text:
Serviços Personalizados
Links relacionados
- Citado por Google
- Similares em Google
Water SA
versão On-line ISSN 1816-7950
versão impressa ISSN 0378-4738
Water SA vol.35 no.1 Pretoria Jan. 2009
Suitability of total coliform β-D-galactosidase activity and CFU counts in monitoring faecal contamination of environmental water samples
VC WutorI, *; CA TogoII; BI PletschkeII
IAgriculture and Agri-Food Canada, Lethbridge Research Center, PO Box 3000, Lethbridge, Alberta, Canada T1J 4B1
IIDepartment of Biochemistry, Microbiology and Biotechnology, Rhodes University, PO Box 94, Grahamstown 6140, South Africa
Total coliforms are a group of bacteria found in high numbers in mammalian intestines; hence their presence in water indicates the possible contamination with faecal material. Total and faecal coliform counts were monitored over a period of 18 months using mFC, m-Endo and CM1046 media together with enzymatic assays on 215 environmental water samples obtained from the Eastern Cape Province of South Africa. A positive correlation, with an R2 value of 0.9393 was observed between faecal and total coliform colony units employing mFc and m-Endo media, and 0.8818 using CM1046 media. Also, a positive correlation was observed between Escherichia coli colony-forming units and β-d-galactosidase (B-GAL) activity (R2=0.8542). Overall, this study indicated that faecal contamination of environmental water samples could be monitored by measuring total coliform β-galactosidase activity and total coliform colony-forming units.
Keywords: colony-forming units (CFUs), coliforms, E. coli, β-d-galactosidase (B-GAL), β-D-glucuronidase (GUD)
Coliforms are a group of bacteria employed to assess the microbiological quality of water and/or food. Most faecal coliforms are present in large numbers among the intestinal flora of human and other warm-blooded animals, and are thus commonly found in faecal material. As a result, the presence of these non-pathogenic microorganisms in water indicates the potential presence of pathogenic microorganisms of faecal origin (Venter, 2000; Rompré et al., 2002) and consequently a health hazard. It is estimated that 80% of communicable diseases in the world are waterborne (Devi et al., 2008). Analysing for total coliform bacteria and Escherichia coli is the most common method used to test the hygienic quality of drinking water (Huertas et al., 2003; Pitkänen et al., 2007). Bacteria as diverse as Escherichia coli, Citrobacter, Klebsiella, Enterobacter, Serratia, Erwinia and Yersinia are all positive for the traditional coliform test (Standard Methods, 2005). Among the afore-mentioned bacteria, only Escherichia coli appear to be of faecal origin (Gofti et al., 1999; Rompré et al., 2002; Schraft and Watterworth, 2005) (Fig. 1).
However, there have been criticisms on the suitability of total coliforms, as markers of faecal contamination of water and/or food (Gofti et al., 1999; Leclerc et al., 2001; Stevens et al., 2001; Rose and Grimes, 2002; Hörman and Hänninen, 2006). Studies on total coliforms reveal that they can have an environmental origin besides faecal sources, thus these organisms may not be suitable indicators of faecal contamination (Leclerc et al., 2001). Escherichia coli and enterococci have been shown to be consistently associated with faecal pollution and thus the most reliable indicators of faecal contamination (Leclerc et al., 1999; 2001; Gofti et al., 1999; Rompré et al., 2002; Schraft and Watterworth, 2005). Furthermore, there have been occurrences of total coliforms in the absence of documented waterborne morbidity in the community, and outbreaks where coliforms have not been found (McFeters and Camper, 1983; McFeters et al., 1986). Although the latter could be due to the fact that the currently accepted methods may lead to underestimations of waterborne coliforms (Hudson et al., 1983; McFeters et al., 1986), this does not rule out the unreliability of coliforms as faecal indicators. Despite these setbacks, total coliforms are still being used in national and international guidelines for the microbiological quality assessment of water (Grabow et al., 1999; Noble et al., 2003; Horman et al., 2004; Byamukama et al., 2005).
The presence of total coliforms and E. coli (faecal coliform) could be detected by measuring the activity of β-d-galactosidase (B-GAL) and β-D-glucuronidase (GUD) enzymes respectively (Wutor et al., 2007a). These enzymes hydrolyse a wide range of chromogenic and fluorogenic substrates yielding products which can be monitored spectrophotometrically and fluorometrically, respectively. In this study, the chromogenic substrates chlorophenol red β-D-galactopyranoside (CPRG), and p-nitrophenyl-β-D-glucuronide (PNPG) were employed to measure B-GAL and GUD activities respectively, while mFc, m-Endo and CM 1046 media were used in CFU counts.
In this paper, we examine the relationship between faecal coliform (FC) and total coliform (TC), thus the potential of using TC β-d-galactosidase activity and CFU counts as a measure of faecal coliform contamination of environmental water samples.
Materials and methods
Chlorophenol red β-D-galactopyranoside (CPRG), disodium hydrogen phosphate, dihydrogen sodium phosphate were all obtained from Merck (Darmstadt, Germany), p-nitrophenyl-β-D-glucuronide (PNPG) (Merck, Darmstadt, Germany). The media, mFc, mEndo and CM1046 were obtained from Merck (Darmstadt, Germany), Fisher Scientific (Loughborough, UK) and Oxoid (Hampshire, UK) respectively. The buffers were prepared using water purified with a Milli-Q system (Millipore, Milford, CT, USA). All reagents were of the highest analytical grade available.
Water collection was performed in accordance with the standard procedures outlined in Standard Methods, 2005. Water samples were collected weekly from two points along the Bloukrans River, Grahamstown in the Eastern Cape of South Africa as well as from other sources in the Eastern Cape Province of South Africa for a period spanning 18 months between 2005 and 2006. Samples were collected aseptically in 1 ℓ sterile Pyrex bottles (Schott Duran, Germany), placed on ice and transported immediately to the laboratory and analysed within an hour. Samples were collected in triplicates between 08:00 and 08:30 on each sampling day. In total, 215 samples were studied.
Enzyme Assays
The B-GAL assay was performed as described in Wutor et al. (2007b). Briefly, CPRG (80 µg in 20 µℓ water) was mixed with 90 µℓ 0.1 M sodium phosphate buffer (pH 7.8) in a 96-well, flat-bottomed micro-titre plate, and 90 µℓ of environmental (water) sample added to initiate the reaction. The change in absorbance was monitored at 575 nm over 24 h on a PowerWavex (Bio-Tek Instruments, USA).
Togo et al. (2006) described the GUD assay employed in the study. PNPG (10 mM) was prepared in 0.1 M Tris-HCl, pH 8.0 containing 0.6 mM CaCl2 buffer. Assay buffer (50 µℓ) and 90 µℓ enzyme solution/water sample were mixed in a 96-well, flat-bottomed micro-titre plate. The reaction was initiated by the addition of 110 µℓ PNPG. Kinetic readings (24 h at 1 h intervals) were performed at 405 nm and 25 ºC using a Power Wavex spectrophotometer (Bio-Tek Instruments, USA).
In each enzyme assay, 2 sets of controls were set up; an enzyme control and a substrate control. All readings were taken against milli-Q water (Millipore Corporation, USA). All assays were performed in triplicate, unless otherwise stated. All results are reported as means ± standard deviations.
Microbiological analyses
Water samples of 100 mℓ volumes were filtered through membrane filters (Whatman 0.45 µm pore size) and placed on mFc plates for the detection of faecal coliforms at 44.5°C over 24 h; CM 1046 media (selective E. coli/coliform chromogenic medium) and m-Endo-agar plates (total coliforms) and incubated at 37°C for 24 h, after which colonies were counted. All colonies which developed a pink colour were counted as other coliforms while those that developed a purple colour were counted as E. coli on the CM 1046 media (the sum of both pink and purple colony counts gave the total coliform load of the water sample under consideration) while red colonies with metallic reflections were counted as total coliforms (m-Endo agar) and colonies with light to dark blue colonies were counted as faecal coliforms (mFc agar). These were then expressed as coliform colony-forming units (CFU) per 100 mℓ (Wutor et al., 2007a).
Results and discussion
The source of B-GAL enzyme was validated as from E. coli by using SDS-PAGE and MALDI-ToF (data not shown). The MALDI-ToF technique has been used successfully in the analysis of proteins, peptides, glycoproteins, oligosaccharides, and oligonucleotides and has the capability of measuring masses to within 0.01% of the molecular mass of the sample (Takach et al., 1997; Whitelegge, 2003; Kammerera et al., 2005). Enzymatic assays were performed alongside growth on selected media to correlate enzymatic activity and colony counts.
Wutor et al. (2007a) in a previous study observed a positive correlation between total coliform CFUs and B-GAL activity; R2 of 0.941 for CM 1046 media, and 0.7051 for m-Endo media, while Togo et al. (2006) established a correlation of 0.8909 between GUD activity and E. coli CFUs.
Detection of TC and FC using media
Generally, we observed a significant (P<0.05) correlation between TC and FC on m-Endo and mFc media with a R2 value of 0.9392 and a R2 value of 0.8818 for TC and FC on CM 1046 media (Figs. 2 and 3). Though different media were employed in this analysis, the benefits of an early warning signal on the microbiological status of water from assessing only total coliforms cannot be underestimated.
Detection of TC and FC
A positive correlation, with an R2 value of 0.8542 at 95% confidence interval was observed for E. coli colony counts and B-GAL activity (Fig. 4). It must be noted that this value, though lower than the previous statistically, yet still high, could be accounted for by the fact that not all E. coli cells will grow on the media, a major limitation in the use of the conventional plating procedure. However, those viable but non-culturable (VBNC) bacteria will still produce the B-GAL enzyme, thus making the enzyme assays better indicators than the plating on media (Pletschke et al., 2006; 2008). One major limitation in product development is the transferring of research from the laboratory to the field. In this work it is important to note that results were obtained from directly using the environmental samples to try easing the transfer.
Farnleitner et al. (2001) and Garcia-Armisen et al. (2005) reported R2 values of 0.73 and 0.92 respectively, in correlation studies between GUD and E. coli. Variations in correlation coefficients for same enzyme can be attributed to differences in climatic conditions and the environments from which the samples were collected. Both factors dictate the nature and composition of interfering compounds in water samples. Furthermore, enzyme substrates are susceptible to interference from pollutants during analysis. For example, Farnleitner et al. (2002) employed 4-methylumbelliferyl-β-D-glucuronide, a fluorogenic substrate in their study, while the same substrate yielded unreliable results in our study (results not shown). Therefore, it is predicted that the future of water quality monitoring is likely to rely on a matrix approach placing less emphasis on any one single parameter (Ashbolt et al., 2001; Havelaar et al., 2001; Niemela et al., 2003; Van Heerden et al., 2005). Thus, the utilisation of the GUD and B-GAL assays remains relevant as complementary or presumptive tests for faecal pollution.
In conclusion, one could use total coliform β-D-galactosidase enzyme activity and CFU determination as a very good measure of microbial water quality assessment. Total coliform measurement may be considered as an imperfect but useful criterion of drinking water quality assessment and should find use at both small and large treatment facilities, as well as in routine analysis.
The authors would like to thank the Water Research Commission (WRC) of South Africa for financial assistance during the course of this study.
ASHBOLT NJ, GRABOW WOK and SNOZZI M (2001) Indicators of microbial water quality. In: Fewtrell L and Bartram J (eds.) Water Quality: Guidelines, Standards and Health. Risk Assessment and Management for Water-Related Infectious Disease. IWA Press, London, UK. 289-316. [ Links ]
BYAMUKAMA D, MACH RL, KANSIIME F, MANAFI M and FARNLEITNER AH (2005) Discrimination efficacy of fecal pollution detection in different aquatic habitats of a high-altitude tropical country, using presumptive coliforms, Escherichia coli, and Clostridium perfringens spores. Appl. Environ. Microbiol. 71 65-71. [ Links ]
DEVI R, ALEMAYEHU E, SINGH V, KUMAR A, and MENGISTIE E (2008) Removal of fluoride, arsenic and coliform bacteria by modified homemade filter media from drinking water. Bioresour. Technol. 99 2269-2274. [ Links ]
FARNLEITNER AH, HOCKE L, BEIML C, KAVKA GG and MACH RL (2002) Hydrolysis of 4-methylumbelliferyl-β-d-glucuronide in differing sample fractions of river waters and its implication for the detection of faecal pollution. Water Res. 36 975-981. [ Links ]
FARNLEITNER AH, HOCKE L, BEIWL C, KAVKA GG, ZEICHMEISTER T, KIRSCHNER AKT and MACH RL (2001) Rapid detection of Escherichia coli contamination in polluted water. Lett. Appl. Microbiol. 33 246-250. [ Links ]
GARCIA-ARMISEN T, LEBARON P and SERVAIS P (2005) β-D-glucuronidase activity assay to assess viable Escherichia coli abundance in fresh waters. Lett. Appl. Microbiol. 40 278-282. [ Links ]
GOFTI L, ZMIROU D, SEIGLE MF, HARTEMANN P and POTELON JL (1999) Waterborne microbiological risk assessment: a state of the art and perspectives. Epidemiol. Rev. 47 (1) 61-73. [ Links ]
GRABOW WOK, BOTMA KL, DE VILLIERS JC, CLAY CG and ERASMUS B (1999) Assessment of cell culture and polymerase chain reaction procedures for the detection of polioviruses in wastewater. Bull. World Health Organ. 77 973-980. [ Links ]
HAVELAAR A, BLUMENTHAL U, STRAUSS M, KAY D and BARTRAM J (2001) Guidelines: the current position. In: Fewtrell L and Bartram J (eds.) Water Quality: Guidelines, Standards and Health. World Health Organization and IWA Publishing, London, UK. [ Links ]
HÖRMAN A and HÄNNINEN M (2006) Evaluation of the lactose Tergitol-7, m-Endo LES, Colilert 18, Readycult Coliforms 100, Water-Check-100, 3M Petrifilm EC and DryCult Coliform test methods for detection of total coliforms and Escherichia coli in water samples. Water Res. 40 3249-3256. [ Links ]
HÖRMAN A, RIMHANEN-FINNE R, MAUNULA L, VON BONSDORFF CH, TORVELA N, HEIKINHEIMO A and HÄNNINEN ML (2004) Campylobacter spp., Giardia spp., Cryptosporidium spp., Noroviruses and indicator organisms in surface water in Southwestern Finland, 2000-2001. Appl. Environ. Microbiol. 70 (1) 87-95. [ Links ]
HUDSON LO, HARKLIN JW and BATTAGLIA M (1983) Coliforms in water distribution system a remedial approach. J. Am. Water Works Assoc. 75 564-568. [ Links ]
HUERTAS A, BARBEAU B, DESJARDINS C, GALARZA A, FIGUEROA MA and TORANZOS GA (2003) Evaluation of Bacillus subtilis and coliphage MS2 as indicators of advanced water treatment efficiency. Water Sci. Technol. 47 255-259. [ Links ]
KAMMERERA B, FRICKENSCHMIDT A, GLEITER CHM, LAUFER S and LIEBICH H (2005) MALDI-TOF MS Analysis of Urinary Nucleosides. J. Am. Soc. Mass Spectr. 16 (6) 940-947. [ Links ]
LECLERC H, DEVRIESE LA and MOSSEL DAA (1999) Taxonomical changes in intestinal (faecal) enterococci and streptococci: consequences on their use as indicators of faecal contamination in drinking water. J. Appl. Bacteriol. 81 459-466. [ Links ]
LECLERC H, MOSSEL DAA, EDBERG SC and STRUIJK CB (2001) Advances in the bacteriology of the coliform group: Their suitability as markers of microbial water safety. Ann. Rev. Microbiol. 55 201-234. [ Links ]
McFETERS GA and CAMPER AK (1983) Enumeration of indicator bacteria exposed to chlorine. Adv. Appl. Microbiol. 29 177-193. [ Links ]
McFETERS GA, KIPPIN JS and LECHEVALLIER MW (1986) Injured coliforms in drinking water. Appl. Environ. Microbiol. 51 (1) 1-5. [ Links ]
NIEMELA SI, LEE JV and FRICKER CR (2003) A comparison of the International Standards Organisation reference method for the detection of coliforms and Escherichia coli in water with a defined substrate procedure. J. Appl. Microbiol. 95 (6) 1285-1292. [ Links ]
NOBLE RT, MOORE DF, LEECASTER MK, McGEE CD and WEISBERG SB (2003) Comparison of total coliform, faecal coliform, and enterococcus bacterial indicator response for ocean recreational water quality testing. Water Res. 37 1637-1643. [ Links ]
PITKÄNEN T, PAAKKARI P, MIETTINEN IT, HEINONEN-TANSKI H, PAULIN L and HÄNNINEN M (2007) Comparison of media for enumeration of coliform bacteria and Escherichia coli in non-disinfected water. J. Microbiol. Methods. DOI: 10.1016/j.mimet.2006.10.007. [ Links ]
PLETSCHKE B, TOGO C and WUTOR V (2006) On-Line Real-Time Enzyme Diagnostic System for the Detection and Monitoring of Faecal Contamination of Water Intended for Drinking Purposes. WRC Report No. 1446/1/06. Water Research Commission, Pretoria, South Africa. [ Links ]
PLETSCHKE B, TOGO C, WUTOR V and LIMSON J (2008) On-Line Real-Time Enzymatic Biosensor System for the Rapid Detection of Faecal Contamination of Water Intended for Drinking Purposes. WRC Report No. 1603/1/08. Water Research Commission, Pretoria, South Africa. [ Links ]
ROMPRÉ A, SERVAIS P, BAUDART J, DE-RAUBIN MR and LAURENT P (2002) Detection and enumeration of coliforms in drinking water: current methods and emerging approaches. J. Microbiol. Methods 49 31-54. [ Links ]
ROSE JB and GRIMES DJ (2002) Reevaluation of Microbial Water Quality: Powerful New Tools for Detection and Risk Assessment. American Society of Microbiology, Washington, DC. [ Links ]
SCHRAFT H and WATTERWORTH LA (2005) Enumeration of heterotrophs, fecal coliforms and Escherichia coli in water: comparison of 3M™ Petrifilm™ plates with standard plating procedures. J. Microbiol. Methods 60 (3) 335-342. [ Links ]
STANDARD METHODS (2005) Standard Methods for the Examination of Water and Wastewater (20th edn.) Clesceri LS, Greenberg AE and Eaton AD (eds.). American Public Health Association, Washington DC. [ Links ]
STEVENS M, ASHBOLT N and CUNLIFFE D (2001) Microbial Indicators of Water Quality: An NHMRC Discussion Paper. National Health and Medical Council Act, Canberra. [ Links ]
TAKACH EJ, HINES WM, PATTERSON DH, JUHASZ P, FALICK AM, VESTAL ML and MARTIN SA (1997) Accurate Mass Measurements Using MALDI-TOF with Delayed Extraction. Communications from the 11th Int. Conf. on Methods in Protein Structure Analysis. J. Prot. Chem. 16 (5) 363-369(7). [ Links ]
TOGO C A, WUTOR VC and PLETSCHKE BI (2006) Properties of in situ Escherichia coli β-D-glucuronidase (GUS): evaluation of chemical interference on the direct enzyme assay for faecal pollution detection in water. Afr. J. Biotechnol. 5 (22) 2338-2344. [ Links ]
VAN HEERDEN J, EHLERS MM, HEIM A and GRABOW WOK (2005) Prevalence, quantification and typing of adenoviruses detected in river and treated drinking water in South Africa. J. Appl. Microbiol. 99 (2) 234-242. [ Links ]
VENTER SN (2000) Rapid Microbiological Methods: The Status Quo. Blue Pages, IWA, London, UK. [ Links ]
WHITELEGGE JP (2003) Thylakoid membrane proteomics. Photosynth. Res. 78 (3) 265-277. [ Links ]
WUTOR VC, TOGO CA and PLETSCHKE BI (2007A) Comparison of the direct enzyme assay method with the membrane filtration technique in the quantification and monitoring of microbial indicator organisms seasonal variations in the activities of coliforms and E. coli, temperature and pH. Water SA 33 (1) 107-110. http://www.wrc.org.za/downloads/watersa/2007/Jan%2007/2049.pdf [ Links ]
WUTOR VC, TOGO CA and PLETSCHKE BI (2007B) The effect of physico-chemical parameters and chemical compounds on the activity of β-d-galactosidase (B-GAL), a marker enzyme for indicator microorganisms in water. Chemos. 68 622-627. [ Links ]
Received 18 August 2008; accepted in revised form 8 January 2009.
* To whom all correspondence should be addressed
+1 (403) 3173358; fax: +1 (403) 3823156;
e-mail: email@example.com


text:
New Sport models join lineup of sporty sedans.
Midsize Cars
Editor: Sam Moses
$18,999 Base Price MSRP
$26,199 As Tested MSRP
Model Lineup: "Suzuki Kizashi S ($18,999); SE ($22,049); GTS ($23,049); SLS ($24,849)"
Standard Engine: 2.4-liter inline-4
Engine Options: 2.4-liter I4
Standard Transmission: 6-speed manual
Transmission Options: 6-speed manual; 6-step CVT
Basic Warranty: "3 years/36,000 miles"
Assembled In: Japan
Model Tested MSRP: "Suzuki Kizashi SLS ($24,849)"
Standard Equipment: "dual-zone auto climate control with rear seat vents, leather upholstery with contrasting stitching, sport seats, heated power seats, power windows and door locks, remote entry, pushbutton start, 10-speaker Rockford Fosgate audio with steering wheel controls, ambient and footwell lighting, rear seat pass-through to trunk, aero body kit including front fascia, side sills and spoiler, 18-inch alloy wheels with wide profile tires, sport tuned suspension, three-spoke leather-wrapped steering wheel; rear parking sensor, HomeLink, auto-dimming rearview mirror, heated mirrors, auto headlights"
Options As Tested: "premium floor mats ($125), metallic paint ($130), XM radio ($350)"
Standard Safety Equipment: "dual-stage frontal airbags, side-impact airbags front and rear, airbag curtains, advanced electronic stability control with ABS, tire pressure monitor"
Optional Safety Equipment: all-wheel drive; rearview cameraDestination Charge: $745
Stacking its quality up against a full field of four-cylinder sports sedans, Suzuki last year put the performance brands of the world on notice that they have some unexpected company. Suzuki wasn't just bragging, as the Kizashi has been a resounding sales and driving success.
Kizashi uses one engine, a good 2.4-liter aluminum four-cylinder with variable valve timing that does the job well, getting about 25 combined mpg, by both EPA ratings and our couple weeks in Kizashi models. It runs smooth and makes 185 or 180 horsepower, depending on whether it's mated to the 6-speed manual transmission or Continuously Variable Transaxle, which operates automatically or can be shifted through six steps using paddles on the steering wheel.
Suzuki engineers invested much time and attention in the development of the suspension, MacPherson strut front and multi-link rear, and it shows; the ride, including over rough roads, and handling, including hard driving on the track, are exceptional for a midsize sports sedan that costs a mere $20,000. The chassis raises the bar for rigidity. The brakes, too, we found to be pitch perfect.
All-wheel drive is an option. It's a sophisticated system that turns on and off, like some SUVs, so that you can use the normal front-wheel-drive on dry pavement and not waste fuel. Called i-AWD, the all-wheel-drive package, available with the CVT, brings Suzuki to the game with the big boys, such as the Subaru Legacy and Audi A3.
Styling-wise, the Kizashi looks both potent and elegant, in its modest midsize manner. Lovely lines: a touch of Audi here, a pinch of Lexus there. No overachieving swoops or flares, no gratuitous chrome trying to grab your attention.
New for 2011 are the Kizashi GTS and Kizashi SLS models. Both feature special Sport bodywork, including a more muscular front fascia and lower grille, side sill extensions, spoiler on the trunk lid, and 18-inch alloy wheels. The crossed mesh grille is titanium colored.
Inside the cabin, the quality of the materials is high, maybe especially the standard sport seats. The layout of the controls and instruments is clean, and the standard equipment is plentiful, including climate control vents for the rear seat. Even the base model has remote entry, pushbutton start, power windows, USB port, and other equipment many base models lack, although cruise control is not included at that lowest price. Both Sport models get their own steering wheels, and the SLS Sport gets contrasting stitching on its leather seats.
Kizashi straddles the line between compact and midsize sedans. Popular Mechanics magazine included it in a face-off of midsize sedans in 2010, and the Kizashi scored first-place in handling, braking, skid pad grip, and acceleration, against Honda Accord, Toyota Camry, Ford Fusion, Subaru Legacy, Nissan Altima, Mazda 6, and Chevrolet Malibu.
Safety-wise, the Kizashi leads the field, with no less than eight standard airbags, advanced stability control using the ABS, projector beam headlamps, and a tire pressure monitor. The Kizashi has surpassed the government's 2014 standards for side pole impact and offset front collision."
"The 2011 Suzuki Kizashi comes in four models, Kizashi S, SE, GTS Sport, and SLS Sport, all of which use the same 2.4-liter engine.
Kizashi S ($18,999) comes with auto dual-zone climate control with rear seat vents, cloth upholstery, sport seats, fold-down pass-through rear seat, nine-speaker AM/FM/CD with MP3, steering-wheel audio controls, USB port, projector beam headlamps, keyless pushbutton start, ambient footwell lighting, power locks, power windows, power mirrors, illuminated visor mirror, intermittent wipers, security system, 16-inch steel wheels. Kizashi S is available with a 6-speed manual or CVT ($21,049); the latter includes cruise control and leather-wrapped steering wheel. Kizashi S is also available with all-wheel drive and the CVT ($22,099).
Kizashi SE ($22,049) comes only with the CVT and adds 10-way power driver's seat with three memory positions, leather-wrapped steering wheel, shift knob, and parking brake lever, paddle shifters, cruise control, wide-profile 18-inch tires on alloy wheels. Kizashi SE is available with AWD ($23,399).
Kizashi Sport GTS ($23,049) features sporty body work, including a muscular front fascia and lower grille, body side sill extensions, trunk spoiler, steering wheel, sport bumpers, fancy 18-inch alloy wheels. The GTS also adds a moonroof and fog lights, 10 speakers, 425-watt Rockford Fosgate system and iPod connectivity. The GTS 6-speed manual gets a higher numerical 4.11 final drive ratio (quicker acceleration performance); also available is the CVT ($24,149) or CVT AWD ($25,499).
Kizashi Sport SLS ($24,849) upgrades with leather upholstery with contrasting stitching, heated seats, power passenger seat, heated mirrors, auto headlights, auto-dimming rearview mirror, rear parking sensor, HomeLink garage door opener, rain-sensing wipers. The SLS offers a choice of 6-speed manual, CVT ($25,949), or CVT AWD ($27,299). XM Satellite Radio ($350) is available.
Safety equipment includes eight airbags: two frontal, two side curtain, and one for each door. Active safety features include antilock brakes (ABS), electronic stability control, electronic brake-force distribution. The government-mandated tire monitor is there, also. Suzuki says the Kizashi already meets some of the 2014 crash standards, including side pole crash and front offset collision. More than 100 Kizashis were crashed by Suzuki during testing. Optional all-wheel drive can enhance safety in slippery conditions. The basic warranty is 3 years or 36,000 miles, but the powertrain is covered for 7 years or 100,000 miles."
Walk Around
"The styling of the Kizashi is Suzuki all the way, especially head-on. Kizashi's looks are consistent with its performance profile. Not a would-be or wannabe sports sedan, but a modest sports sedan on the way up. Nothing is overdone. Well, maybe just a little bit, the twin chrome integrated angular tailpipes. A touch of cool. We're okay with that. The 18-inch alloy wheels are eye-catching.
The Kizashi has a rounded, Audi-like stance. In profile, especially the roofline and front valance with the S and SE models, it's Lexus-like, with a neat little nose. The hood is a small bulge, with no extra contours other than its natural overall shape. The shape of the grille is totally Suzuki, as if that big S in the center didn't already say so. Headlights fit just right, and the lines are clean to the three other air openings, with the two outside slots holding the fog lights.
All SLS and GTS models come with Sport trim, which includes special bodywork. The new fascia on the Sport models is full and rounded, reminiscent of the aerodynamic nose on the Kizashi that Suzuki and Road & Track magazine took to Bonneville and set a class record with, an astounding 203.7 mph, thanks mostly to a big supercharger.
It's quite tidy from the rear, although you can't really tell the Kizashi is special until you see the chrome Kizashi nameplate, in racy Japanese-y script."
"Kizashi's interior is comfortable and free of small inconveniences, and that's saying a lot. High-quality materials make even the base Kizashi S feel like a more expensive car. The layout of the center stack is slick and efficient, finished in black and aluminum plastic. You can easily understand and operate the knobs and controls, everything in its logical place, handsomely so.
The instrumentation isn't as clean as it might be, with dozens of little lines as increments on the tachometer and speedo, running all the way to a dreamy 150 mph; and there's a reflection at night that blurs the edges of the smaller gauges. There's a 2x4-inch digital display between the tach and speedo, with access to everything from outside temp to instantaneous fuel mileage. We mostly watched our average mpg, which ended up 23.8 mpg.
The Sport SLS that we drove had handsome cross-stitched leather seats with high-density, low-fatigue foam. The 425-watt 10-speaker Rockford Fosgate sound system sounded beautiful, but the FM reception was often weak (though this was in bad weather). The leather-wrapped shift lever feels good.
If your Kizashi has the CVT, you'll be using the paddle shifters a lot. The paddles are sized and shaped just right, unlike some. There's a nice 3-spoke steering wheel with controls; it feels good in your hands when driving through the twists. Fold-down rear headrests improve rearview visibility. Overhead and footwell ambient lighting is cool.
We've put a lot of miles on various Kizashi models. There was one all-day run on choppy pavement in the Sport models with lots of turns to toss your butt and shoulders around, and another couple hundred miles on a dark curving interstate in a hard long drizzle, with puddles all over the freeway. The fit of the seat made it all no sweat, and we enjoyed the hours and concentrated driving.
We got more seat time in an S model with cloth upholstery that was smooth and elegant in charcoal. If you skip the leather you'll still have a stylish car. You can keep the Kizashi's price near base MSRP, and still get the good stuff.
There are cupholders in front, and in the rear they slide forward out of the dropped center armrest. Nice leather door grab handles. A standard folding rear seat that opens up the trunk is always useful in a car; in addition, there's a hatch to the trunk for skis, 2x4s, and other long cargo.
The legroom in the rear is adequate, although it's a couple inches less than the redesigned Subaru Legacy, a competitor with a base model at that same under-20K price (and including all-wheel drive). The Legacy is also a couple inches longer in wheelbase and length. But what the Kizashi lacks in legroom, it might make up in temperature comfort, with that rear seat climate vent.
There's a lot of sound insulation, so the cabin is very quiet. The Kizashi offers extensive corrosion protection, including resin panels under the body, zinc-plated steel sheets, and hot wax imbedded in the suspension mounting points.
Kizashi comes with keyless ignition: Press the Start button to fire it up. We prefer a traditional key."
Driving Impressions
"Chassis and suspension are the best parts of the driving impressions. And brakes. Engine is good too. That about covers everything.
The Kizashi was developed on the Autobahn, alpine roads in Switzerland, cobblestone streets in England, and on the Nurburgring racing circuit. The firm KYB shock absorbers don't swallow the bumps or smooth them out, but neither do they transmit them. They follow the undulations in the road, and that's not uncomfortable but confidence inspiring. You know they're paying attention. At high speeds on a curvy freeway, you can still feel the shocks working. The car moves a bit as it follows the road, so you have to stay alert.
Suzuki put a great deal of time and pride into the suspension and chassis with torsional rigidity higher than some European competitors. The Kizashi chief engineer, Hide Kumashiro, a former motorcycle road racer, stressed handling as his highest priority, which is why high-performance KYB rear shocks are used, with a carefully designed multi-link rear suspension with imbedded aluminum. We pushed the Kizashi on the road, over undulating and sometimes rough surfaces, quick changes of direction under braking, and it never gave us an unsatisfying moment of wobble or softness. Nor did it ever jar us, not once, which might be saying even more. We weren't in the Alps, merely the Gifford Pinchot National Forest around Washington's Mount Adams, but we'll take it.
The Kizashi feels sportier than the Mazda6 we recently tested, especially the engine. It's smooth, sharp, and sounds nice, from zero to 6500 rpm redline, but 6000 rpm is a sweet spot to upshift with the 6-speed gearbox. The engine characteristics are steady. Against its competitors, the 185-horsepower Kizashi is solid; by comparison, the Acura TSX has 201 hp, the Mazda6 170 hp. But the Kizashi is quicker from zero to 60 than either of them, and a lot cheaper than the TSX. It also comes with way more standard equipment.
We also drove a Kizashi with the CVT, running it hard for about 80 challenging, curvy miles. The CVT sucks 5 horsepower and brings the redline down to 6000 rpm, and compromises the sportiness. It shifts with paddles through six steps (like gears in a gearbox, but not), and you have to shift like mad to keep the engine in the powerband, even with good torque.
At full throttle, the engine jumps to 6000 rpm and just stays there and buzzes away as the car gains speed. The response of the CVT is sharp, but it just changes the power delivery too much, if what you want is the throttle feel of an old-school sports sedan. It changes the whole dynamic and sound of the engine. But if you don't care about driving hard over curving roads, you'll be happier with the CVT. Around town, you can forget it, or you can use the paddles if you want. In that way it's like an automatic transmission, only more efficient.
We found the SLS with the 6-speed manual quiet under acceleration and even quieter at a cruising speed. Eighty miles per hour is only 3000 rpm in 6th gear, and the car might as well be gliding, for all its smoothness. And because the torque is a healthy 170 pound-feet at 4000 rpm, there's enough torque at lower rpm in 6th gear to accelerate when needed, without downshifting all the time. While we were at it, we got nearly 25 mpg cruising at 80 mpg.
We got about a dozen hot laps around Portland International Raceway in the Kizashi. A rev limiter intrudes (mildly) at 6500 rpm, so shifting at 6000 rpm works well. The clutch and linkage are neither short-throw nor aggressive; in fact the clutch feels a bit soft, but that's not a bad thing. It's a mild sports sedan.
On Portland International Raceway, it didn't understeer. That's rare for any front-wheel-drive car, including some of the expensive sports sedans. Suzuki brought a test mule to the launch, a Kizashi fitted with a Suzuki/GM 3.6-liter V6 engine making more than 250 horsepower, and we took some laps in that hotrod; driven with appropriate restraint, it didn't understeer either. No V6 is planned for the Kizashi, but the point was proven, that the Kizashi suspension is built to take a lot more horsepower. A turbocharged Kizashi is said to be on the way.
Fuel economy is an EPA-estimated 23/30 mpg City/Highway for a Suzuki Kizashi SE with the CVT automatic. The CVTs generally get better fuel economy than the manual transmissions, beating them by 2-3 mpg around town. All-wheel drive costs about one mpg: A GTS FWD CVT is rated 23/30 mpg, with a GTS AWD CVT rating 22/29 mpg. Best rating is from the Kizashi S CVT with 23/31 mpg; we're guessing the actual difference is likely a fraction of 1 mpg attributable to the lighter weight and skinnier tires of the base model.
As for the brakes, we found the touch to be beautiful, using them frequently on the curves on the road, and heavily around PIR. We can't imagine anyone in a sports sedan like this one needing more.
The optional all-wheel drive moves the Kizashi into exclusive territory along with the Audi A3 and Subaru Legacy. Called i-AWD, or intelligent all-wheel drive, it's turned on by the driver, and moves up to 50 percent of the torque to the rear wheels."
"The Suzuki Kizashi is a small sports sedan that offers more standard equipment than its competitors, including safety features, at a slightly lower price. The S and SE models offer much, and for more money there are Sport models, plus optional all-wheel drive. The engineering is excellent, with a solid four-cylinder engine, transmission choices including a high-tech CVT with paddle shifters, an exceptionally rigid chassis, tuned suspension offering a compliant ride and great cornering, and precise brakes.
Sam Moses filed this report to NewCarTestDrive.com from Washington's Columbia River Gorge."
Top Cars

subset: commoncrawl


text:
Our Team
Why Weight Loss Surgery?
Weight Loss Procedures
Patient Center
Select a department to call.
October 25, 201910:00 AM
Thank you for choosing Elaine D'Angela for your medical needs.
Insurance plans NOT accepted: Kaiser, Humana, and Medicaid.
Please note the following before scheduling an appointment:
**We currently offer online scheduling with our Nurse Practitioners for 6-month Follow-Up appointments. **We cannot accept online scheduling for surgery at this time. Please contact the office directly at (303) 280-0900. **If you have previously had bariatric surgery and are new to our practice or haven't been seen in over a year, please call us directly to schedule at (303) 280-0900.
If you are unable to find a desired time or need to reschedule an appointment, please contact the office directly at (303) 280-0900 and the receptionist will be happy to accommodate you.
In order to expedite your visit, please bring the following items:
**A list of current medications **Insurance card(s) and a Photo ID
For questions about billing and insurance, please call the office directly at (303) 280-0900.
Thank you again, and we look forward to seeing you!
Denotes required field


text:
The official Bognor RFC app
Bognor RFC Hampshire Avenue Bognor Regis West Sussex PO21 5JH View with Google Maps
This website is part of the Pitchero Rugby Union network. Create your own free Rugby Union club website with Pitchero.
Digital advertising by Minute Media
© Pitch Hero Ltd 2008-2019
Pitch Hero Ltd Registered in ENGLAND | WF3 1DR | Company Number - 636 1033