Global Journal of Medical Research, K: Interdisciplinary, Volume 22 Issue 3

survived (Cahyani et al., 2003; Partanen et al., 2010; Rebollido et al., 2008; Roman et al., 2015; Sasaki et al., 2009; Schloss et al., 2005; Yamamoto et al., 2009). However, the attained maximum temperature and reduction of moisture content were varied depend on starting conditions and air supply during this phase (Roman et al., 2015), which would affect numbers of residual fecal bacteria. Significant positive correlation of the total bacterial number (2) with those of the gram-positive bacterial group (3) (R=0.976, P<0.001 n=9, Table 3) suggested that variation in number of thermotolerant gram-positive bacteria caused the major bacterial difference among the tested 9 composts, which might be mainly affected by a conditional difference in thermophilic phase and could be used as an index to speculate the condition of the thermophilic phase for each compost. As the ratio of the gram-positive bacterial number to the total bacterial number (4) had significant negative correlation to those of gram-negative MRB (6) (r=-0.723,<0.01, n=9, Table 3), those of the MRB of Sphingomonadacea (7) (r=-0.687,<0.05, n=9), and those of the MRB of the other α -Proteobacteria (8)(r=- 0.901,<0.001, n=9), numbers of most ofthe MRB in the composts varied inreversal trend against that of the gram-positive bacterial group (Oliver et al.,2020; Sharma et al., 2009; Wang et al.,2015; Youngquist et al., 2016). As the ratio of gram-positive bacteria would become higher by an effective thermophilic phase, where a higher maximum temperature and lower moisture content was attained (Misra et al., 2003; Roman et al.,2015), MRB might be reduced by the same abiotic factors during thermophilic phase. Moisture content (1), which had decreased by an effective thermophilic phase (Misra et al., 2003; Roman et al., 2015), had positive correlations with the ratios of MRBs ((5)-(8) from r=0.384 to r=0.791; Table 3). Significant positive correlation between moisture content (1) and the ratio of gram-negative MRB (7) (R=0.791, P<0.05, Table 3) suggested that moisture content might be a critical factor to eliminate gram- negative MRB during a thermophilic phase. The elimination of MRB by controlling the composting process will be presented in the next manuscript. A cknowledgments Part of this research was achieved in the Research Team for Biomass Recycling System, in the National Agriculture and Food Research Organization of Japan, supported by a grant under the theme of “Analyses of heavy metal and antibiotic resistant bacteria in composts,“ funded by Japanese Ministry of Agriculture, Forestry and Fisheries from April, 2008 to March, 2010. The author thanks the organic farmers in various regions of Japan for sending the composts used in this experiment. Thank is also given to Mrs. K. Matsuoka for supporting the experiments. The other part of this research was achieved in Department of Life, Environment and Applied Chemistry, Fukuoka Institute of Technology (FIT) supported by a grant (2010) under the theme of “Development of rapid analysis method for microorganisms in aim to contribute environmental protection and recycling biological resources,” funded by FIT, a grant (2014) under the theme of “Development of new method for identification and quantification of bacteria using microchip electrophoresis” funded by the Japan Science and Technology Agency, and a grant (2018) under the theme of “Practical realization of the method for fact-finding survey of antibiotic resistant bacteria in environment and foods” funded by the economic council for the Kyushu region. The author thank Mr.PatrickSulsar in the Office of International Education in FIT for corrections of grammatical errors in this manuscript. Supplemental Material Supplemental Table 1 shows the phylogenetic estimations of general bacteria in each dilution vial, whose DNA was extracted after the incubation of diluted samples in an LB medium. Abbreviations: ARG antibiotic resistant gene; MERFLP multiple enzyme restriction fragment length polymorphism; MPN most provable number; MRB multidrug resistant bacteria; NGS next generation sequencing; qPCR quantitative polymerase chain reaction; RDP the Ribosomal Database Project. R eferences R éférences R eferencias 1. Agga, G.E., T.M. Arthur, L.M. Durso, D.M. Harhay, & J.W. Schmidt. 2015. Antimicrobial-Resistant Bacterial Populations and Antimicrobial Resistance Genes Obtained from Environments Impacted by Livestock and Municipal Waste. PLoS ONE. 10: e0132586. doi:10.1371/journal.pone.0132586. 2. Blodgett, R. 2010. FDA, Bacterial Analytical Manual, Appendix 2: Most Probable Number from Serial Dilutions. http://www.fda.gov/Food/FoodScience Research/LaboratoryMethods/ucm109656.htm/ 3. Brinton Jr, W.F., P. Storms, & T.C. Blewett. 2009. Occurrence and Levels of Fecal Indicators and Pathogenic Bacteria in Market-Ready Recycled Organic Matter Composts. Journal of Food Protection. 72: 332–339.doi: 10.4315/0362-028x- 72.2.332. 4. Burgos, J.M., B.A. Ellington, & M.F. Varela. 2005. Presence of Multidrug Resistant Enteric Bacteria in Dairy Farm Topsoil. Journal of Dairy Science. 88: 1391-1398. http://dx.doi.org/10.3168/jds.S0022- 0302(05)72806-X. 5. Cahyani, V.R., K. Matsuya, S. Asakawa, & M. Kimura. 2003. Succession and Phylogenetic Composition of Bacterial Communities Responsible 23 Year 2022 Global Journal of Medical Research Volume XXII Issue III Version I ( D ) K © 2022 Global Journals Dispersion of Multidrug Resistant Bacteria and Fecal Bacteria into Field Soils of Japan through Compost Application