehabilitation), with the soil fungal community in a reference forest, using the amplicon sequencing of enrichment culture. Our results showed that living fungal richness in the quarry areas was significantly lower than that in the reference forest, and their community compositions were also significantly different. Living fungi in the mining sites mostly comprised of Ascomycota (Eurotiomycetes and Sordariomycetes) with strongly declined abundance or absence of Basidiomycota and Mucoromycota. After nine months of rehabilitation, certain taxa were introduced, such as Hypoxylon spp. and Phellinus noxius, though this change did not significantly differentiate fungal community composition between the mining and rehabilitation plots. The majority of fungi in these plots are classified as saprotrophs, which potentially produce all fifteen soil enzymes used as soil health indicators. Network analysis, which was analyzed to show insight into complex structures of living fungal community in the limestone quarry, showed a clear modular structure that was significantly impacted by different soil properties. Furthermore, this study suggests potential taxa that could be useful for future rehabilitation.

Interaction between Thiamethoxam and Deformed Wing Virus Type A on Wing Characteristics and Expression of Immune and Apoptosis Genes in Apis mellifera.

Honey bees are economically important insects for crop pollination. They play a significant role as pollinators of wild plants and agricultural crops and produce economical products, such as honey, royal jelly, wax, pollen, propolis, and venom. Despite their ecological and economical importance, the global honey bee population is in decline due to factors including pathogens, parasites, intensive agriculture, and pesticides. Moreover, these factors may be interlinked and exacerbate the loss of honey bees. This study aimed to investigate the interaction between a pesticide, thiamethoxam, and deformed wing virus type A (DWV-A) to honey bees and the effects on survival rate, wing characteristics, and expression of immune and apoptosis genes in Apis mellifera. We described the potential interaction between thiamethoxam and DWV-A on honey bee wing characteristics, DWV-A loads, and the expressions of immune (defensin, abaecin, and hymenoptaecin) and apoptosis genes (buffy, apaf1, caspase3-like, caspase8-like, and caspase9-like). Honey bee larvae were fed with three different thiamethoxam doses (0.001, 1.4, and 14.3 ng/µL of the diet). Then, thiamethoxam-treated white-eyed pupae were injected with 107 copy numbers/honey bee of the DWV-A genome. The interaction between thiamethoxam and DWV-A caused a high mortality rate, crippled wings in newly emerged adult honey bees (100%), and resulted in induced expression of hymenoptaecin gene compared to the control group, while downregulation of caspase8-like, caspase9-like genes compared to the DWV injection group. Therefore, the potential interaction between thiamethoxam and DWV-A might have a deleterious effect on honey bee lifespan. The results from this study could be used as a tool to combat DWV-A infection and mitigate pesticide usage to alleviate the decrease in the honey bee population.

Identification of Microorganisms Dwelling on the 19th Century Lanna Mural Paintings from Northern Thailand Using Culture-Dependent and -Independent Approaches.

Lanna painting is a unique type of painting in many temples in the Northern Thai region. Similar to most mural paintings, they usually decay over time partly due to the activity of microbes. This study aimed to investigate the microorganisms from two Lanna masterpiece paintings in two temples that differ in the numbers of visitors using both culture-dependent and -independent approaches. The microorganisms isolated from the murals were also tested for the biodeterioration activities including discoloration, acid production and calcium precipitation. Most microorganisms extracted from the paintings were able to discolor the paints, but only fungi were able to discolor, produce acids and precipitate calcium. The microorganism communities, diversity and functional prediction were also investigated using the culture-independent method. The diversity of microorganisms and functional prediction were different between the two temples. Gammaproteobacteria was the predominant group of bacteria in both temples. However, the fungal communities were different between the two temples as Aspergillus was the most abundant genus in the site with higher number of visitors [Buak Krok Luang temple (BK)]. Conversely, mural paintings at Tha Kham temple (TK) were dominated by the Neodevriesia genera. We noticed that a high number of visitors (Buak Krok Luang) was correlated with microbial contamination from humans while the microbial community at Tha Kham temple had a higher proportion of saprotrophs. These results could be applied to formulate a strategy to mitigate the amount of tourists as well as manage microorganism to slow down the biodeterioration process.

Bacterial Communities in Lanna Phak-Gard-Dong (Pickled Mustard Green) from Three Different Ethnolinguistic Groups in Northern Thailand.

The Lanna region, the main part of northern Thailand, is a place of ethnic diversity. In this study, we investigated phak-gard-dong (PGD), or pickled mustard green (Brassica juncea L. Czern.), for its beneficial bacteria content and to analyse the variations in bacterial compositions among the PGD of three different ethnolinguistic groups, the Karen, Lawa, and Shan. DNA was extracted from the PGD pickled brine, and 16S rRNA gene Illumina sequencing was performed. Metagenomic data were analysed and the results demonstrated that the dominant bacterial species were Weissella (54.2%, 65.0%, and 10.0%) and Lactobacillus (17.5%, 5.6%, and 79.1%) in the PGD of the Karen, Lawa, and Shan, respectively. Pediococcus was found only in the PGD of the Karen and Shan. Bacterial communities in PGD of the Lawa were distinctive from the other ethnic groups, both in the alpha and beta diversity, as well as the predicted functions of the bacterial communities. In addition, overall network analysis results were correlated to bacterial proportions in every ethnic PGD. We suggest that all ethnic PGDs have the potential to be a good source of beneficial bacteria, warranting its conservation and further development into health food products.

Anti-Inflammatory and Antimicrobial Activities of Fermented Ocimum sanctum Linn. Extracts against Skin and Scalp Microorganisms.

This study aimed to revealed anti-inflammatory and antimicrobial activities of fermented Ocimum sanctum Linn. (FE). The fermentation process with Lactobacillus plantarum was compared with the solvent extraction methods. Antimicrobial activity against the growth of Staphylococcus aureus, Staphylococcus epidermidis, Propionibacterium acnes, Candida albicans, and Malassezia furfur was investigated via broth dilution method. High performance thin layer chromatography was used to determine eugenol content. The anti-inflammation was investigated by means of nuclear factor kappa B (NF-κB) expression inhibition by Western blot analysis. FE yielded the highest amount (11.93 % w/w), the highest eugenol content (39.3±12.6 % w/w), and the highest antimicrobial activities comparing to the extracts obtained from the solvent extractions. The fungal inhibition against M. furfur 656 was equivalent to that of ketoconazole. Furthermore, the bacterial inhibition on S. aureus and S. epidermidis was compared to that of Penicillin G at minimum inhibitory concentration (MIC) of 0.125 mg/mL and 0.25 mg/mL, respectively. Interestingly, FE had lower MIC and minimum bactericidal concentration against P. acnes than Penicillin G and also possessed comparable anti-inflammatory activity to indomethacin with the NF-κB suppression of 42.7±4.6 %. Therefore, FE are potentially natural anti-inflammation and antimicrobial agents for topical applications in the pharmaceutical and cosmetic industries.

Comparative Study of Antimicrobial Properties of Bee Venom Extracts and Melittins of Honey Bees.

Bee venom (BV), or apitoxin, is a complex substance produced by a gland in the abdominal cavity of bees. The main component of BV is melittin, which is a largely studied substance due to its biological properties. To date, the most well-known bee venom and melittin are derived from domesticated honey bees, while venom and melittin derived from wild honey bees have been under-investigated. Hence, this study primarily reports the antimicrobial activities of bee venom and synthetic melittin derived from four different honey bee species (Apis mellifera, A. cerana, A. dorsata, and A. florea) in Thailand. All the bee venom extracts and melittins showed more robust antibacterial activities against Gram-positive (Bacillus subtilis, Micrococcus luteus, Staphylococcus aureus, S. aureus MRSA, and S. epidermidis) than Gram-negative bacteria (Escherichia coli, Klebsiella pneuminiae, and Salmonella typhimurium) or a fungus (Candida albicans), while the synthetic melittins also have antimicrobial activity at higher concentrations than the bee venom extract. Furthermore, the A. cerana venom extract showed the highest activity against the tested bacteria, followed by A. mellifera, A. florea, and A. dorsata. Therefore, A. cerana venom may be further developed for use in medical applications as a potential alternative agent against Gram-positive bacteria and antibiotic-resistant bacteria.

Potentiality of Melittin-Loaded Niosomal Vesicles Against Vancomycin-Intermediate Staphylococcus aureus and Staphylococcal Skin Infection.

BACKGROUND: Staphylococcus aureus is an important human pathogen, especially causing skin and soft tissue infections (SSTIs). Over the decades, the infections caused by antibiotic-resistant strains have often become life-threatening. Consequently, exploration and development of competent approaches to combat these serious circumstances are urgently required. METHODS: The antibacterial activity of melittin (Mel) on S. aureus, methicillin-resistant S. aureus (MRSA) and clinical isolates of vancomycin-intermediate S. aureus (VISA) was investigated by minimum inhibitory concentration (MIC) and time-killing assays. The localization of Mel on the bacterial cell was visualized by confocal laser scanning microscopy and its effect on the membrane was indicated based on propidium iodide uptake. The non-ionic surfactant vesicle (NISV) or niosome nanocarrier was established for Mel loading (Mel-loaded NISV) by the thin-film hydration method. Physicochemical and in vitro biological properties of Mel-loaded NISVs were characterized. The cellular uptake of Mel-loaded NISVs was evaluated by holotomography analysis. In addition, an ex vivo study was conducted on a porcine ear skin model to assess the permeation ability of Mel-loaded NISVs and their potential to inhibit bacterial skin infection. RESULTS: The effective inhibitory activity of Mel on skin pathogens was demonstrated. Among the tested strains, VISA was most susceptible to Mel. Regarding to its function, Mel targeted the bacterial cell envelope and disrupted cell membrane integrity. Mel-loaded NISVs were successfully fabricated with a nano-size of 120-200 nm and entrapment efficiency of greater than 90%. Moreover, Mel-loaded NISVs were taken up and accumulated in the intracellular space. Meanwhile, Mel was released and distributed throughout the cytosol and nucleus. Mel-loaded NISVs efficiently inhibited the growth of bacteria, particularly MRSA and VISA. Importantly, they not only penetrated epidermal and dermal skin layers, but also reduced the bacterial growth in infected skin. CONCLUSION: Mel-loaded NISVs have a great potential to exhibit antibacterial activity. Therapeutic application of Mel-loaded NISVs could be further developed as an alternative platform for the treatment of skin infection via dermal and transdermal delivery.

Skin irritation and potential antioxidant, anti-collagenase, and anti-elastase activities of edible insect extracts.

This study aimed to investigate antioxidant, anti-aging, and irritation properties of Thai edible insect extracts, including Bombyx mori, Omphisa fuscidentalis, Euconocephalus sp., Patanga succincta, Acheta domesticus, and Lethocerus indicus. Insects were extracted by 2 different methods, including maceration using ethanol or hexane and digestion using DI water. Then the extracts were determined for protein content using bicinchoninic acid assay and antioxidant activities using 2,2'-azinobis (3-ethylbenzothiazoline-6-sulfonic acid), 2,2-diphenyl-1-picrylhydrazyl, ferric reducing antioxidant power, and ferric thiocyanate assays. Anti-aging activities were investigated by determination of collagenase and elastase inhibitory activities using spectrophotometric assay. Maceration by hexane yielded the highest extract content, whereas aqueous extract from digestion possessed the significantly highest protein content and biological activities (p < 0.05). Interestingly, aqueous extracts of A. domesticus possessed the significantly highest biological activities (p < 0.05) with Trolox equivalent antioxidant capacity value of 8.8 ± 0.1 mmol Trolox/mg, DPPH· inhibition of 19.5 ± 3.8%, equivalent concentration of 12.1 ± 0.7 µM FeSO4/mg, lipid peroxidation inhibition of 31.3 ± 2.4%, collagenase inhibition of 60.8 ± 2.1%, elastase inhibition of 17.0 ± 0.1%, and no irritation effect on chorioallantoic membrane and volunteers. Therefore, aqueous extract of A. domesticus would be suggested for further topical product development.

Impact of Nosema Disease and American Foulbrood on Gut Bacterial Communities of Honeybees Apis mellifera.

Honeybees, Apis mellifera, are important pollinators of many economically important crops. However, one of the reasons for their decline is pathogenic infection. Nosema disease and American foulbrood (AFB) disease are the most common bee pathogens that propagate in the gut of honeybees. This study investigated the impact of gut-propagating pathogens, including Nosema ceranae and Paenibacillus larvae, on bacterial communities in the gut of A. mellifera using 454-pyrosequencing. Pyrosequencing results showed that N. ceranae was implicated in the elimination of Serratia and the dramatic increase in Snodgrassella and Bartonella in adult bees' guts, while bacterial communities of P. larvae-infected larvae were not affected by the infection. The results indicated that only N. ceranae had an impact on some core bacteria in the gut of A. mellifera through increasing core gut bacteria, therefore leading to the induction of dysbiosis in the bees' gut.

The genomic basis of evolutionary differentiation among honey bees.

In contrast to the western honey bee, Apis mellifera, other honey bee species have been largely neglected despite their importance and diversity. The genetic basis of the evolutionary diversification of honey bees remains largely unknown. Here, we provide a genome-wide comparison of three honey bee species, each representing one of the three subgenera of honey bees, namely the dwarf (Apis florea), giant (A. dorsata), and cavity-nesting (A. mellifera) honey bees with bumblebees as an outgroup. Our analyses resolve the phylogeny of honey bees with the dwarf honey bees diverging first. We find that evolution of increased eusocial complexity in Apis proceeds via increases in the complexity of gene regulation, which is in agreement with previous studies. However, this process seems to be related to pathways other than transcriptional control. Positive selection patterns across Apis reveal a trade-off between maintaining genome stability and generating genetic diversity, with a rapidly evolving piRNA pathway leading to genomes depleted of transposable elements, and a rapidly evolving DNA repair pathway associated with high recombination rates in all Apis species. Diversification within Apis is accompanied by positive selection in several genes whose putative functions present candidate mechanisms for lineage-specific adaptations, such as migration, immunity, and nesting behavior.

Differential Viral Distribution Patterns in Reproductive Tissues of Apis mellifera and Apis cerana Drones.

Honeybee drones are male bees that mate with virgin queens during the mating flight, consequently transferring their genes to offspring. Therefore, the health of drones affects the overall fitness of the offspring and ultimately the survivability of the colony. Honeybee viruses are considered to be a major threat to the health of honeybees. In the present study, we demonstrated the pattern of common honeybee viruses in various tissues of drones in the western honeybee, Apis mellifera, and the eastern honeybee, Apis cerana. Drones were collected during the mating flight and analyzed using quantitative real-time (qRT-PCR) to detect the presence of seven honeybee viruses. The qRT-PCR result revealed that three honeybee viruses, namely Black Queen Cell Virus (BQCV), Deformed Wing Virus (DWV), and Chinese Sacbrood Virus (CSBV), were detected in the reproductive tissues of A. mellifera and A. cerana drones. The results from qRT-PCR showed that the Israeli Acute Paralysis Virus (IAPV) was only detected in A. mellifera drone body tissues. Moreover, the prevalence of DWV and BQCV in the drones collected from A. mellifera colonies was significantly higher than that of A. cerana. In addition, virus multiple infections were higher in A. mellifera drones compared to those in A. cerana. CSBV was found predominantly in the reproductive tissues of A. cerana drones. This study is the first report describing the presence of the CSBV in reproductive tissues of A. mellifera drones. Our results may reflect the preference of honeybee viruses in honeybee species and may provide a piece of interesting evidence for understanding the virus transmission in A. cerana.

Soil bacterial communities and their associated functions for forest restoration on a limestone mine in northern Thailand.

Opencast mining removes topsoil and associated bacterial communities that play crucial roles in soil ecosystem functioning. Understanding the community composition and functioning of these organisms may lead to improve mine-rehabilitation practices. We used a culture-dependent method, combined with Illumina sequencing, to compare the taxonomic richness and composition of living bacterial communities in opencast mine substrates and young mine-rehabilitation plots, with those of soil in adjacent remnant forest at a limestone mine in northern Thailand. We further investigated the effects of soil physico-chemical factors and ground-flora cover on the same. Although, loosened subsoil, brought in to initiate rehabilitation, improved water retention and facilitated plant re-establishment, it did not increase the population density of living microbes substantially within 9 months. Planted trees and sparse ground flora in young rehabilitation plots had not ameliorated the micro-habitat enough to change the taxonomic composition of the soil bacteria compared with non-rehabilitated mine sites. Viable microbes were significantly more abundant in forest soil than in mine substrates. The living bacterial community composition differed significantly, between the forest plots and both the mine and rehabilitation plots. Proteobacteria dominated in forest soil, whereas Firmicutes dominated in samples from both mine and rehabilitation plots. Although, several bacterial taxa could survive in the mine substrate, soil ecosystem functions were greatly reduced. Bacteria, capable of chitinolysis, aromatic compound degradation, ammonification and nitrate reduction were all absent or rare in the mine substrate. Functional redundancy of the bacterial communities in both mine substrate and young mine-rehabilitation soil was substantially reduced, compared with that of forest soil. Promoting the recovery of microbial biomass and functional diversity, early during mine rehabilitation, is recommended, to accelerate soil ecosystem restoration and support vegetation recovery. Moreover, if inoculation is included in mine rehabilitation programs, the genera: Bacillus, Streptomyces and Arthrobacter are likely to be of particular interest, since these genera can be cultivated easily and this study showed that they can survive under the extreme conditions that prevail on opencast mines.