HM Medical Clinic

J. Viet. Env. 2014, Vol. 6, No. 1, pp. 77-83
DOI: 10.13141/jve.vol6.no1.pp77-83
Identification of antibiotic-producing Bacillus
sensu lato
isolated from national parks of
Hoang Lien and Phu Quoc in Vietnam

Phân loại các loài vi khuẩn Bacillus sensu lato sinh kháng sinh phân lập tại vườn Quốc Gia Hoàng Liên và Phú Quốc 1 Department of Vietnam Type Culture Collection, Institute of Microbiology and Biotechnology, Vietnam National Uni-versity, Hanoi, 144-Xuan Thuy-Cau Giay, Hanoi, Vietnam; 2Department of Medical Microbiology, Bach Mai hospital, 76-Giai Phong-Dong Da, Hanoi, Vietnam Many lipopeptide antibiotics produced from Bacillus sensu lato (Bacillus s. l.) against drug-resistant bacteria have been recently reported. To explore the potential production of the antibiot-ics from this group of bacteria in Vietnam, we collected 38 soil samples from two national parks of Hoang Lien and Phu Quoc and isolated 411Bacillus s. l. strains. Of those, 22 strains had antag-onistic activity against both susceptible S. aureus and E. coli. The strains were further tested on drug-resistant bacteria collected at Bach Mai hospital and 20 strains demonstrated antagonistic ac-tivity against at least 2 of 18 drug-resistant bacteria K. pneumoniae, E. coli, A. baumannii and S. aureus. Analysis of 16S rDNA sequence showed that most of the broad spectrum antibiotic pro-ducers were Paenibacillus species whereas narrow spectrum antibiotic producers were Bacillus species. Strains PQH 0103 and PQH 0410 were probably produced novel antibiotic agents as they suspected to be novel taxa in Paenibacillus genus. Moreover, strains PQH 1509 and PQH 1702 produced broad spectrum antibiotics were identified as P. chitinolyticus. To our best knowledge, there is no report on antibiotic production from this species. Further elucidation of chemical struc-ture of antibiotics produced from Bacillus s. l. isolated in soils of Vietnam is needed. Gần đây, nhiều kháng sinh lipopeptide từ vi khuẩn Bacillus sensu lato (Bacillus s. l.) có khả năng sinh kháng sinh kháng vi khuẩn kháng thuốc đã được phát hiện và công bố. Để tìm hiểu khả năng sinh kháng sinh của nhóm vi khuẩn này ở Việt Nam, chúng tôi tiến hành thu thập 38 mẫu đất tại vườn Quốc gia Hoàng Liên và vườn Quốc gia Phú Quốc và phân lập được 411 chủng vi khuẩn Bacillus s. l.. Trong số đó, 22 chủng có khả năng sinh kháng sinh kháng 2 chủng vi khuẩn S. aure-usE. coli nhậy cảm thuốc. 20 trong số 22 chủng này có hoạt tính kháng ít nhất từ 2 đến 18 chủng kiểm định kháng thuốc thuộc các loài K. pneumoniae, E. coli, A. baumanniiS. aureus. Dựa trên kết quả phân loại bằng trình tự gene 16S rRNA, các chủng Bacillus s. l. sinh kháng sinh phổ rộng là các loài Paenibacillus trong khi đó các loài sinh kháng sinh phổ hẹp là Bacillus. Trên cây phát sinh chủng loại, chủng PQH 0103 và PQH 0410 có thể là những loài mới và có tiềm năng sinh các chất kháng sinh mới. Hơn nữa, chủng PQH 1509 và PQH 1702 sinh kháng sinh phổ rộng được phân loại vào loài P. chitinolyticus. Đây là loài chưa có công bố nào về khả năng sinh chất kháng sinh và có thể là loài sinh chất kháng sinh mới. Nghiên cứu giải mã cấu trúc chất kháng sinh từ các loài Bacillus s. l. phân lập tại Việt Nam cần được làm rõ trong tương lai. Keywords: antibiotic, antimicrobial agent, Bacillus sensu lato, biodiversity, drug-resistant
bacteria and identification.
* Corresponding author E-mail: [email protected]
J. Viet. Env. 2014, Vol. 6, No. 1, pp. 77-83
1. Introduction
formed under biological safety cabinet class II in a re- Since the discovery of penicillin, hundreds of antibiotics stricted area of the Institute. originated from microorganisms have been discovered, developed and used in medical clinics for treating infec- 2.2 Soil sampling and bacterial isolation
tious diseases (Walsh, 2003; Mandell et al., 2005). How- ever, pathogenic bacteria exposed to antibiotic-containing Thirty-eight soil samples were collected at a depth of 10 environments can mutate their DNA content or acquire cm at two national parks of Hoang Lien (18 samples) and resistant genes by the mean of horizontal gene transfer in Phu Quoc (20 samples) in November, 2010 and 2011, order to develop their own resistant mechanisms which respectively. For each sample, 10 grams of soil were result in the formation of drug-resistant bacteria (Toeno- dispersed into a 250 ml Erlenmeyer flask containing 90 ver, 2006; Alekshun and Levy, 2007). ml of NaCl 0.85%. After shaking at 220 rpm for 30 min, the bacterial supernatants was collected and heated at Emergence of drug-resistant bacteria is major global 80oC for 10 min in order to eliminate vegetable cells. The health threat associated with high mortality rates and treated supernatant was then diluted and plated into Nu- medical costs. In a recent joint technical report, the Euro- trient agar (Becton Dickinson and Company, France). pean Medicines Agency (EMA) in collaboration with After incubation at 25oC for 5 days, colonies demonstrat- Action on Antibiotic Resistance (ReAct) estimated that at ed distinct characteristics of size, colour, surface texture least 25,000 patients die each year in the EU from an and margin were collected and stored at -70oC in LB infection due to drug-resistant bacteria (Freire-Moran et broth containing 20% glycerol. al., 2011). In USA, the death occurs in one of five patients infected with methicillin-resistant Staphylococcus aureus 2.3 Antagonistic activity test
(MRSA) and there are more death arising from invasive MRSA infection than from AIDS patients (Dougherty and From fresh cultures grown on nutrient agar, Bacillus s. l. strains were streaked on TSB agar (Becton Dickinson and Company, France) and incubated at 28oC for 28 h. Using Although antibiotic resistance occurs in many bacteria sterile steel roll with a diameter of 6 mm, agar plugs were species, Enterococcus faecalis, Staphylococcus aureus, prepared and transferred onto nutrient agar inoculated Klebsiella pneumonia, Acinetobacter baumanni, Pseudo- with certain bacterial indicators. After overnight incuba- monas aeruginosa and Escherichia coli (or the ESKAPE tion at 35oC, a clear inhibition zone surrounding agar bacteria) are the most considerable group of drug-resistant plugs was reported as positive result. bacteria (Shlaes, 2010). This group possesses a high risk of hospital acquired infection and can relate to outbreaks 2.4 16S rDNA sequencing analysis
of diseases (D'Souza et al., 2010; Taneja et al., 2010). Thus, discovery of new antibiotics exhibiting the activity Bacterial DNA was extracted by soft lysis method. Brief- against this bacterial group is crucial needed. ly, fresh bacterial cells were treated in 0.2 ml of lysis buffer (100 mM Tris HCl, 100mM Na2EDTA, 1.5 M Recently, many lipopeptide antibiotics produced from NaCl, 1% cetyltrimethyl ammonium bromide (CTAB), Bacillus sensu lato (Bacillus s. l.) have been reported (He pH 8.0) added with 50 µl of SDS 20%. After mixing vig- et al., 2007; Wu et al., 2010; Guo et al., 2012). This group orously for 20 sec, the cells were incubated at 65oC for 2 of antibiotics shows highly effectiveness against drug- h. An equal volume of chloroform : isoamyl alcohol (49 : resistant bacteria in vitro and invivo (Ding et al., 2011; 1) was added to precipitate protein by a centrifuge step at Huang and Yousef, 2014). In order to explore the antibi- 16,000 g for 5 min. The upper layer of DNA solution was otics from microbial resource in Vietnam, we isolated and transferred into a new tube, precipitated with cooled iso- identified antibiotic-producing Bacillus s. l. in two na- propanol, and washed with cooled ethanol 70%. The tional parks of Hoang Lien and Phu Quoc. DNA pellet was suspended into TE buffer and stored at - 20oC for further analysis. 2. Materials and methods
For 16S rRNA gene amplification, approximately 50 ng 2.1 Bacterial indicators
of template DNA was incorporated into 25-µl reactions containing 1x PCR buffer, 1U Taq polymerase (KaTaRa), Two common susceptible bacteria of Escherichia coli 125 nM of each dNTP, and 400 nM of each universal VTCC-B-482 and Staphylococcus aureus VTCC-B-658 primers 27F and 1525R. The PCR conditions were 95oC available at Vietnam Type Culture Collection were in- for 5 min, following by 35 cycles of 30 sec at 95oC, 30 volved into primary screening test. Additionally, 18 drug- sec at 55oC and 1min 45 sec at 72oC. The PCR products resistant bacteria of S. aureus (3 strains), E. coli (6 were confirmed by gel electrophoresis and then purified strains), K. pneumoniae (6 strains) and A. baumannii (3 using a commercial DNA extraction kit (Qiagen). Nucleo- strains) were kindly received from Department of Medical tide sequences were analyzed with universal primers Microbiology, Bach Mai hospital. Antibiotic susceptibil- 1492R, 800R and 518F on 3100 Avant Genetic Analyzer ity of the bacteria was tested using disk diffusion method. (Applied Biosystem). Quality of nucleotide chromato- The resistance profiles were interpreted using documents gram was checked and edited with Chromas lite 2.1. guided by Clinical Laboratory and Standard Institute Nearly complete 16S rRNA gene sequences ( 1.5 kb) (CLSI). All of viable drug-resistant bacteria were per- were assembled using Clone Manager 8.0.
J. Viet. Env. 2014, Vol. 6, No. 1, pp. 77-83
2.5 Phylogenetic analysis
on the antibiotic susceptibility profiles, 3 strains of S. aureus were sensitive to phenicol, glycopeptide and oxa- All of the 16S rDNA nucleotide sequences were retrieved zolidinone but resistant to penicillins, second generation from GenBank. The sequences were aligned using Clustal of cephalosporin, amiloglycoside, fluoroquinolone, W. Phylogenetic tree was constructed by neighbour- fosfomycin and macrolide; 6 strains of E. coli and 6 joining method using MEGA 5. strains of K. pneumoniae were completely resistant to penicillins, all generations of cephalosporin, fluoroquino- 3. Results and discussion
lone and fosfomycin; 3 strains of A. baumannii were resistant to penicillins, all generations of cephalosporin, amiloglycoside, fluoroquinolone, fosfomycin and poly- 3.1 Isolation of Bacillus sensu lato
myxin. As the pathogenic bacteria resistant to antibiotics in more than three antimicrobial categories, they were Bacillus s. l. is a large and diverse group of endospore- considerable to be multidrug-resistant bacteria (Magio- forming bacilli that occupies in various natural habitats. rakos et al., 2011). This group consists of at least 37 genera belonging to three families Bacillaceae, Paenibacillaceae and Alicy-
clobacillaceae (Goldman and Green, 2009). Based on
characteristic of heat-resistant endospores, vegetative
bacterial cells of heterotrophic soil bacteria were elimi-
nated and the isolation of endspores was performed. To-
tally, 411 Bacillus s. l. strains were isolated from 38 soil
samples collected at Hoang Lien and Phu Quoc national
3.2 Screening for antibiotic-producing
Bacillus sensu lato

3.2.1 Primary screening
Figure 2. Antagonistic pattern of 20 Bacillus sensu lato
strains against drug-resistant bacteria
Primary screening for antibiotic-producing Bacillus s. l. was carried out using two susceptible S. aureus and E. Of 22 Bacillus s. l. strains selected for further test, 9 coli available at VTCC. Of 411 Bacillus s. l. strains, 66 strains produced antibiotics against 4 species of drug- (16%) strains demonstrated antagonistic activity against resistant bacteria K. pneumoniae, E. coli, A. baumannii one or two bacterial indicators. Of those, 17 strains had and S. aureus; 3 strains had antagonistic activity against 3 activity against only Gram-positive S. aureus; 27 strains species of drug-resistant bacteria K. pneumoniae, E. coli against only Gram-negative E. coli; and 22 strains against and S. aureus; 8 had antagonistic activity against only both Gram-positive S. aureus and Gram-negative E. coli drug-resistant S. aureus; and the other 2 strains did not show antibiotic activity (Figure 2). Based on antagonistic pattern, the first 12 strains were classed into broad spec-
trum antibiotic producer and the latter 8 strains were
classed into narrow spectrum antibiotic producers (Figure
2). Of broad spectrum antibiotic producers, 4 strains PQH
0103, PQH 0113, SPH 0904 and SPH 0603 had antibiot-
ics against all of the tested drug-resistant bacteria.

3.3 Identification of antibiotic-producing
Bacillus sensu lato

16S rDNA nucleotide sequences of 22 Bacillus s. l.
strains were analyzed. Based on the closest match with
type strains in the Eztaxon-e database, the bacterial strains
were classified into 4 genera Bacillus (7 strains), Paeni-
(13 strains), Tumebacillus (1 strain), and Viridi-
bacillus (1 strain). In Bacillus, the bacteria were identified Figure 1. Number of Bacillus sensu lato strains
as B. aryabhattai (1 strain), B. cereus (3 strains), B. pu- produce antibiotics against susceptible S. aureus and
milus (1 strain), B. safensis (1 strain) and B. vireti (1 E. coli.
strain). Paenibacillus had P. elgii (2 strains), P. jamilae (3 strains), P. chytinolyticus (2 strains), P. peoriae (1 strain), 3.2.2 Secondary screening
P. terrae (4 strains) and P. chibensis (1 strain). Only one species obtained in each of the two last genera was T. Antibiotic production was further tested on 18 bacteria S. permanentifrigoris (1 strain) and V. arenosi (1 strain). aureus, K. pneumoniae, E. coli and A. baumannii. Based
J. Viet. Env. 2014, Vol. 6, No. 1, pp. 77-83
Totally, 13 species were identified from of 22 Bacillus s.
nibacillus and Tumebacillus genera. Interested in the l. strains (Table 1). broad spectrum antibiotic producers, the phylogenetic tree of Paenibacillus species was constructed. Strain PQH Most of the Bacillus s. l. strains had the 16S rDNA nucle- 0103 and P. tianmuensis formed a separate clade with otide similarity greater than 99.3%. However, 4 strains other closely related species P. elgii, P. koreensis and P. designated as PQH 0103, PQH 0410, PQH 1601 and PQH ehimensis; strain PQH 0410 also located distantly from P. 1005 showed a low 16S rDNA similarity value which was chibensis and others (Figure 3). The result confirmed two in a range from 97.8% to 98.6% (Table 1). All of the potential novel species of Paenibacillus genus. Further strains were isolated from soils of Phu Quoc national assignment of those strains to species level is needed. parks and probably presented novel taxa in Bacillus, Pae-

Table 1. Identification of antibiotic-producing Bacillus sensu lato
based on Eztaxon-e database
No. Strains
Type strain (accession no.)
similarity (%)
enibaci llus elgii SD 17 (AY090110) Pa e n i b a c i llus elgii SD 17 (AY090110) n i cillus jamilae CECT 5266 (AJ271157) n i cillus jamilae CECT 5266 (AJ271157) Paenibaci llus jamilae CECT 5266 (AJ271157) Paenibacillus chytinolyticus IFO 15660 (AB021183) Pa e n i b a c i llus chytinolyticus IFO 15660 (AB021183) n i cillus peoriae DSM 8320 (AJ320494) SPH 0410 Paenibacillus terrae AM141 (AF391124) SPH 0808 Paenibacillus terrae AM141 (AF391124) SPH 0204 Paenibacillus terrae AM141 (AF391124) Paenibacillus terrae AM141 (AF391124) Pa e n i b a c i llus chibensis JCM 9*905 (AB073194) Ba c i l l u s safensis FO-036b (AF234854) ac i l l vireti LMG 21834 (AJ542509) Ba c i l l u s aryabhattai B8W22 (EF114313) c i l l u s pumilus ATCC7061(ABRX01000007) c i l l u s cereus ATCC 15479 (AE016877) c i l l u s cereus ATCC 15479 (PRJNA 57957) c i l l u s cereus ATCC 15479 (PRJNA 57957) cillus permanentifrigoris Eur 19.5 (DQ444975) cillus arenosi LMG 22166 (AJ627212) * The strains did not show antagonistic activity against drug-resistan tbacteria Except for strain PQH 0410, all of the strains identified as thetic mechanisms, the antimicrobial agents are classified Paenibacillus were grouped into broad spectrum antibi- into two types of ribosomally synthesized peptides and otic producers. Four strains PQH 0103, PQH 0113, SPH nonribosomally synthesized peptides. Ribosomally syn- 0904 and SPH 0603 identified as P. elgii and P. jamilae thesized peptides or bacteriocins can exhibit a relatively produced antimicrobial agents against all drug-resistant narrow range of antimicrobial activity as paenibacillin bacteria. P. chibensis PQH 0410 and Bacillus species produced from P. polymyxa (He et al., 2007), paenibacil- produced antibiotics against only Gram-positive S. aure- lin P and paenibacillin N produced from P. alvei (Anan- us. Although T. permanentifrigoris PQH 1005 and V. daraj et al., 2009), pumilicin 4 produced from B. pumilus arenosi SPH 0408 demonstrated antibiotic activity against (Aunpad and Na-Bangchang, 2007), lichenicidin pro- two susceptible S. aureus and E. coli, none of those could duced from B. licheniformis (Dischinger et al., 2009; inhibit the growth of drug-resistant bacteria. Begley et al., 2009) and haloduracin produced from B. halodurans (Lawton et al., 2006). Our Bacillus strains and Bacillus s. l. species have been known to produce a dozen P. chibensis PQH 0410 showing antagonistic activity of peptide and lipopeptide antibiotics. Based on biosyn-
J. Viet. Env. 2014, Vol. 6, No. 1, pp. 77-83
against only Gram-positive S. aureus probably produced
strains probably produced antibiotics related to polymyx- bacteriocin antibiotics. Nonribosomally synthesized peptides show broader spec- Search for new polymyxin derivatives that are less toxici- tra of activities against bacteria or fungi (Abriouel et al., ty to human and higher efficiency against resistant bacte- 2011; Lee and Kim, 2011). After discovery of penicillin, ria is being received a considerable attention in antibiotic some of lipopeptide antibiotics from Bacillus s. l . have discovery from Bacillus s. l.. Many new lipopetide antibi- been used in medical clinics for treatment of infectious otics are recently discovered and reported from Paeni- diseases; they are gramicidin (from Brebacillus brevis), bacillus species such as pelgipeptins from P. elgii strain bacitracin (from B. subtilis) and polymyxin (from P. pol- B69, cyclic lipopeptide antibiotics PE1 and PE2 from P. ymyxa). Of those, polymyxin is playing an important ehimensis strain B7 and octapeptin B5 from P. tianmuen- agent in antibiotic therapy of Gram-negative resistant sis strain F6-B70 (Ding et al., 2011; Qian et al., 2012; bacteria such as A. baumannii, P. aeruginosa, K. pneu- Huang et al., 2013). In phylogenetic tree, those species moniaE. coli (Urban et al., 2010). In phylogenetic were grouped in the same cluster (Figure 3) but the strain tree, the P. jamilae strains SPH 0603, SPH 0904 and PQH PQH 0103 formed a distinct clade which is expected to 1504 and P. peoriae SPH 1208 were grouped in the same produce novel lipopeptide antibiotic. cluster of P. polymyxa (data not shown). Therefore, the 67 Paenibacillus elgii HOA73 (JQ412069) 87 Paenibacillus elgii SD17 (AY090110) 92 PQH 0113
Paenibacillus elgii B69 (GU321104) Paenibacillus koreensis YC300 (AF130254) Paenibacillus ehimensis B7 (JX282195) 90 Paenibacillus ehimensis IFO 15659 (AB021184) Paenibacillus tianmuensis F6-B70 PQH 0103
Paenibacillus gansuensis B518 (AY839866)
Paenibacillus chitinolyticus IFO 15660 (AB021183) 74 PQH 1702
Paenibacillus cookii LMG 18419 (AJ250317) Paenibacillus relictisesami KB0549 (AB567661) Paenibacillus chibensis NRRL B-142 (D85395) Paenibacillus azoreducens CM1 (AJ272249) Paenibacillus favisporus GMP01 (AY208751) Paenibacillus cineris LMG 18439 (AJ575658) Paenibacillus rhizosphaerae CECAP06 (AY751754) Paenibacillus polymyxa DSM 36 (AJ320493) Figure 3. Phylogenetic tree based on 16S rDNA nucleotide sequences of Paenibacillus species

Antibiotic produced from P. chibensis also published by
onistic activity against both susceptible S. aureus and E. Lorentz et al., 2006. This study showed the strong ability coli. Of those, 20 strains demonstrated antagonistic activi- against fungal pathogens of antibiotic produced from P. ty against at least 2 of 18 drug-resistant bacteria of K. chibensis contributed to enhance biocontrolling potential. pneumoniae, E. coli, A. baumannii and S. aureus. Analy- Specially, to our best knowledge, in six researched spe- sis of 16S rDNA sequence showed that most of the broad cies from Paenibacillus genus, there was no report on spectrum antibiotic producers were Paenibacillus species antibiotic production from P. chytinolyticus. Therefore, whereas narrow spectrum antibiotic producers were Ba- we expected that this species produced a potential novel cillus species. Strains PQH 0103 and PQH 0410 were probably produced novel antibiotic agents as they sus- pected to be novel taxa in Paenibacillus genus. Moreover, 4. Conclusions
strains PQH 1509 and PQH 1702 identified as P. chi- tinolyticus produced broad spectrum antibiotics. To our Of 411 Bacillus sensu lato strains isolated from national best knowledge, there is no report on antibiotic produc- parks of Hoang Lien and Phu Quoc, 22 strains had antag- tion from this species. Further elucidation of chemical
J. Viet. Env. 2014, Vol. 6, No. 1, pp. 77-83
structure of antibiotics produced from the Bacillus s. l.
handbook of microbiology. CRC Press. isolated from soils in Vietnam is needed. [12]! Guo, Y., E. Huang, C. Yuan, L. Zhang and A. Yousef, 2012. Isolation of Paenibacillus sp. strain 5. Acknowledgements
and structural elucidation of its broad-spectrum lipopeptide antibiotic" Appl Environ Microbiol 78 This study was supported by National foundation for Science and Technology Development (Nafosted) under grant number 106.03-2012.19 and a microbial resource [13]! He, Z., D. Kisla, L. Zhang, C. Yuan, K. B. Green- programme from Ministry of Science and Technology. Church and A. E. Yousef 2007. Isolation and identification of a Paenibacillus polymyxa strain that 6. References
coproduces a novel lantibiotic and Appl Environ Microbiol 73(1): 168-178. [1]! Abriouel, H., C. M. Franz, N. Ben Omar and A. [14]! Huang Z., Y. Hu, . Shou and M. Song, 2013. 2011. Diversity and applications of Isolation and partial characterization of cyclic bacteriocins. FEMS Microbiol Rev lipopeptide antibiotics produced from Paenibacillus 35(1): 201- 232. ehimensis B7. BMC Microbiol 13: 87. [2]! Alekshun, M. N. and S. B. Levy, 2007. Molecular [15]! Huang E. and A. Yousef, 2014. Paenibacterin, a mechanisms of antibacterial multidrug resistance. Cell 128(6): 1037-1050. neutralises endotoxins and promotes survival in a murine model of Pseudomonas-induced sepsis. Int J [3]! Anandaraj, B., A. Vellaichamy, M. Kachman, A. Antimicrob Agents 44 (1): 74-77. Selvamanikandan, S. Pegu and V. Murugan, 2009. Co-production of two new peptide antibiotics by a [16]! Lawton, E. M., P. D. Cotter, C. Hill and R. P. Ross, bacterial isolate Paenibacillus alvei NP75. Biochem 2007. Identification of a novel two-peptide Biophys Res Commun 379(2): 179-185. lantibiotic, haloduracin, produced by the alkaliphile Bacillus halodurans C-125. FEMS Microbiol Lett [4]! Aunpad, R. and K. Na-Bangchang, 2007. Pumilicin 4, a novel bacteriocin with anti-MRSA and anti-VRE activity produced by newly isolated bacteria [17]! Lee, H. and H. Y. Kim, 2011. Lantibiotics, class I Bacillus pumilus strain WAPB4. Curr Microbiol bacteriocins from the genus Bacillus. J Microbiol Biotechnol 21(3): 229-235. [5]! Begley M, Cotter PD, Hill C, Ross RP (2009) [18]! Lozrentz R. H., Artico S., Silveira A. B., Einsfeld Identification of a novel two-peptide lantibiotic, A., Corcal G., 2006. Evaluation of antimicrobial ac- lichenicidin, following rational genome mining for tivity in Paenibacillus spp. strains isolated from nat- LanM proteins.Appl Environ Microbio75: 5451- ural environment. Lett in Appl Microbiol 43(2006): [6]! Ding, R., X. C. Wu, C. D. Qian, Y. Teng, O. Li, Z. J. [19]! Mandell, G., R. Dolin, J. Bennett, 2005. Principles Zhan and Y. H. Zhao, 2011. Isolation and and practice of infectious diseases. Churchill identification of lipopeptide antibiotics from Paenibacillus elgii B69 with inhibitory activity [20]! Magiorakos, A. P., A. Srinivasan, R. B. Carey, Y. against methicillin-resistant Staphylococcus aureus. Carmeli, M. E. Falagas, C. G. Giske, S. Harbarth, J. J Microbiol 49 (6): 942-949. F. Hindler, G. Kahlmeter, B. Olsson- Liljequist, D. [7]! Dischinger, J., M. Josten, C. Szekat, H. G. Sahl and L. Paterson, L. B. Rice, J. Stelling, M. J. Struelens, G. Bierbaum, 2009. Production of the novel two- A. Vatopoulos, J. T. Weber and D. L. Monnet, 2011. peptide lantibiotic lichenicidin by Bacillus Multidrug-resistant, extensively drug-resistant and licheniformis DSM 13. PLoS One 4(8): e6788. pandrug-resistant bacteria: an international expert proposal for interim standard definitions for acquired [8]! Dougherty, T. J. and M. J. Pucci, 2011. Antibiotic resistance. Clin Microbiol Infect 18(3): 268-281. discovery and development Springer. [21]! Qian C. D., X. C. Wu, Y. Teng, W. P. Zhao, O. Li, [9]! D'Souza, N., C. Rodrigues and A. Mehta, 2010. S. G. Fang, Z. H. Huang and H. C. Gao, 2012. Molecular characterization of methicillin-resistant Battacin (Octapeptin B5), a new cyclic lipopeptide Staphylococcus aureus with emergence of epidemic antibiotic from Paenibacillus tianmuensis clones of sequence type (ST) 22 and ST 772 in agaisnt multidrug-resistant Gram-negative bacteria. Mumbai, India. J Clin Microbiol 48(5): 1806-1811. Antimicrobial Agent Chemotherapy 56(3): 1458- [10]! Freire-Moran, L., B. Aronsson, C. Manz, I. C. Gy- ssens, A. D. So, D. L. Monnet, O. Cars, 2011. Criti- [22]! Shlaes, D. M., 2010. Antibiotics: The perfect storm. cal shortage of new antibiotics in development Springer Science. against multidrug-resistant bacteria-Time to react is now. Drug Resist Updat 14(2):118-124. [23]! Taneja, J., B. Mishra, A. Thakur, V. Dogra and P. Loomba, 2010. Nosocomial blood-stream infections [11]! Goldman, E. and L. H. Green, 2009. Practical from extended-spectrum-beta-lactamase-producing
J. Viet. Env. 2014, Vol. 6, No. 1, pp. 77-83
Escherichia coli and Klebsiella pneumonia from GB and Escherichia coli. Antimicrob Agents Chemother Pant Hospital, New Delhi" I Infect Dev Ctries 4(8): 54(6): 2732-2734. [26]! Walsh, C., 2003. Antibiotics: actions, origins [24]! Tonover, F. C., 2006. Mechanisms of antimicrobial resistance. ASM press. resistance in bacteria. Am J Med 119 (6A): 3-10. [27]! Wu, X.C, Shen, X.B, Ding, R. , Qian, C.D, Fang, [25]! Urban, C., N. Mariano and J. J. Rahal, 2010. In vitro H.H & Li, O. (2010) "Isolation and partial character- double and triple bactericidal activities of ization of antibiotics produced by Paenibacillus elgii doripenem, polymyxin B, and rifampin against multidrug-resistant Acinetobacter baumannii, Pseudomonas aeruginosa, Klebsiella pneumoniae,


Fcc19 mise en page

SOMMAIRE G.MENTHA (Genève) F.-R. PRUVOT (Lille) J.-Y. MABRUT (Lyon) Kyste biliaire simple et formes compliquées P. PESSAUX (Strasbourg) Polykystose hépatique J. HARDWIGSEN (Marseille) Cystadénome biliaire, cystadénocarcinome et autres tumeurs malignes kystiques O. SCATTON (Paris)

PRODUCT INFORMATION NAME OF THE MEDICINE Granisetron Kabi Concentrated Injection Granisetron hydrochloride has the following chemical structure: Empirical formula: Molecular weight: The systematic chemical name is endo-N-(9-methyl-9-azabicyclo [3.3.1] non-3-yl)-1-methyl-1H-indazole-3-carboxamide hydrochloride. DESCRIPTION Granisetron hydrochloride is a white to off-white crystalline powder which is freely soluble in water and sodium chloride 0.9% at 20°C. Granisetron Kabi Concentrated Injection contains granisetron hydrochloride equivalent to granisetron free base 1 mg/mL. It also contains sodium chloride, citric acid monohydrate, hydrochloric acid, sodium hydroxide and water for injections. PHARMACOLOGY Granisetron is a potent anti-emetic and highly selective antagonist of 5-hydroxytryptamine (5-HT3) receptors. Radioligand binding studies have demonstrated that granisetron has negligible affinity for other receptor types, including 5-HT, alpha1 and alpha2, beta-adrenoreceptors, histamine H1, picrotoxin, benzodiazepine, opioid and dopamine D2 binding sites. Antagonism of 5-HT receptors located peripherally on vagal nerve terminals