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First Report of Damping-Off on Basella rubra Caused by Rhizoctonia solani Anastomosis Group 4 in Florida

Liao X, Fu Y, Zhang S, Duan YP

Plant Dis, 2012 Feb;96(2):288.
PMID: 30731824 DOI: 10.1094/PDIS-08-11-0639

Indian spinach (Basella rubra L.) is a red stem species of Basella that is cultivated worldwide as an ornamental and the aerial parts are also consumed as a vegetable. In May of 2011, symptoms of damping-off were observed on approximately 10% of the plants at the stem base around the soil line of seedlings in a greenhouse in Homestead, FL. Lesions were initially water soaked, grayish to dark brown, irregular in shape, and sunken in appearance on large plants, causing the infected seedlings to collapse and eventually die. Symptomatic stem tissue was surface sterilized with 0.6% sodium hypochlorite, rinsed in sterile distilled water, air dried, and plated on potato dextrose agar (PDA). Plates were incubated at 25°C in darkness for 3 to 5 days. A fungus was isolated in all six isolations from symptomatic tissues on PDA. Fungal colonies on PDA were light gray to brown with abundant growth of mycelia, and the hyphae tended to branch at right angles when examined under a microscope. A septum was always present in the branch of hyphae near the originating point and a slight constriction at the branch was observed. Neither conidia nor conidiophores were found from the cultures on PDA. The characteristics of hyphae, especially the right angle branching of mycelia, indicate close similarity to those of Rhizoctonia solani (2,3). The internal transcribed spacer (ITS) region of rDNA was amplified with the primers ITS1/ITS4 and sequenced (GenBank Accession No. JN545836). Subsequent database searches by the BLASTN program indicated that the resulting sequence had a 100% identity over 472 bp with the corresponding gene sequence of R. solani anastomosis group (AG) 4 (GenBank Accession No. JF701752.1), a fungal pathogen reported to cause damping-off on many crops. Pathogenicity was confirmed through inoculation of healthy India spinach plants with the hyphae of isolates. Four 4-week-old plants were inoculated with the isolates by placing a 5-mm PDA plug of mycelia at the stem base and covering with a thin layer of the soil. Another four plants treated with sterile PDA served as a control. After inoculation, the plants were covered with plastic bags for 24 h and maintained in a greenhouse with ambient conditions. Four days after inoculation, water-soaked, brown lesions, identical to the symptoms described above, were observed on the stem base of all inoculated plants, whereas no symptoms developed on the control plants. The fungus was isolated from affected stem samples, and the identity was confirmed by microscopic appearance of the hyphae and sequencing the ITS1/ITS4 intergenic spacer region, fulfilling Koch's postulates. This pathogenicity test was conducted twice. R. solani has been reported to cause damping-off of B. rubra in Ghana (1) and Malaysia (4). To our knowledge, this is the first report of damping-off caused by R. solani AG-4 on Indian spinach in Florida and the United States. With the increased interest in producing Asian vegetables for food and ornamental purposes, the occurrence of damping-off on Indian spinach needs to be taken into account when designing programs for disease management in Florida. References: (1) H. A. Dade. XXIX. Bull. Misc. Inform. 6:205, 1940. (2) J. R. Parmeter et al. Phytopathology 57:218, 1967. (3) B. Sneh et al. Identification of Rhizoctonia species. The American Phytopathological Society, St Paul, MN, 1991. (4) T. H. Williams and P. S. W. Liu. Phytopathol. Pap. 19:1, 1976.
In vitro assessment on effect of duodenal contents on the lead (Pb(2+)) binding capacity of two probiotic bacterial strains

Xing S, Song Y, Liang JB, Faseleh Jahromi M, Shokryazda P, Mi J, et al.

Ecotoxicol Environ Saf, 2017 May;139:78-82.
PMID: 28113114 DOI: 10.1016/j.ecoenv.2017.01.016

In vitro Lead (Pb(2+)) binding capacity of two probiotic bacteria strains, namely Bifidobacterium longumBB79 and Lactobacillus pentosusITA23, was assessed following incubation with the intestinal contents (IC) of laying hens. Results of this study demonstrated that IC treatment significantly enhanced (P<0.01) Pb(2+) binding capacity of both bacterial strains. Fourier transform infrared analysis indicated that several functional groups (O-H or N-H, C-H, C˭O, C-O, and C-O-C) on the bacteria cell wall involved in metal ion binding were altered after IC incubation, and new groups appeared between the 3700cm(-1) and 4000cm(-1)bands. Transmission electron microscopy demonstrated that after incubation with IC, unidentified IC components created new binding sites on the bacterial cell surface. These particles also changed the mechanism of Pb(2+) binding of the two strains from intracellular accumulation to extracellular adsorption.
Fulltext Exploratory Analysis of the Microbiological Potential for Efficient Utilization of Fiber Between Lantang and Duroc Pigs

Cheng P, Wang Y, Liang J, Wu Y, Wright A, Liao X

Front Microbiol, 2018;9:1342.
PMID: 29988353 DOI: 10.3389/fmicb.2018.01342

There is growing interest in the use of unconventional feed ingredients containing higher dietary fiber for pig production due to increasing prices of cereal grains and the potential health benefits of dietary fiber on host animals. This study aimed to gain insight into the community-wide microbiome population between the Chinese native Lantang pigs and the commercial Duroc pigs to uncover the microbiological mechanisms for the degradation capacity of fiber in pigs. Utilizing the metagenomics approach, we compared the phylogeny and functional capacity of the fecal microbiome from approximately 150-day-old female Lantang and Duroc pigs fed a similar diet. The structure of the fecal microbial community from the two pig breeds was different at the genus level; the number of genes associated with fiber degradation was higher in Lantang pigs. Further analysis and prediction of their functions from the fecal microbiomes of the two pig breeds revealed that the degradation capacities of fiber, branched chain fatty acids, and oligosaccharides were higher in Lantang pigs. The ability of lignocellulose bonding modules and the transport capacities of xylose, L-arabinose, ribose and methyl galactose were also higher in Lantang pigs. Similarly, the metabolic capacities of xylose, ribose, and fucose and the potential effectiveness of the tricarboxylic acid cycle (TCA) and gene abundance in the hydrogen sink pathway were higher in the fecal microbiome from Lantang pigs. Lantang pigs have a higher capacity to utilize dietary fiber than Duroc pigs, and the differences in the capability to utilize dietary fiber between the indigenous and commercial pigs could be differences in the composition and biological function of the gut microbiota.
Lead biosorption of probiotic bacteria: effects of the intestinal content from laying hens

Xing S, Wang J, Liang JB, Jahromi MF, Zhu C, Shokryazdan P, et al.

Environ Sci Pollut Res Int, 2017 May;24(15):13528-13535.
PMID: 28390025 DOI: 10.1007/s11356-017-8896-6

This study investigated the effects and the possible mechanisms of intestinal content (IC) from laying hens on in vitro lead (Pb(2+)) biosorption of four probiotic bacterial strains (Bifidobacterium longum BB79, Lactobacillus paracasei Kgl6, Lactobacillus pentosus ITA23, and Lactobacillus acidipiscis ITA44). The total Pb(2+) removal capacity of the four probiotic strains, with and without capsule polysaccharides (CPSs), increased in the presence of IC compared to the control (without IC). SEM imaging revealed certain unidentified particles from the IC adhered on the surface of bacterial cells sorted out using flow cytometry. Follow-up experiment showed an overall trend of increase in the Pb(2+) removal capacity of the sorted bacteria, but statistically significant for L. pentosus ITA23 and B. longum BB79 after incubation with IC, particularly with the suspended solid portion of the IC. In addition, the Fourier transform infrared spectrophotometer data showed that functional groups such as C-H, O-H, C=O, and C-O-C which possibly associated with Pb(2+) binding were mainly presented in the suspended solid portion of IC. Putting the above together, we postulated that the enhanced Pb(2+) binding capacity the probiotic bacteria incubated in IC is due to the adherence of the yet to be identified particles which could much exist in suspended solid portion of IC containing negatively charged functional groups which bind with the positive Pb(2+) ions.
Mode of action of Saccharomyces cerevisiae in enteric methane mitigation in pigs

Gong YL, Liang JB, Jahromi MF, Wu YB, Wright AG, Liao XD

Animal, 2018 Feb;12(2):239-245.
PMID: 28735588 DOI: 10.1017/S1751731117001732

The objectives of this study were to determine the effect and mode of action of Saccharomyces cerevisiae (YST2) on enteric methane (CH4) mitigation in pigs. A total of 12 Duroc×Landrace×Yorkshire male finisher pigs (60±1 kg), housed individually in open-circuit respiration chambers, were randomly assigned to two dietary groups: a basal diet (control); and a basal diet supplemented with 3 g/YST2 (1.8×1010 live cells/g) per kg diet. At the end of 32-day experiment, pigs were sacrificed and redox potential (Eh), pH, volatile fatty acid concentration, densities of methanogens and acetogens, and expression of methyl coenzyme-M reductase subunit A gene were determined in digesta contents from the cecum, colon and rectum. Results showed that S. cerevisiae YST2 decreased (P<0.05) the average daily enteric CH4 production by 25.3%, lowered the pH value from 6.99 to 6.69 in the rectum, and increased the Eh value in cecum and colon by up to -55 mV (P<0.05). Fermentation patterns were also altered by supplementation of YST2 as reflected by the lower acetate, and higher propionate molar proportion in the cecum and colon (P<0.05), resulting in lower acetate : propionate ratio (P<0.05). Moreover, there was a 61% decrease in Methanobrevibacter species in the upper colon (P<0.05) and a 19% increase in the acetogen community in the cecum (P<0.05) of treated pigs. Results of our study concluded that supplementation of S. cerevisiae YST2 at 3 g/kg substantially decreased enteric CH4 production in pigs.
Effects of fermented soybean meal on carbon and nitrogen metabolisms in large intestine of piglets

Zhang YT, Lu DD, Chen JY, Yu B, Liang JB, Mi JD, et al.

Animal, 2018 Oct;12(10):2056-2064.
PMID: 29362016 DOI: 10.1017/S1751731118000058

Fermented soybean meal (FSM), which has lower anti-nutritional factors and higher active enzyme, probiotic and oligosaccharide contents than its unfermented form, has been reported to improve the feeding value of soybean meal, and hence, the growth performance of piglets. However, whether FSM can affect the bacterial and metabolites in the large intestine of piglets remains unknown. This study supplemented wet-FSM (WFSM) or dry-FSM (DFSM) (5% dry matter basis) in the diet of piglets and investigated its effects on carbon and nitrogen metabolism in the piglets' large intestines. A total of 75 41-day-old Duroc×Landrace×Yorkshire piglets with an initial BW of 13.14±0.22 kg were used in a 4-week feeding trial. Our results showed that the average daily gain of piglets in the WFSM and DFSM groups increased by 27.08% and 14.58% and that the feed conversion ratio improved by 18.18% and 7.27%, respectively, compared with the control group. Data from the prediction gene function of Phylogenetic Investigation of Communities by Reconstruction of Unobserved States (PICRUSt) based on 16S ribosomal RNA (rRNA) sequencing showed that carbohydrate metabolism function families in the WFSM and DFSM groups increased by 3.46% and 2.68% and that the amino acid metabolism function families decreased by 1.74% and 0.82%, respectively, compared with the control group. These results were consistent with those of other metabolism studies, which showed that dietary supplementation with WFSM and DFSM increased the level of carbohydrate-related metabolites (e.g. 4-aminobutanoate, 5-aminopentanoate, lactic acid, mannitol, threitol and β-alanine) and decreased the levels of those related to protein catabolism (e.g. 1,3-diaminopropane, creatine, glycine and inosine). In conclusion, supplementation with the two forms of FSM improved growth performance, increased metabolites of carbohydrate and reduced metabolites of protein in the large intestine of piglets, and WFSM exhibited a stronger effect than DFSM.
Fulltext The draft genomes of five agriculturally important African orphan crops

Chang Y, Liu H, Liu M, Liao X, Sahu SK, Fu Y, et al.

Gigascience, 2019 03 01;8(3).
PMID: 30535374 DOI: 10.1093/gigascience/giy152

BACKGROUND: The expanding world population is expected to double the worldwide demand for food by 2050. Eighty-eight percent of countries currently face a serious burden of malnutrition, especially in Africa and south and southeast Asia. About 95% of the food energy needs of humans are fulfilled by just 30 species, of which wheat, maize, and rice provide the majority of calories. Therefore, to diversify and stabilize the global food supply, enhance agricultural productivity, and tackle malnutrition, greater use of neglected or underutilized local plants (so-called orphan crops, but also including a few plants of special significance to agriculture, agroforestry, and nutrition) could be a partial solution.
RESULTS: Here, we present draft genome information for five agriculturally, biologically, medicinally, and economically important underutilized plants native to Africa: Vigna subterranea, Lablab purpureus, Faidherbia albida, Sclerocarya birrea, and Moringa oleifera. Assembled genomes range in size from 217 to 654 Mb. In V. subterranea, L. purpureus, F. albida, S. birrea, and M. oleifera, we have predicted 31,707, 20,946, 28,979, 18,937, and 18,451 protein-coding genes, respectively. By further analyzing the expansion and contraction of selected gene families, we have characterized root nodule symbiosis genes, transcription factors, and starch biosynthesis-related genes in these genomes.
CONCLUSIONS: These genome data will be useful to identify and characterize agronomically important genes and understand their modes of action, enabling genomics-based, evolutionary studies, and breeding strategies to design faster, more focused, and predictable crop improvement programs.
The health status and health needs of Chinese population in Glasgow

Liao XH, McIlwaine G

Scott Med J, 1995 Jun;40(3):77-80.
PMID: 7569869

This cross sectional study explored the health problems and health needs in the local Chinese community in Glasgow. Several data collection methods have been used in this study, including face-to-face and telephone structured interviews, postal and hand delivered questionnaires. A total of 800 questionnaires were processed, and 493 were completed, giving an overall response rate 61.6%. The results from the present survey indicated that the health status of Chinese residents in Glasgow is poorer than that of the local population. The most important findings of the study is that the Chinese community in Glasgow underuse health services, and unmet health needs exist in the community. The main barrier to effective use of present health services and benefit from the health promotion and health education programmes is language difficulties. Following discussion with the local community, options for improving the health services for the Chinese community in Glasgow were obtained. The findings of the study have implications for health service purchaser/providers of health care to the Chinese population generally in Scotland.
Fulltext Potentially toxic elements in human scalp hair around China's largest polymetallic rare earth ore mining and smelting area

Dai L, Deng L, Wang W, Li Y, Wang L, Liang T, et al.

Environ Int, 2023 Feb;172:107775.
PMID: 36739854 DOI: 10.1016/j.envint.2023.107775

There is a growing concern about human health of residents living in areas where mining and smelting occur. In order to understand the exposure to the potentially toxic elements (PTEs), we here identify and examine the cadmium (Cd), chromium (Cr), copper (Cu), manganese (Mn), nickel (Ni), lead (Pb) and zinc (Zn) in scalp hair of residents living in the mining area (Bayan Obo, n = 76), smelting area (Baotou, n = 57) and a reference area (Hohhot, n = 61). In total, 194 hair samples were collected from the volunteers (men = 87, women = 107) aged 5-77 years old in the three areas. Comparing median PTEs levels between the young and adults, Ni levels were significantly higher in adults living in the smelting area while Cr was highest in adults from the mining area, no significant difference was found for any of the elements in the reference area. From the linear regression model, no significant relationship between PTEs concentration, log10(PTEs), and age was found. The concentrations of Ni, Cd, and Pb in hair were significantly lower in the reference area when compared to both mining and smelting areas. In addition, Cu was significantly higher in the mining area when compared to the smelting area. Factor analysis (FA) indicated that men and women from the smelting area (Baotou) and mining area (Bayan Obo), respectively, had different underlying communality of log10(PTEs), suggesting different sources of these PTEs. Multiple factor analysis quantilized the importance of gender and location when combined with PTEs levels in human hair. The results of this study indicate that people living in mining and/or smelting areas have significantly higher PTEs (Cu, Ni, Cd, and Pb) hair levels compared to reference areas, which may cause adverse health effects. Remediation should therefore be implemented to improve the health of local residents in the mining and smelting areas.
Sodium butyrate mitigates in vitro ammonia generation in cecal content of laying hens

Wang A, Wang Y, Di Liao X, Wu Y, Liang JB, Laudadio V, et al.

Environ Sci Pollut Res Int, 2016 Aug;23(16):16272-9.
PMID: 27154844 DOI: 10.1007/s11356-016-6777-z

One of the environmental challenges that modern poultry industry faced is odor pollution caused by ammonia emission. The objectives of the study were to determine the effect of sodium butyrate on the production of ammonia in the cecal contents of laying hens using in vitro gas production study and to elucidate the mechanism behind it. The study consisted of a control (without sodium butyrate), and three experimental groups added with 10, 15, and 20 mg of sodium butyrate, respectively. Results showed that ammonia production in headspace of the syringe decreased by 8.2, 23, and 23 %, respectively, while ammonium production from the fermentation broth decreased by 6.3, 14.4, and 13.7 %, respectively. Sodium butyrate had no significant effect on the contents of uric acid and urea, nitrate-N, or total N in all treatments. However, sodium butyrate decreased the urease and uricase activities (P
Fulltext Resistance Switching Behavior in Rectangle-Nano-Pattern SrTiO3 Induced by Simple Annealing

Liao X, Zhang Y, Wang J, Kang J, Zhang J, Wang J, et al.

Materials (Basel), 2019 Nov 09;12(22).
PMID: 31717524 DOI: 10.3390/ma12223698

The tunability of semi-conductivity in SrTiO3 single crystal substrates has been realized by a simple encapsulated annealing method under argon atmosphere. This high temperature annealing-induced property changes are characterized by the transmission spectra, scanning electron microscopy (SEM) and synchrotron-based X-ray absorption (XAS). We find the optical property is strongly influenced by the annealing time (with significant decrease of transmittance). A sub gap absorption at ~427 nm is detected which is attributed to the introduction of oxygen vacancy. Interestingly, in the SEM images, annealing-induced regularly rectangle nano-patterns are directly observed which is contributed to the conducting filaments. The XAS of O K-edge spectra shows the changes of electronic structure by annealing. Very importantly, resistance switching response is displayed in the annealed SrTiO3 single crystal. This suggests a possible simplified route to tune the conductivity of SrTiO3 and further develop novel resistance switching materials.
Effects of different types of biochar on methane and ammonia mitigation during layer manure composting

Chen W, Liao X, Wu Y, Liang JB, Mi J, Huang J, et al.

Waste Manag, 2017 Mar;61:506-515.
PMID: 28117129 DOI: 10.1016/j.wasman.2017.01.014

Biochar, because of its unique physiochemical properties and sorption capacity, may be an ideal amendment in reducing gaseous emissions during composting process but there has been little information on the potential effects of different types of biochar on undesired gaseous emissions. The objective of this study was to examine the ability and mechanism of different types of biochar, as co-substrate, in mitigating gaseous emission from composting of layer hen manure. The study was conducted in small-scale laboratory composters with the addition of 10% of one of the following biochars: cornstalk biochar, bamboo biochar, woody biochar, layer manure biochar and coir biochar. The results showed that the cumulative NH3 production was significantly reduced by 24.8±2.9, 9.2±1.3, 20.1±2.6, 14.2±1.6, 11.8±1.7% (corrected for initial total N) in the cornstalk biochar, bamboo biochar, woody biochar, layer manure biochar and coir biochar treatments, respectively, compared to the control. Total CH4 emissions was significantly reduced by 26.1±2.3, 15.5±2.1, 22.4±3.1, 17.1±2.1% (corrected for the initial total carbon) for cornstalk biochar, bamboo biochar, woody biochar and coir biochar treatments than the control. Moreover, addition of cornstalk biochar increased the temperature and NO3(-)-N concentration and decreased the pH, NH4(+)-N and organic matter content throughout the composting process. The results suggested that total volatilization of NH3 and CH4 in cornstalk biochar treatment was lower than the other treatments; which could be due to (i) decrease of pH and higher nitrification, (ii) high sorption capacity for gases and their precursors, such as ammonium nitrogen from composting mixtures, because of the higher surface area, pore volumes, total acidic functional groups and CEC of cornstalk biochar.
Sodium butyrate reduces ammonia emissions through glutamate metabolic pathways in cecal microorganisms of laying hens

Zhong Z, Wang C, Zhang H, Mi J, Liang JB, Liao X, et al.

Ecotoxicol Environ Saf, 2022 Feb 14;233:113299.
PMID: 35176673 DOI: 10.1016/j.ecoenv.2022.113299

Ammonia emission is an important problem that needs to be solved in laying hen industries. Sodium butyrate (SB) is considered to have potential for reducing ammonia production because of its ability to improve nitrogen metabolism. In this in vitro fermentation study, we presented a correlation analysis of the metatranscriptome and metaproteome of lay hen cecal microorganisms, in order to identify important proteins and pathways involved in ammonia production reduction due to sodium butyrate supplementation. The results showed that sodium butyrate supplement decreased the production of ammonia by 26.22% as compared with the non-sodium butyrate supplementation (CK) group. The SB group exhibited a lower concentration of ammonium nitrogen (NH4+-N) and a decreased pH. Sodium butyrate promoted the uric acid concentration and lowered the uricase activity in the fermentation broth of laying hens cecal content. Notably, the 'alanine, aspartate and glutamate metabolism' category was more abundant in the SB group. The addition of sodium butyrate increased the expression of glutamate dehydrogenase (GDH) gene in cecal microbiota (e.g., Ruminococcus sp. and Bacteroides sp.) in vitro. The metaproteome analysis results showed that the expression of GDH with NADPH as coenzyme (NADPH-GDH) was up-regulated in cecal microbiota by sodium butyrate supplement. Our results indicate that sodium butyrate can affect glutamate metabolism through regulating the expression of glutamate dehydrogenase in cecal microorganisms, thereby reducing ammonia production. This study reveals that glutamate dehydrogenase-mediated glutamate metabolism play a key role in ammonia emission reduction in laying hen and provide theoretical basis for further developing ammonia production reduction approach.
Effect of the chlortetracycline addition method on methane production from the anaerobic digestion of swine wastewater

Huang L, Wen X, Wang Y, Zou Y, Ma B, Liao X, et al.

J Environ Sci (China), 2014 Oct 1;26(10):2001-6.
PMID: 25288543 DOI: 10.1016/j.jes.2014.07.012

Effects of antibiotic residues on methane production in anaerobic digestion are commonly studied using the following two antibiotic addition methods: (1) adding manure from animals that consume a diet containing antibiotics, and (2) adding antibiotic-free animal manure spiked with antibiotics. This study used chlortetracycline (CTC) as a model antibiotic to examine the effects of the antibiotic addition method on methane production in anaerobic digestion under two different swine wastewater concentrations (0.55 and 0.22mg CTC/g dry manure). The results showed that CTC degradation rate in which manure was directly added at 0.55mg CTC/g (HSPIKE treatment) was lower than the control values and the rest of the treatment groups. Methane production from the HSPIKE treatment was reduced (p<0.05) by 12% during the whole experimental period and 15% during the first 7days. The treatments had no significant effect on the pH and chemical oxygen demand value of the digesters, and the total nitrogen of the 0.55mg CTC/kg manure collected from mediated swine was significantly higher than the other values. Therefore, different methane production under different antibiotic addition methods might be explained by the microbial activity and the concentrations of antibiotic intermediate products and metabolites. Because the primary entry route of veterinary antibiotics into an anaerobic digester is by contaminated animal manure, the most appropriate method for studying antibiotic residue effects on methane production may be using manure from animals that are given a particular antibiotic, rather than adding the antibiotic directly to the anaerobic digester.
Fulltext Different Concentrations of Doxycycline in Swine Manure Affect the Microbiome and Degradation of Doxycycline Residue in Soil

Yan Q, Li X, Ma B, Zou Y, Wang Y, Liao X, et al.

Front Microbiol, 2018;9:3129.
PMID: 30619199 DOI: 10.3389/fmicb.2018.03129

Antibiotic residues that enter the soil through swine manure could disturb the number, community structure and functions of microbiota which could also degrade antibiotics in soil. Five different concentrations of doxycycline (DOX) incorporated into swine manure were added to soil to explore the effects of DOX on microbiota in soil and degradation itself. The results showed that the soil microbiome evolved an adaptation to the soil containing DOX by generating resistance genes. Moreover, some of the organisms within the soil microbiome played crucial roles in the degradation of DOX. The average degradation half-life of DOX in non-sterile groups was 13.85 ± 0.45 days, which was significantly shorter than the 29.26 ± 0.98 days in the group with sterilized soil (P < 0.01), indicating that the soil microbiome promoted DOX degradation. DOX addition affected the number of tetracycline resistance genes, depending on the type of gene and the DOX concentration. Among these genes, tetA, tetM, tetW, and tetX had significantly higher copy numbers when the concentration of DOX was higher. In contrast, a lower concentration of DOX had an inhibitory effect on tetG. At the same time, the microbial compositions were affected by the initial concentration of DOX and the different experimental periods. The soil chemical indicators also affected the microbial diversity changes, mainly because some microorganisms could survive in adversity and become dominant bacterial groups, such as the genera Vagococcus and Enterococcus (which were associated with electrical conductivity) and Caldicoprobacter spp. (which were positively correlated with pH). Our study mainly revealed soil microbiota and DOX degradation answered differently under variable concentrations of DOX mixed with swine manure in soil.
Metabonomics reveals an alleviation of fitness cost in resistant E. coli competing against susceptible E. coli at sub-MIC doxycycline

Wen X, Cao J, Mi J, Huang J, Liang J, Wang Y, et al.

J Hazard Mater, 2021 03 05;405:124215.
PMID: 33109407 DOI: 10.1016/j.jhazmat.2020.124215

High concentrations of antibiotics may induce bacterial resistance mutations and further lead to fitness costs by reducing growth of resistant bacteria. However, antibiotic concentrations faced by bacteria are usually low in common environments, which leads to questions about how resistant bacteria with fitness costs regulate metabolism to coexist or compete with susceptible bacteria during sublethal challenge. Our study revealed that a low proportion (< 15%) of resistant bacteria coexisted with susceptible bacteria due to the fitness cost without doxycycline. However, the cost for the resistant strain decreased at a doxycycline concentration of 1 mg/L and even disappeared when the doxycycline concentration was 2 mg/L. Metabonomics analysis revealed that bypass carbon metabolism and biosynthesis of secondary metabolites were the primary metabolic pathways enriching various upregulated metabolites in resistant bacteria without doxycycline. Moreover, the alleviation of fitness cost for resistant bacteria competed with susceptible bacteria at 1 mg/L doxycycline was correlated with the downregulation of the biomarkers pyruvate and pilocarpine. Our study offered new insight into the metabolic mechanisms by which the fitness cost of resistant mutants was reduced at doxycycline concentrations as low as 1 mg/L and identified various potential metabolites to limit the spread of antimicrobial resistance in the environment.
Fulltext Changes in the Carbon Metabolism of Escherichia coli During the Evolution of Doxycycline Resistance

Yang Y, Mi J, Liang J, Liao X, Ma B, Zou Y, et al.

Front Microbiol, 2019;10:2506.
PMID: 31736928 DOI: 10.3389/fmicb.2019.02506

Despite our continuous improvement in understanding the evolution of antibiotic resistance, the changes in the carbon metabolism during the evolution of antibiotic resistance remains unclear. To investigate the evolution of antibiotic resistance and the changes in carbon metabolism under antibiotic pressure, Escherichia coli K-12 was evolved for 38 passages under a concentration gradient of doxycycline (DOX). The 0th-passage sensitive strain W0, the 20th-passage moderately resistant strain M20, and the 38th-passage highly resistant strain E38 were selected for the determination of biofilm formation, colony area, and carbon metabolism levels, as well as genome and transcriptome sequencing. The MIC of DOX with E. coli significantly increased from 4 to 96 μg/ml, and the IC50 increased from 2.18 ± 0.08 to 64.79 ± 0.75 μg/ml after 38 passages of domestication. Compared with the sensitive strain W0, the biofilm formation amount of the resistant strains M20 and E38 was significantly increased (p < 0.05). Single-nucleotide polymorphisms (SNPs) were distributed in antibiotic resistance-related genes such as ribosome targets, cell membranes, and multiple efflux pumps. In addition, there were no mutated genes related to carbon metabolism. However, the genes involved in the biosynthesis of secondary metabolites and carbon metabolism pathway were downregulated, showing a significant decrease in the metabolic intensity of 23 carbon sources (p < 0.05). The results presented here show that there may be a correlation between the evolution of E. coli DOX resistance and the decrease of carbon metabolism, and the mechanism was worthy of further research, providing a theoretical basis for the prevention and control of microbial resistance.
Influence and characteristics of Bacillus stearothermophilus in ammonia reduction during layer manure composting

Wang Y, Bi L, Liao Y, Lu D, Zhang H, Liao X, et al.

Ecotoxicol Environ Saf, 2019 Sep 30;180:80-87.
PMID: 31078019 DOI: 10.1016/j.ecoenv.2019.04.066

Ammonia emissions is an important issue during composting because it can cause secondary pollution and a significant of nitrogen loss. Based on research adding Bacillus stearothermophilus can reduce ammonia emissions during composting because it can use sugar in organic matter fermentation to produce organic acids over 50 °C. This study conducted the batch experiments by adding different concentrations of Bacillus stearothermophilus to reduce the ammonia emissions and find out its characteristic during layer manure composting by using an aerobic composting reactor with sawdust as a bulking agent. The results show that the application of Bacillus stearothermophilus can accelerate the rate of temperature and significantly decrease pH, the warming period was 2 days in the treatment with Bacillus stearothermophilus, while it was 4 days in the treatment without Bacillus stearothermophilus. Ammonia emissions were mainly occurred in warming and high temperature period during composting. The ammonia emissions in the treatment with 8.00 g/kg initial Bacillus stearothermophilus were significantly lower than the other lower Bacillus stearothermophilus treatment and control during composting (p  0.05). MiSeq System Sequencing results find that the addition of Bacillus stearothermophilus changed the bacterial community structure under warming and high-temperature periods during composting, increased the relative abundance of lactic acid bacillus and nitrification bacteria. Therefore, the reason for the low ammonia emission in 8.00 g/kg initial Bacillus stearothermophilus treatments might be not only due to the Bacillus stearothermophilus itself, but also Bacillus stearothermophilus can change the indigenous microorganism community, including increase the relative content of lactic acid Bacillus and nitrification bacteria, thus reducing the pH and promoting nitrification, and reducing ammonia emissions.
Heterologous expression of the tetracycline resistance gene tetX to enhance degradability and safety in doxycycline degradation

Wen X, Huang J, Cao J, Xu J, Mi J, Wang Y, et al.

Ecotoxicol Environ Saf, 2020 Mar 15;191:110214.
PMID: 31968275 DOI: 10.1016/j.ecoenv.2020.110214

Microbial remediation has the potential to inexpensively yet effectively decontaminate and restore contaminated environments, but the virulence of pathogens and risk of resistance gene transmission by microorganisms during antibiotic removal often limit its implementation. Here, a cloned tetX gene with clear evolutionary history was expressed to explore doxycycline (DOX) degradation and resistance variation during the degradation process. Phylogenetic analysis of tetX genes showed high similarity with those of pathogenic bacteria, such as Riemerella sp. and Acinetobacter sp. Successful tetX expression was performed in Escherichia coli and confirmed by SDS-PAGE and Western blot. Our results showed that 95.0 ± 1.0% of the DOX (50 mg/L) was degraded by the recombinant strain (ETD-1 with tetX) within 48 h, which was significantly higher than that for the control (38.9 ± 8.7%) and the empty plasmid bacteria (8.8 ± 5.1%) (P 0.05). The efficient and safe DOX-degrading capacity of the recombinant strain ETD-1 makes it valuable and promising for antibiotic removal in the environment.
Occurrence and contamination profiles of antibiotic resistance genes from swine manure to receiving environments in Guangdong Province southern China

Wen X, Mi J, Wang Y, Ma B, Zou Y, Liao X, et al.

Ecotoxicol Environ Saf, 2019 May 30;173:96-102.
PMID: 30769208 DOI: 10.1016/j.ecoenv.2019.02.023

Livestock farms are commonly regarded as the main sources of antibiotic resistance genes (ARGs), emerging pollutants with potential implications for human health, in the environment. This study investigated the occurrence and contamination profiles of nine ARGs of three types from swine manure to receiving environments (soil and water) in Guangdong Province, southern China. All ARGs occurred in 100% of swine manure samples. Moreover, the absolute concentration of total ARGs varied from 3.01 × 108 to 7.18 × 1014 copies/g, which was significantly higher than that in wastewater and manured soil (p  0.05). However, the number of ARGs (ermB, qnrS, acc(6')-Ib, tetM, tetO and tetQ) decreased but were not eliminated by wastewater treatment components (p

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