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

that the ratio of Bacillus groups increased to 54.4% and the bacterial number decreased after the thermophilic phase (Watanabe et al., 2015b), which were similar to those of the other reports (Cahyani et al., 2003; Partanen et al., 2010; Rebollido et al., 2008; Sasaki et al., 2009; Schloss et al., 2005; Yamamoto et al., 2009), the higher ratio of gram-positive bacterial groups seemed be caused by a higher survival ratio of relatively thermotolerant gram-positive bacterial groups during the thermophilic phase (Roman et al.,2015). In contrast, there was no numerically dominant bacterial group in composts originating from cattle feces (E, F1, F2, and F3), and the ratios of gram-positive bacterial groups became lower (22.5% to 57.4%) than those of the former (A, B, BB, C, and D) (Table 1, Figure 1). This difference might be caused from the lower maximum temperature attained during the thermophilic phase, which was not enough to eliminate fecal bacteria and increase thermotolerant bacterial groups (Roman et al.,2015). As compost F1, F2, and F3 was made from the same cattle feces with the same composting process, differences in bacterial composition were caused from the difference in thermophilic condition, which was resulted from difference in starting condition as described in Material and Method (Table 1, Figure 1). As typical fecal bacteria and pathogenic bacteria was detected in the tested composts, such as Clostridium perfringens (C.perfri50, C.perfring, CP000246 ,M59103), Fusobacterium nucleatum (Fus.nuclea, AE009951, AJ133496) or F.sunuae (Fus.simiae) in compost B, Clostridium botulinum (L37585, L37587, C.botulin6), Mycoplasma salivarium (M.salivari), and Prevotellaoris (L16474) or Bacteroides eggerthii (L16485) in compost BB, Bacteroides sp. (AY008308), Clostridium butyricum (AY442812, C.butyric2, C.butyric3, C.butyric4), and Fusobacterium sp. (AF287805, AF385575, AF432130) in compost C, Ehrlichiasp. (Ehr.ris081, Ehr.risKEN, Her.ristic, M73225) or E.sennetsu (M73225), and Leptonema illini (Lpn.illini,Z21632) in compost D, Bordetella sp. (DQ132877) and Fusobacterium nucleatum (Fus.nuclea) in compost E, and Fusobacterium nucleatum (Fus.nuclea) and Parachlamydia sp. (AF366365, AJ715410) or Spirillum winogradskii (AY845251) in compost F3 (Table S1), these composts were indicated to include bacteria of fecal origin. As typical fecal bacteria such as Fusobacterium sp., Borrelia anserine, and Leptospira fainei, were also detected in the former studies (Watanabe et al.2015b), the fecal bacteria and pathogenic bacteria were not always completely eliminated during the composting process, as reported in other results (Brinton et al., 2009; Reynnells et al., 2014). As typical fecal bacteria such as Mycoplasma sualvi (M. sualvi), Prevotellanuminicola (AB003401), P. oralis (L16480), and Spiroplasma sp. (M24662, Spp.cit2HP, Spp.poulsn) had been detected in paddy field soil annually applied with compost (Watanabe et al.,2015a), compost application was suggested to disperse fecal bacteria originating from livestock to field soil. b) Phylogenetic estimation and enumeration of MRB In the composts originating from chicken droppings and pig feces (compost A, B, BB, C, and D), compost C included a considerable number of MRB (1.0 x104MPN g-1), and compost D included a higher number of MRB (84.9 x104MPN g-1) (Table 2). The composts originating from cattle feces (compost E, F1, F2, and F3) included MRB from 43.2 x104MPN to 84.9 x104MPN g-1 (Table 2). There was a large difference between the composition of general bacteria and that of MRB, where the gram-positive bacterial group was not numerically dominant (Figure 1, and 2). Uncultured Sphigomonadaceae (AF408325) was the numerically dominant MRB in compost E (67.1x x104MPN) and various Sphnigomonas sp. were the numerically dominant MRB in compost F1 (Table 2, and Figure 2). As composts F1, F2, and F3 were made from the same cattle feces under the same composting process, the compositional difference of MRB among the composts was suggested to be caused from a slight difference in starting conditions (Table 2, Figure 2). As typical fecal bacteria, such as Bacteroides coprocola (AB200223, AB200225, AB200225) and Borrelia recurrent is(AF107356, U42300), were detected as MRB in compost E, and Bacteroides bacterium (AY162121) was detected in compost F2 (Table 2S), these composts were indicated to include MRB of fecal origin. The present data indicated that most composts used by organic farmers in various region of Japan not only included MRB but also pathogenic bacteria of livestock origin. The present results were enough to promote awareness that these hazardous bacteria might contaminate fresh vegetables from field soils, as suggested by the other reports (Watanabe 2008, 2009; Watanabe et al.,2015a; Yong et al., 2016).As elimination of contaminated hazardous bacteria from field soils was difficult (Watanabe 2008, 2009; Watanabe et al.,2015a), their existence in compost had to be checked before spreading into field soil. For this purpose, the method used in this manuscript was found to be suitable. IV. C onclusions Composting is a biological aerobic decomposition process of biomaterials consisting of two different phases: first, the thermophilic phase and next, the maturing phase (Misra et al., 2003; Roman et al., 2015). First, in the thermophilic phase, microbiological degradation of easily degradable biomaterial in feces elevated the temperatures and diminished the moisture content, while fecal bacteria in livestock feces were eliminated and only thermotolerant bacterial groups 22 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