We excluded patients who were classified as a housewife or student. The hospitals belong to a nonprofit organization that

HSP inhibitor provides special attention for occupation-related conditions and were founded by the Ministry of Health, Labour and Welfare of Japan. Stroke was diagnosed using the international classification of diseases, 10th revision (ICD-10) codes for cerebral hemorrhage, cerebral infarction, or subarachnoid hemorrhage. Outcome measure The outcome was return to work after stroke, which was defined as active employment in formal paid work on a full-time or part-time basis which was identified at follow-up 18 months after the onset of stroke. The information was reported directly by patients, by physiatrists at the outpatient clinic interviewing patients, or by trained clerical staff interviewing patients by telephone at 18 months after onset. Procedures A unified electronic data format was used to extract patient information from hospital records at the time of admission, discharge, and follow-up 18 months post-stroke. Data were collected on history and lifestyle factors, Selleckchem Midostaurin demographic factors, diagnostic factors, functional factors, and occupational factors. Physiatrists interviewed patients

to obtain information regarding history and lifestyle factors at initial rehabilitation and collected clinical and diagnostic factors at discharge from medical records. Higher cortical dysfunction (brain impairment related to behavior, cognition, and language that cannot be explained by motor paralysis or sensory or perception disorders)

was diagnosed by neurologists using the neurological examination based on higher cortical dysfunction diagnosis guidelines (Japanese Ministry of Health, Labour and Welfare 2007), the Standard Language Test of Aphasia (Japan Society for Higher Brain Dysfunction 2003), the Mini-Mental State Examination (Folstein et al. 1975), the line bisection test, and the Kohs block test. Radiologists independently and in a blinded many manner made diagnoses regarding etiology, anatomical location, and size of stroke by neuroradiological imaging. Occupational therapists evaluated functional factors with the modified Rankin scale (mRS) (van Swieten et al. 1988) and the Barthel index (BI) (Malloney and Barthel 1965). The BI is a measure of functional ability in personal care including self-care, bowel and bladder sphincter control, and mobility. Job type was classified according to the Japanese standard classification of occupations (Japanese Ministry of Health, Labour and Welfare 1997). We classified the following jobs as white collar: clerks, technicians, highly skilled professionals, directors, and managers. Unskilled workers, production-line/machine workers, drivers, skilled manual workers, farm/horticulture workers, and service workers were classified as blue collar.

Another 2 cases with no clinical treatment had a neuroradiological diagnosis of radiation necrosis and were under observation. Figure 1 Typical MRI scan changes in ACTH adenoma. Coronal T1-weighted postcontrast MRI scan at left and right, obtained in Patient 1, a 30-year-old man who presented with ACTH adenomas and consistent headache

2 years before undergoing GKRS. An enhancing mass lesion is seen in the sella turcia with extension to bilateral internal carotid artery. Patient 1′s serum ACTH level was 381.6 pg/ml, and his blood pressure was over 180/120 mmHg. The patient was treated with MASEP GKRS, and MRI was performed for treatment planning. 26 Gy defined to the 50% isodose line is used to cover the full extent of the pituitary tumor in all three planes. Figure 2 Typical MRI scan changes in ACTH adenoma. No enhancing mass lesion is seen in the sella turcia under the T1-weighted postcontrast MRI scan performed 2 years after GKRS. Patient LEE011 cell line 1′s clinical symptom did improve. His serum ACTH level came down to 40.4 pg/ml, and his selleck screening library blood pressure was controlled within 140/80 mmHg. Figure 3 Typical MRI scan changes in prolactinomas adenoma. Coronal T1-weighted postcontrast MRI scan at left and right, obtained in Patient 2, a 27-year-old woman

who presented with prolactinomas adenomas and amenorrhea-galactorrhea 4 years before undergoing MASEP GKRS. An asymmetrically enhancing mass lesion is seen in the sella turcia with extension to bilateral internal carotid artery. Patient 2′s serum prolactin level was 183.7 ng/ml. The patient was treated with MASEP GKRS twice because of the huge volume of the mass. The second MASEP GKRS was performed 1 year after the first one. The tumor was treated separately with the lower and upper part in order to protect the optic chiasma.

MRI was performed for treatment planning. 25 Gy defined to the 50% Oxymatrine isodose line is used to cover the lower part of the pituitary tumor in the first treatment, and 18 Gy defined to the 50% isodose line is used to cover the upper part of the pituitary tumor in the second time. Figure 4 Typical MRI scan changes in prolactinomas adenoma. An enhancing mass lesion is seen in the sella turcia under the T1-weighted postcontrast MRI scan performed 1 year after MASEP GKRS, but the volume of the mass had collapsed for more than 50%. Patient 2′s clinical symptom did improve. Her serum prolactin level came down to 14.5 ng/ml, and she got gestation and delivered a healthy baby recently. Figure 5 Typical MRI scan changes in GH adenoma. Coronal T1-weighted postcontrast MRI scan at upper left and right, obtained in Patient 3, a 33-year-old man who presented with GH adenomas and acromegaly 7 years before undergoing MASEP GKRS. (Figure 5) An enhancing mass lesion is seen in the sella turcia with extension into the left cavernous sinus. Patient 3′s serum growth hormone level was 497.3 ng/ml initially.

Swidsinski A, Weber J, Loening-Baucke V, Hale LP, Lochs H: Spatial organization and composition of the mucosal flora in patients with inflammatory bowel disease. J Clin Microbiol 2005, 43: 3380–3389.PubMedCrossRef 48. Bibiloni R, Mangold M, Madsen KL, Fedorak RN, Tannock GW: The bacteriology of biopsies differs between newly diagnosed, untreated, Crohn’s disease and ulcerative colitis patients. J Med Microbiol 2006, 55: 1141–1149.PubMedCrossRef 49. Lucke K, Miehlke S, Jacobs E, Schuppler M: Prevalence

of Bacteroides and Prevotella spp. in ulcerative colitis. J Med Microbiol 2006, 55: 617–624.PubMedCrossRef 50. Martinez-Medina M, Aldeguer https://www.selleckchem.com/products/ferrostatin-1-fer-1.html X, Gonzalez-Huix F, Acero D, Garcia-Gil LJ: Abnormal microbiota composition in the ileocolonic mucosa of Crohn’s disease patients as revealed by polymerase chain reaction-denaturing gradient gel electrophoresis. Inflamm Bowel Dis 2006, 12: 1136–1145.PubMedCrossRef 51. Sokol H, Lepage P, Seksik P, Doré J, Marteau P: Temperature gradient gel electrophoresis of fecal 16S rRNA reveals active Escherichia coli in the microbiota of patients with ulcerative colitis. J Clin Microbiol 2006, 44: 3172–3177.PubMedCrossRef 52. Baumgart M, Dogan B, Rishniw M, Weitzman G, Bosworth B, Yantiss R, Orsi RH, Wiedmann M,

McDonough P, Kim SG, Berg D, Schukken Y, Scherl E, Simpson KW: Culture independent analysis of ileal mucosa reveals a selective increase in invasive Meloxicam Escherichia coli of novel phylogeny relative to depletion of Clostridiales in Crohn’s disease involving the ileum. ISME J 2007, 1: 403–418.PubMedCrossRef 53. Kotlowski R, Bernstein LY2109761 concentration CN, Sepehri S, Krause DO: High prevalence of Escherichia coli belonging to the B2+D phylogenetic group in inflammatory bowel disease. Gut 2007, 56: 669–675.PubMedCrossRef 54. Andoh A, Tsujikawa T, Sasaki M, Mitsuyama K, Suzuki Y, Matsui T, Matsumoto T, Benno Y, Fujiyama Y: Fecal microbiota profile of Crohn’s disease determined by terminal restriction fragment length polymorphism analysis.

Aliment Pharmacol Ther 2009, 29: 75–82.PubMedCrossRef 55. Martinez-Medina M, Aldeguer X, Lopez-Siles M, González-Huix F, López-Oliu C, Dahbi G, Blanco JE, Blanco J, Garcia-Gil LJ, Darfeuille-Michaud A: Molecular diversity of Escherichia coli in the human gut: New ecological evidence supporting the role of adherent-invasive E. coli (AIEC) in Crohn’s disease. Inflamm Bowel Dis 2009, 15: 872–882.PubMedCrossRef 56. Dicksved J, Halfvarson J, Rosenquist M, Järnerot G, Tysk C, Apajalahti J, Engstrand L, Jansson JK: Molecular analysis of the gut microbiota of identical twins with Crohn’s disease. ISME J 2008, 2: 716–727.PubMedCrossRef 57. Ott SJ, Plamondon S, Hart A, Begun A, Rehman A, Kamm MA, Schreiber S: Dynamics of the mucosa-associated flora in ulcerative colitis patients during remission and clinical relapse. J Clin Microbiol 2008, 46: 3510–3513.PubMedCrossRef 58.

J Hosp Infect. 2014;87(2):109–14.PubMedCrossRef 18. Department

of Health. NHS reference costs 2010-2011. Department of Health 2012, United Kingdom. http://​data.​gov.​uk/​dataset/​nhs-reference-costs-2010-11 Accessed Feb 2012. 19. Welsh Healthcare Associated Infection Programme. Clostridium difficile. Mandatory Surveillance Report. Abertawe Bro Morgannwg University Health Board. Public Health Wales. 2012. http://​www2.​nphs.​wales.​nhs.​uk:​8080/​WHAIPDocs.​nsf/​3dc04669c9e1eaa8​80257062003b246b​/​87239a685280fb96​80257a4f0035fc37​/​$FILE/​ABMU%20​Report%20​Apr%20​11%20​-%20​Mar%20​12.​pdf Accessed Jul 2012. 20. Gerding DN, Muto CA, Owens RC. Measures to control and prevent Clostridium difficile this website infection. Clin Infect Dis. 2008;46:S43–9.PubMedCrossRef 21. Wiegand PN, Nathwani D, Wilcox MH, Stephens J, Shelbaya A, Haider S. Clinical and economic burden of Clostridium difficile infection in Europe: a systematic review of healthcare-facility-acquired infection. J Hosp Infect. 2012;81:1–14.PubMedCrossRef 22. Al-Eidan FA, McElnay JC, Scott MG, Kearney MP. Clostridium difficile-associated diarrhoea in hospitalised Selleckchem Seliciclib patients. J Clin Pharm Ther. 2000;25:101–9.PubMedCrossRef

23. Casari E, Ferrario A, De Luca C, Calabro M, Allibardi S, Lagioia M. Improvement in patient management through the use of a Clostridium difficile PCR real time stand alone test in acute hospital setting. In: Program and Abstracts of the 52nd International Congress of Antimicrobial Agents and Chemotherapy (ICAAC); September Tangeritin 9–12, 2012; San Francisco, CA. Abstract D-159. 24. Bartsch SM, Curry SR, Harrison LH, Lee BY. The potential economic value of screening hospital admissions for Clostridium difficile. Eur J Clin Microbiol Infect Dis. 2012;31:3163–71.PubMedCentralPubMedCrossRef 25. Brown J, Paladino JA. Impact of rapid methicillin-resistant

Staphylococcus aureus polymerase chain reaction testing on mortality and cost effectiveness in hospitalised patients with bacteraemia. Pharmacoeconomics. 2010;28:567–75.PubMed 26. Lehmann LE, Herpichboehm B, Kost GJ, Koller MH, Stueber F. Cost and mortality prediction using polymerase chain reaction pathogen detection in sepsis: evidence from three observational trials. Crit Care. 2010;14:R186.PubMedCentralPubMedCrossRef 27. Nigrovic LE, Chiang VW. Cost analysis of enteroviral polymerase chain reaction in infants with fever and cerebrospinal fluid pleocytosis. Arch Pediatr Adolesc Med. 2000;154:817–21.PubMedCrossRef 28. Eastwood K, Else P, Charlett A, Wilcox M. Comparison of nine commercially available Clostridium difficile toxin detection assays, a real-time PCR assay for C. difficile tcdB and a glutamate dehydrogenase detection assay to cytotoxin testing and cytotoxigenic culture methods. J Clin Microbiol. 2009;47:3211–7.PubMedCentralPubMedCrossRef 29. Sloan LM, Duresko BJ, Gustafson DR, Rosenblatt JE.

Adherent biofilms were stained with crystal violet, followed by ethanol solubilization of the crystal violet and quantification (A 595nm) of stained wells. The box plots (median, thick line in the box) represent the mean of 3 independent biological repeats, each assayed in quintuplicate (n = 15). *** indicates a statistically significant difference (p < 0.001), between the typA mutant and PA14 WT as determined by Whitney Mann test. However, the investigation Pirfenidone of flagellum-mediated swimming and swarming motility as well as the type IV pilus-mediated twitching motility, which are all involved in attachment and subsequent

biofilm development, revealed no differences between mutant and wild type strain (data not shown) ruling out defects in the biosynthesis and function of these cellular appendages in the typA mutant. selleck chemical Antibiotic susceptibility testing Since recent studies have demonstrated a role for TypA in multidrug resistance in E. coli[28], we studied the impact of the typA gene in antibiotic resistance of P. aeruginosa against a variety of different antimicrobial compounds. As shown in Table 1, the typA mutant exhibited a consistent 2-fold increase in susceptibility to the cationic peptides polymyxin B and colistin, the ß-lactam antibiotics ceftazidime and meropenem, as well as tetracycline in comparison to the parent

strain. This altered susceptibility could be complemented by introducing wild type copies of typA into the mutant strain. Pregnenolone No change in susceptibility was observed regarding the fluoroquinolone ciprofloxacin, the aminoglycoside tobramycin, and the cationic host defence peptide LL-37 (Table 1). Table 1 MICs of different antibiotics towards P. aeruginosa PA14 WT, PA14 typA mutant and complemented

mutant PA14 typA ::p typA +a   MIC (μg/ml) Antibiotic PA14 WT PA14typA PA14typA::ptypA + Ciprofloxacin 0.03 0.03 0.03 Meropenem 2 1 2 Ceftazidime 4 2 4 Tetracycline 8 4 8 Tobramycin 0.25 0.25 0.25 Polymyxin B 0.5 0.25 0.5 Colistin 0.25 0.125 0.25 LL-37 16 16 16 aMICs were determined by serial 2-fold dilutions in MH-medium. The MIC represents the concentration at which no growth was visually observed after 24 h of incubation at 37°C. The values shown are the modes of 4 to 6 independent experiments. Reduced virulence of PA14 typA due to down-regulation of the Type III secretion system Previous studies have shown, that uptake by and killing of eukaryotic host cells is highly dependent on the Type III secretion system in P. aeruginosa[5, 29, 30]. To analyze the potential molecular basis for reduced virulence of the typA mutant observed in our experiments, we investigated gene expression of known virulence-associated genes in P. aeruginosa using qRT-PCR on bacterial RNA of wild type and typA mutant strain isolated during host-pathogen interaction with D. discoideum.

We performed acid stress assays in the presence of these amino acids with hns-deficient strains also deleted in these genes. Only the deletion of dps led to dramatically low survival rate in the presence of arginine and lysine, while the deletion of hdeA resulted in a 5-fold decreased survival rate in the presence of arginine and slightly modified survival rate in the presence of lysine

(Table 3). Although the arginine and lysine-dependent acid resistance pathways are regulated by H-NS [1], it is not known whether AdiY and HCS assay CadC, the specific regulators of these pathways respectively, are controlled by H-NS. Real-time quantitative RT-PCR experiments were carried out on adiY and cadC with RNA isolated from FB8 wild-type and hns-deficient strains. We observed that the adiY and cadC RNA level increased five-fold in the hns mutant

(Table 4), suggesting that they may mediate the effect of H-NS on arginine and lysine-dependent acid stress resistance. To further investigate the role of adiY and cadC in the H-NS-dependent control of acid resistance, each gene was deleted in an hns background and the acid resistance assays were performed in the presence of arginine, glutamate and lysine. In the absence of adiY, much fewer bacteria survived in the presence of glutamate and arginine, but not in the presence of lysine, while Hydroxychloroquine price the cadC deletion led to extreme acid stress sensitivity only in the presence of lysine (Table 2 and 3). This suggests a role of CadC regulator in the H-NS regulation of the lysine-dependent acid stress resistance and a role of AdiY regulator in the arginine- and glutamate-dependent pathways. Table 3 Arginine and lysine-dependent acid resistance RG7420 cost of E. coli strains Strain (relevant genotype) Arginine-dependent acid resistance (% survival) Lysine-dependent acid resistance (% survival) FB8 (wild-type) 0.23 0.05 BE1411 (hns::Sm) 24.50 7.64 BE2823 (hns::Sm ΔrcsB) 4.44 1.00 BE2826 (hns::Sm Δdps) 0.11 0.28 BE2836 (hns::Sm ΔhdeA) 5.11 5.37

BE2837 (hns::Sm ΔadiY) 1.80 7.30 BE2939 (hns::Sm cadC1::Tn10) 24.24 0.001 Percentage survival is calculated as 100 × number of c.f.u. per ml remaining after 2 hours low pH treatment in the presence of arginine or lysine, divided by the initial c.f.u. per ml at time zero. Data are the mean values of two independent experiments that differed by less than 15%. Table 4 Quantitative RT-PCR analysis on H-NS targets involved in acid stress resistance   Expression ratio Gene hns/wild-type hns gadE /wild-type hns rcsB /wild-type hns hdfR /wild-type hns adiY /wild-type Glutamate-dependent specific pathway gadA 1 137.21 nd Nd 150.93 41.31 dctR 1 34.66 nd Nd 34.32 8.84 yhiM 10.75 3.41 3.40 10.90 11.36 aslB 12.92 0.66 1.10 0.69 1.32 gltD 1 1.68 nd Nd 0.48 0.52 Arginine-dependent specific pathway adiA 16.

These findings were consistent with the primary microarray-based data of this study and expression data of S. aureus Mu50 [10]. Figure 2 Transcript quantification of the essential capsule biosynthesis gene cap5E by real

time PCR. a) Transcript amounts of cap5E throughout the growth curve of hVISA SA137/93A (filled square), VISA SA137/93G (filled triangle), VSSA Newman (filled circle) and VSSA SA1450/94 (filled diamond) indicated as copy number per 106 copies of the housekeeping gene gyrB. b) Transcript amounts of cap5E of VSSA strains (R: Reynolds*, N: Newman, 14: SA1450/94) and VISA strains (A: SA137/93A*, G: SA137/93G*, Mu: Mu50) at OD600 = 1 and OD600 = 4–5 indicated as copy number per 106 copies of gyrB. * Error bars are not visible at OD600 = 1 because of minimal data variations.

The CPs of Klebsiella CP-690550 in vitro pneumoniae were found to contribute to resistance to cationic defensins, lactoferrin, protamine sulfate and polymyxin B, and in this context, the capsule was assumed to protect bacteria by limiting the interaction of the antimicrobial peptides with the surface [54]. Later, similar results were obtained with polysaccharides from Streptococcus pneumoniae GW-572016 in vivo and alginate from Pseudomonas aeruginosa [55]. A possible role of CPs in vancomycin resistance has repeatedly been discussed in the literature. Boyle-Vavra et al. found that susceptible passage revertants of the CP5 producing VISA isolates MI, NJ and PC were no longer CP typable, while passaging in presence of vancomycin retained the CP phenotype [56]. Besides, comparative expression profiling experiments on VISA isolates Mu50, MI, JH9 and their respective

susceptible parent or mutant strains showed that some (but not necessarily all) of the genes of the type 5 capsule were more highly selleck chemical expressed in the VISA strains [10, 45]. Enhanced capsule production in other VISA was also reported [57] and deletion of the yabJ-spoVG operon affected glycopeptide susceptibility and capsule production in S. aureus simultaneously [50]. Taken together, these findings encouraged us to further investigate the role of CPs in vancomycin resistance. Detection of the capsule by immunofluorescence Production of CP5 was analysed by immunofluorescent labelling of cells of SA137/93G and the susceptible strains SA1450/94 and Newman after 6 h of incubation in LB. The results revealed that the VISA strain produced higher amounts of CP5 than SA1450/94 and S. aureus Newman (Figure 3). Figure 3 Comparison of CP5 production in a VISA and two VSSA strains. CP5 was labelled by immunofluorescence (CY3, green). As a control, all cells were stained using DAPI (blue). Cells were grown for 6 h in LB at 37°C. a) VISA SA137/93G; b) control strain SA1450/94; c) S. aureus Newman.

Curr Biol 2001,11(4):258–262.PubMedCrossRef Authors’ contributions RCS, GRQS, DSN and MFN retrieved, analyzed, prepared the AtlasT4SS dataset (sequence, functional annotation, cross-references…) and illustrated the relational database. RCS and GRQS performed scripts for automated data retrieval and developed the current web pages. MFN, MOCC and CCK in cooperation carried out the CDS annotation and designed the

T4SS hierarchical classification. NCBL worked on the phylogenetic trees figures. MFN and ATRV managed the project. BMS-777607 research buy ATRV is the team leader and provides financial support. All authors read and approved the final manuscript.”
“Background Sugarcane is an efficient substrate for bioethanol production, wich is currently largely used in Brazil as a substitute for fossil fuels. Traditionally, sugarcane crops are burnt before harvest, in order to remove leaves, thus facilitating easier manual harvest. However, this procedure results in high emissions of particulate matter and smoke, which can be harmful to humans and livestock. Current Everolimus regulation of bioethanol production is leading to a transition towards mechanical harvest. Several authors have reported the positive effects of unburnt harvest (green cane) on soil fertility, soil structure, soil C levels and biological activity [1–3]. Most of these data have been generated in studies

in the Atlantic Forest biome, however none has addressed the microbial community structures and diversities in soils under burnt versus green cane management in Cerrado Biome. The Cerrado is the second largest terrestrial biome in Brazil and it is characterized by a savannah-like vegetation on ancient and plain soils [4]. Currently, cultivation of sugarcane is increasing in this region, with some states showing a 300% expansion of cropped areas over the last few years [5]. Due to high Reverse transcriptase concentrations of endemic

plant species and the accelerated pace of deforestation, the Cerrado region has been classified as a high priority area for biodiversity conservation [6]. Therefore, there is a need to develop studies that address the effects of sugarcane expansion in Cerrado soils. The use of agricultural land for cropping generally results in modifications of the soil biological and physicochemical properties, which, in turn, affect soil biogeochemical processes such as nutrient cycling and gas emissions, influencing ecosystem productivity and sustainability [7–11]. Brazil is the fifth largest contributor to the global emission of greenhouse gases (GHG). A major part, up to 75%, is the consequence of unsustainable agricultural practices next to deforestation, which include removal of crop residues, exposure of the soil surface to erosion, excessive plowing and the introduction of nitrogen fertilizers in excess [12–14].

Of the nutrient intakes that were estimated (from weighed food intake records) during the survey at baseline [5], only the major bone-related nutrient intakes (calcium, phosphorus and vitamin D) are reported here. Whereas calcium and phosphorus are derived from the diet alone, a major source of vitamin D is, of course, the action of sunlight on vitamin D precursors in the skin. About 5% of the survey participants were recorded as taking regular over-the-counter dietary supplements that contained one or more of

these three nutrients. In a previous Selleckchem BIBW2992 recent study [1], we showed that plasma zinc (amongst other redox-active nutrient status indices) robustly predicted subsequent all-cause mortality in this survey cohort. We note here that a considerable proportion selleck compound of the body’s zinc content is found in the bone, with possible implications for bone health and metabolic activity. Several recent studies have reported significant prediction of (better) survival by higher blood vitamin D status indices or vitamin D supplementation [15–26] and/or by lower serum or plasma PTH levels [15, 26–28]. Three recent

studies [7–9] have reported poorer survival at higher levels of serum calcium and/or phosphorus, usually attributed to impaired kidney function and/or inflammatory processes, and one of these [8] has also reported an association between mortality and raised serum alkaline phosphatase. Strengths and limitations of study Important strengths of the present study were that, as far as possible, the population sample was chosen as being statistically representative of the community-living people of 4-Aminobutyrate aminotransferase mainland Britain in 1994–1995. A wide range of nutrition-related factors were measured at baseline, including questionnaire-derived socio-demographic information, a 4-day weighed diet estimate, anthropometric measurements, haematology,

blood and urine biochemistry (including a large number of nutritional indices), dental assessment [29], etc.; the follow-up period for mortality outcomes was substantial, i.e. 13–14 years. On the other hand, the survey was originally designed primarily to characterise food choices and nutritional status rather than having specific focus on bone health or subsequent mortality outcomes. Another inevitable weakness, associated inevitably with any cross-sectional national survey, is the fact that the baseline measures were sampled at a single time point only. It is thus, in principle, unable to address issues of long-term causal pathways or of intervening events occurring after the baseline measures.

The number of colonies was determined using a colony counter and compared with the control (0 h) to determine bile salt tolerance. Percent survival was calculated using Equation 1. Antibacterial check details susceptibility testing Susceptibility to 24 antibiotics was determined by using the disc diffusion

method [48]. Single colonies were inoculated into M17 broth and incubated at 37°C for 24 h. A sterile cotton wool swab dipped into the bacterial suspension was used to spread bacteria evenly on the surface of M17 agar plate. Commercially available antibiotics discs (Oxide) containing penicillin G (2 units), erythromycin (10 μg), ceftriaxone (30 μg), colistin sulphate (10 μg), streptomycin (10 μg), amikacin (30 μg), norfloxacin (10 μg), chloramphenicol (30 μg), tetracycline (10 μg), nalidixic acid (30 μg), ampicillin (25 μg), gentamycin (30 μg),

mecillinam (25 μg), nitrofurantoin (300 μg), sulfamethoxazole/trimethoprim (25 μg), vancomycin (30 μg), kanamycin (30 μg), neomycin (30 μg), lincomycin (10 μg), cloxacillin (5 μg), ciprofloxacin (10 μg), cefuroxime sodium (30 μg), bacitracin (10 μg), or novobiocin (30 μg) were carefully placed on the surface Metformin concentration of the dried agar plates to ensure uniform contact between the disc and agar. The plates were then incubated at 30°C for 24 h. Inhibition zones (including the disc diameter) were measured, and isolates were categorized as sensitive (≥ 21 mm), intermediate (16–20 mm), or resistant (≤ 15 mm), as previously described [29, 49]. β-galactosidase activity The method described by Karasova et al.[50] was used to

test for β-galactosidase activity. The isolate was incubated at 37°C for 24 h on an MRS agar plate containing 0.01% X-gal (5-bromo-4-chloro-3-indolyl β-D-galactopyranoside, Vivantis, Malaysia) and 0.1 mM IPTG (isopropyl β-D-1-thiogalactopyranoside, Vivantis) dissolved in dimethyl sulfoxide. Identification of isolates using API 50 CHL API 50 CHL strips (API systems, bioMérieux, France) were used to characterize the isolates, according to the manufacturer’s Carnitine dehydrogenase instructions. The inoculated strips were incubated at 30°C, and the reactions were observed after 48 h. The API database (bioMérieux SA) and accompanying computer software were used to interpret the results. Readings were taken after a 48-h incubation at 30°C. Growth on a particular substrate changed the color of the medium from violet to yellow, which was scored on a 5-point scale (intense yellow = 5). A score ≥3 was considered a positive result. The test was performed in triplicate. Identification of isolates by 16S rDNA sequencing and phylogenetic analysis The isolates were identified by 16S rDNA sequencing to confirm the results obtained from biochemical identification. Briefly, the procedure is as follows. DNA extraction DNA was extracted using the method described by Leenhouts et al.[51], with some modifications. Cells harvested from an overnight culture (1.