Persistence of Marine Bacterial Plasmid in the House Fly (Musca domestica): Marine-Derived Antimicrobial Resistance Genes Have a Chance of Invading the Human Environment.

The house fly is known to be a vector of antibiotic-resistant bacteria (ARB) in animal farms. It is also possible that the house fly contributes to the spread of ARB and antibiotic resistance genes (ARGs) among various environments. We hypothesized that ARB and ARGs present in marine fish and fishery food may gain access to humans via the house fly. We show herein that pAQU1, a marine bacterial ARG-bearing plasmid, persists in the house fly intestine for 5 days after fly ingestion of marine bacteria. In the case of Escherichia coli bearing the same plasmid, the persistence period exceeded 7 days. This interval is sufficient for transmission to human environments, meaning that the house fly is capable of serving as a vector of marine-derived ARGs. Time course monitoring of the house fly intestinal microflora showed that the initial microflora was occupied abundantly with Enterobacteriaceae. Experimentally ingested bacteria dominated the intestinal environment immediately following ingestion; however, after 72 h, the intestinal microflora recovered to resemble that observed at baseline, when diverse genera of Enterobacteriaceae were seen. Given that pAQU1 in marine bacteria and E. coli were detected in fly excrement (defined here as any combination of feces and regurgitated material) at 7 days post-bacterial ingestion, we hypothesize that the house fly may serve as a vector for transmission of ARGs from marine items and fish to humans via contamination with fly excrement.

Behavior and adrenal physiology of Magellanic penguins (Spheniscus magellanicus) serving as ambassador animals.

Ambassador animals are part of many zoo programs, but studies assessing their impact on these animals are relatively rare. This study validated an excrement glucocorticoid metabolite (GCM) assay for Magellanic penguins and used GCM measures in conjunction with behavioral observations to evaluate individual responses to participation in an ambassador animal program. Excrement samples and behavioral observations were collected daily from each bird during two phases, 1 week during which it participated in a twice-daily ambassador program and 1 week in which it did not. We found no differences in GCMs between phases or in comparisons between penguins with 5 or 10 years of program experience. GCM also did not show significant individual variation and did not increase over time during the program phase. There were no significant correlations between bird experience and behavior frequencies, nor GCM concentrations and behavior, across birds. We observed significant positive correlations between the penguins' engagement with novel objects during programs and their unguided approach to guests. Our results suggest that there is no adverse physiological effect of program participation on these penguins, that behavioral and physiological responses may be decoupled, and that choice and control can increase desired behaviors behavior during ambassador programs.

Problems associated with vermicomposting of dog excrement in practice using Eisenia andrei.

One 25-kg dog produces about 500 g of excrement per day. Excrement is a potentially hazardous material, as it may contain pathogenic microorganisms. Our samples were tested for the presence of thermotolerant coliform bacteria, Enterococcus spp., Escherichia coli and Salmonella spp., which are indicators of faecal contamination, as well as for the presence of helminths and their eggs. During the experiment, it was observed whether these microorganisms could be eliminated by vermicomposting. There were two variants of vermicomposting piles: one test pile (with continuous feeding) and one control pile (with a single feeding). The vermicomposting process was run in outdoor conditions in park for 51 weeks using Eisenia andrei earthworms. The vermicomposting of dog excrement with waste from park maintenance (1:2) can produce a good quality fertiliser. During the process of vermicomposting, there was a gradual decrease in the content of pathogenic bacteria. At the end of the vermicomposting process, there were no eggs or adult helminths. The vermicompost was very rich in microorganisms and enzymatic activity. The pH value was slightly alkaline, and the C:N ratio corresponded to value of mature vermicompost.

Accelerated degradation of cellulose in silkworm excrement by the interaction of housefly larvae and cellulose-degrading bacteria.

The environmental pollution caused by silkworm (Bombyx mori) excrement is prominent, and rich in refractory cellulose is the bottleneck restricting the efficient recycling of silkworm excrement. This study was performed to investigate the effects of housefly larvae vermicomposting on the biodegradation of cellulose in silkworm excrement. After six days, a 58.90% reduction of cellulose content in treatment groups was observed, which was significantly higher than 11.5% of the control groups without housefly larvae. Three cellulose-degrading bacterial strains were isolated from silkworm excrement, which were identified as Bacillus licheniformis, Bacillus amyloliquefaciens, and Bacillus subtilis based on 16S rRNA gene sequence analysis. These three bacterial stains had a high cellulose degradation index (HC value ranged to between 1.86 and 5.97 and FPase ranged from 5.07 U/mL to 7.31 U/mL). It was found that housefly larvae increased the abundance of cellulose-degrading bacterial genus (Bacillus and Pseudomonas) by regulating the external environmental conditions (temperature and pH). Carbohydrate metabolism was the bacterial communities' primary function during vermicomposting based on the PICRUSt. The results of Tax4Fun indicated that the abundance of endo-ß-1,4-glucanase and exo-ß-1,4-glucanase increased rapidly and maintained at a higher level in silkworm excrement due to the addition of housefly larvae, which contributed to the accelerated degradation of cellulose in silkworm excrement. The finding of this investigation showed that housefly larvae can significantly accelerate the degradation of cellulose in silkworm excrement by increasing the abundance of cellulose-degrading bacterial genera and cellulase.

Effects of silkworm excrement and water management on the accumulation of Cd and As in different varieties of rice and an assessment of their health risk.

Rice (Oryza sativa L.) consumption represents a major route for the exposure to cadmium (Cd) and arsenic (As) in many countries. Two varieties of rice that were grown in soils contaminated with Cd and As were evaluated for the accumulation of these toxins in rice grains and the risks of exposure of local residents to Cd and As when treated with different amounts of silkworm excrement and types of water management. Silkworm excrement, water management and the variety of rice significantly affected the accumulation of Cd and As in rice. The combination of multiple measures can be more effective at reducing heavy metals than the use of single measure, i.e., silkworm excrement management, water management, and the selection of low accumulation variety. The use of a variety that accumulates low amounts of Cd combined with 1% silkworm excrement management can effectively increase the soil pH and electrical conductivity (EC) and decrease the contents of soil available Cd and the transfer coefficients of Cd in rice, subsequently reducing the concentrations of Cd in rice grains and lowering the health risks of the intake of Cd. Similarly, the use of a conventional rice variety combined with alternating periods of drying and wetting in the three weeks before and after the heading stage decreased the contents of soil available As and the transfer coefficient of As in rice, subsequently reducing the accumulation of As in the grains and lowering the health risk of the intake of As. The significantly lower concentrations of Cd and As in rice grains and the risk of intake of Cd and As from rice was observed using a conventional rice variety combined with alternating drying-wetting in the three weeks before and after the heading stage and 1% silkworm excrement management. Thus, the combination of multiple measures in the coexistence of Cd and As in contaminated soils can be a promising strategy to avoid serious health risks and ensure the safety of food for local residents.

Shengxuening Extracted from Silkworm Excrement Mitigates Myelosuppression via SCF-Mediated JAK2/STAT3 Signaling.

Shengxuening (SXN) is a Chinese patent medicine with main ingredients (including chlorophyll derivatives and sodium iron chlorophyllin) extracted from silkworm excrement. SXN exhibited efficacy in clinical trials of renal anemia and iron deficiency anemia; however, the specific mechanisms remain unclear. This study found that SXN increased the number of peripheral blood cells and improved the bone marrow morphology in myelosuppressed mouse model, reversed the reduction in body weight and spleen indices, and increased the serum levels of erythropoietin and granulocyte-macrophage colony-stimulating factor. Quantitative real-time PCR array and Western blot analysis showed the enhanced expression of stem cell factor (SCF), JAK2, and STAT3 in the liver. These results suggested that SXN promoted the recovery of hemopoietic function in myelosuppressed models by increasing the secretion of hematopoietic factors and activating the JAK2/STAT3 pathway. Therefore, this medicine may be applied as therapeutic pharmaceutical drug to mitigate myelosuppression.

Production of Polyhydroxyalkanoates in Unsterilized Hyper-Saline Medium by Halophiles Using Waste Silkworm Excrement as Carbon Source.

The chlorophyll ethanol-extracted silkworm excrement was hardly biologically reused or fermented by most microorganisms. However, partial extremely environmental halophiles were reported to be able to utilize a variety of inexpensive carbon sources to accumulate polyhydroxyalkanoates. In this study, by using the nile red staining and gas chromatography assays, two endogenous haloarchaea strains: Haloarcula hispanica A85 and Natrinema altunense A112 of silkworm excrement were shown to accumulate poly(3-hydroxybutyrate) up to 0.23 g/L and 0.08 g/L, respectively, when using the silkworm excrement as the sole carbon source. The PHA production of two haloarchaea showed no significant decreases in the silkworm excrement medium without being sterilized compared to that of the sterilized medium. Meanwhile, the CFU experiments revealed that there were more than 60% target PHAs producing haloarchaea cells at the time of the highest PHAs production, and the addition of 0.5% glucose into the open fermentation medium can largely increase both the ratio of target haloarchaea cells (to nearly 100%) and the production of PHAs. In conclusion, our study demonstrated the feasibility of using endogenous haloarchaea to utilize waste silkworm excrement, effectively. The introduce of halophiles could provide a potential way for open fermentation to further lower the cost of the production of PHAs.

Isolation of Bacillus subtilis strain SEM-2 from silkworm excrement and characterisation of its antagonistic effect against Fusarium spp.

Fusarium wilt is a devastating soil-borne disease mainly caused by highly host-specific formae speciales of Fusarium spp. Antagonistic microorganisms play a very important role in Fusarium wilt control. Isolation of potential biocontrol strains has become increasingly important. Bacterial strain SEM-2 was isolated from the high-temperature stage of silkworm excrement composting. SEM-2 exhibited a considerable antagonistic effect against Fusarium graminearum mycelial growth and spore germination. The results of pot experiments suggested that SEM-2 has a better inhibitory effect on the early stage of disease occurrence. The green fluorescent protein labelled SEM-2 coated on the surface of tomato seeds colonised the roots of tomato plants in 15 days. Genome sequencing identified SEM-2 as a new strain of Bacillus subtilis, and genome annotation and analysis determined gene clusters related to the biosynthesis of antimicrobials, such as bacillaene, fengycin, bacillibactin, subtilosin A, surfactin, and bacilysin. Interestingly, liquid chromatography - quadrupole time-of-flight mass spectrometry revealed that metabolites in pathways associated with the synthesis of secondary metabolites and antibiotics were highly differentially expressed. These findings may help to explain the mode of action of B. subtilis SEM-2 against Fusarium spp.

Quantitative study on the fate of antibiotic emissions in China.

China, the largest producer and user of antibiotics in the world, discharges excessive amounts of these substances into the environment, without prior treatment. This results in ubiquitous distribution of these substances, as well as increased levels of drug-resistant bacteria, that will eventually cause unimaginable consequences to the environment and to humans. However, most of the research on antibiotics has focused on residue analysis of single medium such as wastewater and landfills. There is paucity of research that systematically investigates the fate of antibiotics after excretion, and specifically of end-treatment processes. In this paper, the fate of antibiotic emissions is systematically calculated. The results show that human and livestock feces account for 57.6% and 42.6% of the discharge of medicinal antibiotics and veterinary antibiotics, respectively. Of these feces types, pig feces accounted for 98.7% of antibiotic residues in livestock feces. The above conclusions can be used to clarify the direction of the tracking and supervision of antibiotic residues and provide new ideas for the treatment of antibiotics, especially their terminal removal.

[Study on degradation of silkworm excrement chemical components in vitro].

The purpose of this article is to study the degradation of chemical compositions after the silkworm excrement being expelled from the silkworm, and to determine its main metabolic compositions and their changing relationships. This research is based on UPLC-Q-TOF-MS technology. Based on the systematic analysis of the main chemical compositions contained in silkworm excrement, the principal compositions analysis(PCA) and partial least squares discriminant analysis(OPLS-DA) on commercial silkworm excrement and fresh silkworm excrement were analyzed for differences. The S-plot chart of OPLS-DA was used to select and identify the chemical compositions that contributed significantly to the difference. At the same time, the relative peak areas of the different compositions were extracted by Masslynx to obtain the relative content of different compositions in fresh silkworm excrement. The results showed that there was a significant difference in the chemical compositions between fresh silkworm excrement and commercial silkworm excrement. The difference compositions were mainly flavonoid glycosides and Diels-Alder type composition, and two types of compounds are degradated during the storage of silkworm sand. In this study, the chemical compositions of fresh silkworm excrement were systematically identified and analyzed for the first time by mass spectrometry, and it was found that some chemical compositions of silkworm excrement were degradated with time during storage.

Characterization and sequence analysis of potential biofertilizer and biocontrol agent Bacillus subtilis strain SEM-9 from silkworm excrement.

Fusarium wilt is a devastating soil-borne disease caused mainly by highly host-specific formae speciales of Fusarium oxysporum. Antagonistic microorganisms play a very important role in Fusarium wilt control, and the isolation of potential biocontrol strains is becoming more and more important. We isolated a bacterial strain (SEM-9) from the high-temperature stage of silkworm excrement composting, which had a marked ability to solubilize phosphorus, promote the growth and increase the yield of the small Chinese cabbage, and which also exhibited considerable antagonistic effect towards Fusarium sambucinum and other fungi. The result of physiological and biochemical analyses, as well as genome sequencing, showed that SEM-9 was a strain of Bacillus subtilis. Through genome annotation and analysis, it was found that SEM-9 contained genes related to the regulation of biofilm formation, which may play an important role in colonization, and gene clusters encoding the biosynthesis of antimicrobials, such as surfactin, bacilysin, fengycin, and subtilosin-A. The production of such antifungal compounds may constitute the basis of the mode-of-action of SEM-9 against Fusarium spp. These data suggested that the SEM-9 strain has potential as both a biofertilizer and a biocontrol agent, with the potential to manage Fusarium wilt disease in crops.

Recovery of active recombinant EGFP from the excrement of transgenic mice: A possible source of recombinant protein.

Transgenic animals provide attractive systems for the production of valuable recombinant proteins. Previous studies indicate that milk is a suitable source of secreted recombinant proteins. In the current study, we examine whether excrement can be another source of recombinant proteins by using transgenic mice ubiquitously-expressing green fluorescent protein (GFP) as a model. We found that the surface of excrement from GFP-transgenic mice was fluorescent, and the supernatant after centrifugation of an excrement suspension was rich in undegraded, actively fluorescing GFP. GFP was successfully purified from stool as a fluorescent 27 kDa protein by using a common procedure. Finally, we observed that the fluorescence of digested materials began in the ileum and persisted throughout the remainder of the digestive tract. Our results demonstrate that excrement, which is produced daily regardless of the sex or age of the animal, may be another feasible source of recombinant proteins. The preparation method is simple, economical, and noninvasive.

Metabolomic Analysis of Biochemical Changes in the Serum and Urine of Freund's Adjuvant-Induced Arthritis in Rats after Treatment with Silkworm Excrement.

Silkworm excrement (SE), is used as a traditional antirheumatic medicine in China. The present study was designed to investigate the therapeutic efficacy of water fraction of SE (ST) and ethanol fraction of SE (CT) at two different doses on adjuvant induced arthritis (AA) rats. Arthritis severity was evaluated by body weight, paw thickness, histological changes and index of paws oedema and spleen. Serum samples were collected for estimation of biochemical indicators and cytokines. In addition, a metabonomic method based on the ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS) had been established to investigate the holistic efficacy of SE by serum and urine. Multivariate statistical approaches, such as partial least-squares discriminant analysis (PLS-DA) and orthogonal projection to latent structures squares-discriminant analysis (OPLS-DA) were built to evaluate the therapeutic effects of SE and find potential biomarkers and metabolic pathways. Administration with SE significantly ameliorated the AA severity, including body weight loss, paw swelling, histological changes and the levels of biochemical index. 33 endogenous metabolites had been identified (10 in serum and 23 in urine) in the AA rats. Urinary and serum metabolic profiling revealed that the metabolites underpin the metabolic pathway including nicotinate and nicotinamide metabolism; pentose and glucuronate interconversions; TCA cycle; beta-Alanine metabolism; purine metabolism and glycolysis or gluconeogenesis. The altered metabolites could be regulated closer to normal level after SE intervention. The results suggested SE possesses substantial anti-arthritic activity and demonstrated that metabonomics is a powerful tool to gain insight in the mechanism of SE formula in therapy.

Catalytic liquefaction of human feces over Ni-Tm/TiO2 catalyst and the influence of operating conditions on products.

In this study, human feces were hydrothermal liquefied and converted into biocrude over Ni-Tm/TiO2 catalyst. The influence of catalysts, reaction temperature, and holding time on the distribution of products and element content of biocrude was assessed. The biocrude yield increased to 53.16% with a reaction temperature of 330 °C, a holding time of 30 min, and adding Ni-Tm/TiO2 catalyst while the liquefaction conversion peaked at 89.61%. The biocrude had an HHV of 36.64 MJ/kg and was similar to heavy crude oil. The biocrude is rich in fatty acid amides, esters, and oxygen-containing-only heteroatom-ring compounds as well as some nitrogen-containing heteroatom-ring compounds. The main gaseous products were CO2, CH4, and C2H6. Hydrothermal liquefaction over Ni-Tm/TiO2 catalyst could be a potential method to handle human excrement treatment and produce biofuel.

Coprophagy in nineteenth-century psychiatry.

This paper shows how Austrian psychiatrists of the 1870s developed the first pathological accounts of institutional coprophagia, examining how they related the behaviour to mental illness and dementia. These ideas about coprophagia contrasted dramatically to the long European pharmacological tradition of using excrement for the treatment of a wide range of health conditions. Recent medical scholarship on institutional coprophagia is also reviewed here, with a novel hypothesis proposed about why some patients in long-term care resort to the behaviour in institutions where there is little opportunity for healthy human-microbe interactions.

Hymenobacter cavernae sp. nov., isolated from a karst cave.

A novel Gram-stain-negative, rod-shaped, non-motile and red-pigmented bacterial strain, designated K1E01-27T, was isolated from an animal excrement sample which was found in a karst cave located in Guizhou province, China. The phylogenetic analysis based on 16S rRNA gene sequences indicated that strain K1E01-27T represented a member of the genus Hymenobacter within the family Cytophagaceae of the phylum Bacteroidetes. Strain K1E01-27T was most closely related to Hymenobacter algoricola VUG-A23aT, with 95.1 % 16S rRNA gene sequence similarity. Growth of strain K1E01-27T occurred at 4-35 °C, at pH 5-9 and in the presence of 0-0.5 % (w/v) NaCl. The major fatty acids were iso-C15 : 0, anteiso-C15 : 0, C16 : 1ω5c, summed feature 3 (comprising C16 : 1ω6c and/or C16 : 1ω7c) and summed feature 4 (comprising iso-C17 : 1 I/anteiso-C17 : 1 B). The major isoprenoid quinone was menaquinone 7 (MK-7). The major polar lipids were phosphatidylethanolamine, five unidentified aminophospholipids, three unidentified glycolipids, two unidentified phospholipids and one unidentified polar lipid. The DNA G+C content of the genomic DNA was 54.9 mol%. On the basis of phylogenetic, phenotypic and chemotaxonomic characteristics, strain K1E01-27T is considered to represent a novel species of the genus Hymenobacter, for which the name Hymenobacter cavernae sp. nov. is proposed. The type strain is K1E01-27T (=CGMCC 1.15197T=NBRC 112610T).

Application of DNA metabarcoding on faeces to identify European catfish Silurus glanis diet.

In this study, the results of conventional stomach-content analysis are compared with the recent DNA metabarcoding approach on faeces to identify fish species consumed by non-native European catfish Silurus glanis in the Garonne River (south-western France), with a special emphasis on anadromous prey. Fourteen prey species were identified in the stomach contents or faeces, including four anadromous fish species. Despite higher intestine than stomach emptiness, more species were identified through faecal analysis (11 of 14) than through stomach-content analysis (five of 14) suggesting that DNA metabarcoding on faeces is an efficient, non-intrusive technique to study the diet of predatory fishes.

[Silkworm excrement bacterial communities diversity in different instars based on 16S rDNA sequence analysis].

This paper investigated the diversity of the silkworm excrement bacterial communities in different ages before and after drying, aiming to clarify the differences of bacterial communities in composition and bacterial abundance and the influences of drying treatment, and provide scientific basis for the efficacy of scientific connotation and utilization of silkworm excrement. High-throughput sequencing technique was used to measure the sequence of 16S rDNA-V4 variable region of bacteria in silkworm excrement. QIIME, Mothur and PICRUSt software programs were employed to sort and calculate the number of sequences and operational taxonomic units (OTUs) for each sample. Thereafter, the abundance, distribution, alpha diversity index of species, beta diversity and bacterial communities diversity among different sample groups and predicted the bacterial gene functions were analyzed. In this study, the numbers of effective sequences for six samples were 259 250; the rarefaction curves showed a sufficient sequencing depth, and the number of OTUs was close to saturation. The bacteria in silkworm excrement belonged to the following five phylums: Proteobacteria (89.3%), Actinobacteria (5.0%), Firmicutes (4.4%), Bacteroidetes (1.1%) and Cyanobacteria (0.2%). The dominant specie was Cyanobacteria of the total bacteria identified, respectively. The abundances and diversities of the silkworm excrement bacterial communities have been reduced after drying treatment, especially the silkworm excrement of the fifth instar. PICRUSt analysis was performed to show that abundance of the functional genes such as membrane transport, carbohydrate metabolism, amino acid metabolism, lipid metabolism, nucleotide metabolism, cellular processes and signaling were relatively high. The result showed that the drying treatment could decreased the species and numbers of pathogenic bacteria in silkworm excrement obviously and improve the quality of medicinal materials. Compared with the lower ages, silkworm excrement of fifth instar seems like to be more suitable for use in medicine. Illumina MiSeq high-throughput sequencing system provides a more accurate and scientific data resource for the study of bacteria in silkworm excrement.

Evaluation of silkworm excrement and mushroom dreg for the remediation of multiple heavy metal/metalloid contaminated soil using pakchoi.

The economical, environmental friendly and efficient materials to remediate the pollution with multiple heavy metals and metalloids are scarce. Silkworm excrement (SE) and mushroom dregs (MD) are two types of agricultural wastes, and they are widely used to improve the soil fertility in many regions of China. A pot experiment with sixteen treatments was set up to assess the possibility of using SE and MD to stabilize heavy metals and metalloids and reduce their uptake in pakchoi cultivated in slightly contaminated soils with arsenic (As), cadmium (Cd), lead (Pb) and zinc (Zn). The results showed that the single addition of SE obviously stimulated the growth of pakchoi, reduced the contents of all tested heavy metals and metalloids in the edible part of pakchoi and availability of Zn and Cd in soil. The single MD treatment showed an inferior ability to enhance the growth and reduce the contents of heavy metals and metalloids in the edible part of pakchoi. The combined utilization of SE and MD appeared not to show better effects than their individual treatment when using them to remediate this contaminated soil. Some potential mechanisms on the stimulation on pakchoi growth and decreasing the accumulation of heavy metals and metalloids in pakchoi subjected to SE were suggested, including: (1) enhancing soil pH to impact the availability of heavy metals and metalloids; (2) improve the fertility of soil; (3) sulfhydryl groups of organic materials in SE play a role in conjugating heavy metals and metalloids to affect their availability in soil; and (4) stimulating the growth of pakchoi so as to show a "dilution effect" of heavy metals and metalloids.

Cultivation of earthworms and analysis of associated bacterial communities during earthworms' growth using two types of agricultural wastes.

Earthworm cultivation can effectively promote the resource utilization of agricultural waste. The efficient utilization of agricultural waste by earthworms mainly depends on the microbial communities in the guts. This study used silkworm excrement and cow manure as substrates for earthworm cultivation and investigated the associated bacterial communities during earthworms' growth. The survival rate of earthworms remained above 89% after 21 days of feeding with the two substrates. Proteobacteria, Actinobacteria, and Firmicutes constituted the predominant bacterial communities in earthworm growth, accounting for over 81% of the relative abundance in both guts and vermicompost. The bacteria richness and diversity in the foregut and midgut of earthworm were lower than those in the hindgut. The prediction function of intestinal bacterial communities of earthworms cultured with two substrates mainly involved biosynthesis, decomposition and energy production.