Predictive profiling of gram-negative antibiotics in CagA oncoprotein inactivation: a molecular dynamics simulation approach.

Gastric cancer (GC) is the fifth most prevalent form of cancer worldwide. CagA - positive Helicobacter pylori infects more than 60% of the human population. Moreover, chronic infection of CagA-positive H. pylori can directly affect GC incidence. In the current study, we have repurposed FDA-approved antibiotics that are viable alternatives to current regimens and can potentially be used as combination therapy against the CagA of H. pylori. The 100 FDA-approved gram negative antibiotics were screened against CagA protein using the AutoDock 4.2 tool. Further, top nine compounds were selected based on higher binding affinity with CagA. The trajectory analysis of MD simulations reflected that binding of these drugs with CagA stabilizes the system. Nonetheless, atomic density map and principal component analysis also support the notion of stable binding of antibiotics to the protein. The residues ASP96, GLN100, PRO184, and THR185 of compound cefpiramide, doxycycline, delafloxacin, metacycline, oxytetracycline, and ertapenem were involved in the binding with CagA protein. These residues are crucial for the CagA that aids in entry or pathogenesis of the bacterium. The screened FDA-approved antibiotics have a potential druggability to inhibit CagA and reduce the progression of H. pylori borne diseases.

Selective photoionization of lithium isotopes in a hollow cathode lamp: a feasibility study for a laser ion source and detector.

Two-step selective photoionization of lithium isotopes is carried out in a hollow cathode (HC) discharge lamp using a nitrogen-laser-pumped homemade tunable dye laser (∼0.2 cm-1, 7 ns, 640-680 nm). The HC lamp is developed in-house and used as a lithium ion source as well as a detector. The dark space region of HC discharge is illuminated by a dye laser exciting pulse followed by an ionizing radiation from the nitrogen laser. The high electric field (∼kV/cm) present in HC dark space is exploited for fast charge collection, produced by the simultaneous interaction of exciting and ionizing laser beams with sputtered lithium atoms, enabling the detection of photoionization signal. Photoionization signals with a high signal-to-noise ratio are obtained without using any amplifier. Effects of the ionizing laser pulse energy and the precise tuning of the dye laser on lithium photoionization signal, across 670.780 nm (S1/22→P1/2,3/22) transition, are studied. The fine structure of lithium is clearly identified, and its two isotopes (Li7,6) are fairly resolved in the measured photoionization spectrum. Using a saturation technique, the photoionization cross section and the number density of lithium atoms for the Li7 (P3/22) excited level are also measured as 18.5±2.4 Mb, N0∼1.7×109 (atoms/cm3). The results obtained infer that HC-lamp-based photoionization is an easier and cost-effective way of selective photoionization of lithium atoms, and it can be applied for other elements also.

Development of a see-through hollow cathode discharge lamp for (Li/Ne) optogalvanic studies.

Development of a demountable and see-through hollow cathode (HC) discharge lamp suitable for optogalvanic (OG) spectroscopy is described. The design of the HC lamp is simple, compact, and inexpensive. Lithium, investigated rarely by the OG method, is selected for cathode material as its isotopes are important for nuclear industry. The HC lamp is characterized electrically and optically for discharge oscillations free OG effect. Strong OG signals of lithium as well as neon (as buffer gas) are produced precisely upon copper vapor laser pumped tunable dye laser irradiation. The HC lamp is capable of generating a clean OG resonance spectrum in the available dye laser wavelength scanning range (627.5-676 nm) obtained with 4-(Dicyanomethylene)-2-methyl-6-(4-dimethylaminostyryl)-4H-pyran dye. About 28 resonant OG lines are explicitly observed. Majority of them have been identified using j-l coupling scheme and assigned to the well-known neon transitions. One line that corresponds to wavelength near about 670.80 nm is assigned to lithium and resolved for its fine (2S1/2 → 2P1/2, 3/2) transitions. These OG transitions allow 0.33 cm-1 accuracy and can be used to supplement the OG transition data available from other sources to calibrate the wavelength of a scanning dye laser with precision at atomic levels.

Hyperfine structure studies of neutral europium transitions at 601.815 and 580.027 nm by saturation absorption spectroscopy.

We report Doppler-free spectroscopy of neutral europium carried out using the saturation absorption technique. In the present experiment, Eu/Ne hollow cathode discharge is used as an atomic vapor source of europium. The Eu transition 4f76s2(S87/2°)→4f76s6p(P89/2) at 601.815 nm and 4f75d6s(aD1013/2°)→4f75d6p(zD811/2) at 580.027 nm are investigated. The frequency separation between the adjacent hyperfine transitions as well as hyperfine structure (HFS) constants are calculated from the obtained experimental spectra. The HFS constant obtained for the state 4f76s6p(P89/2) are A151=664.8 MHz; B151=293.5 MHz, and for state 4f75d6p(zD811/2), the values are A151=7.6 MHz; B151=-60 MHz. These obtained values are in good agreement with the results reported in the literature using laser induced atomic beam fluorescence technique.

NEED OF IRON SUPPLEMENTATION IN GESTATIONAL DIABETES MELLITUS.

OBJECTIVE: Gestational diabetes mellitus (GDM) increases the foetal morbidity and mortality and there is also risk of development of type II diabetes for the mother. In pregnancy iron supplementations are given to all the females in India. Data are not available on the relationship between serum ferritin and GDM in Indian populations. MATERIAL & METHODS: Case control study was done on 90 women (30 GDM patients approx 28 weeks of gestation and 60 control subjects without GDM) referred to the tertiary care hospital at New Delhi. Ferritin was analysed by ELISA kit. They were analyzed for the correlation of ferritin with blood glucose in GDM patients. RESULTS: On comparison with blood glucose the levels of ferritin were statistically significantly high in GDM cases as compared to controls (p value 0.008). There was a negative correlation of ferritin with blood glucose level (r value -0.039, p value 0.794). DISCUSSION: Pregnancy is an equivalent of an inflammatory state and the high levels of ferritin can help in determining the need of iron supplementation given in pregnant females. Women who had high serum ferritin had the greatest risk of GDM. CONCLUSION: This study suggests a possible link between elevated serum ferritin and low-grade inflammation. Further studies in larger populations are required to establish this study.

Studies on laser-assisted Penning ionization by the optogalvanic effect in Ne/Eu hollow cathode discharge.

Laser-assisted Penning ionization (LAPI) is detected in a Ne/Eu hollow cathode (HC) discharge lamp using the pulsed optogalvanic (OG) method. In the Ne/Eu discharge, doubly ionized europium excited energy levels Eu[4f(7)(P(7/2,5/2)6)] lie within the thermal limit (∼kT) from the laser-excited neon's energy level [2p(5)(P3/202)3p or 2p(8) (in Paschen notation)] lying at 149,848 cm(-1). Therefore, Penning ionization (PI) of europium atoms likely to occur into its highly excited ionic states is investigated. To probe the PI of europium, the temporal profiles of its counterpart neon OG signal are studied as a function of discharge current for the transitions (1s(4)→2p(8)) and (1s(2)→2p(2)), corresponding to 650.65 and 659.89 nm wavelengths, respectively. It is observed that PI of europium alters the overall discharge characteristics significantly and, hence, modifies the temporal profile of the OG signals accordingly. The quasi-resonant ionizing energy transfer collisions between laser-excited Ne 2p(8) atoms and electronically excited europium P(9/2)10 atoms are used to explain the LAPI mechanism. Such LAPI studies carried out in HC discharge could be useful for the discharge of a metal-vapor laser with appropriate Penning mixtures.

Prospective Observational Study of Adverse Drug Reactions of Anticancer Drugs Used in Cancer Treatment in a Tertiary Care Hospital.

Adverse drug reactions associated with the use of anticancer drugs are a worldwide problem and cannot be ignored. Adverse drug reactions can range from nausea, vomiting or any other mild reaction to severe myelosuppression. The study was planned to observe the suspected adverse drug reactions of cancer chemotherapy in patients aged >18 years having cancer attending Postgraduate Institute of Medical Education and Research, Chandigarh. During the study period, 101 patients of breast cancer and 73 patients of lung cancer were screened for occurrence of adverse drug reactions during their treatment with chemotherapy. About 87.36% patients experienced adverse drug reactions, 90.09% and 83.56% of breast and lung cancer patients experienced at least one adverse drug reaction respectively. In breast cancer patients, 41.58% patients were prescribed fluorouracil+doxorubicin+cyclophosphamide while paclitaxel was prescribed to 22.77% patients. Alopecia (54.94%), nail discolouration (43.96%), dysgeusia (38.46%), anorexia (30.77%), nausea (29.67%), and neuropathy (29.67%) were found to be very common in breast cancer patients treated with single/combined regimen. In lung cancer group of patients, cisplatin with docetaxel, cisplatin with pemetrexed and cisplatin with irinotecan were prescribed to 30.14, 24.65 and 17.81% patients, respectively. Dysgeusia (40.98%), diarrhoea (39.34%), anorexia (32.77%) and constipation (31.15%) and alopecia (31.15%) were commonly observed adverse drug reactions having lung cancer patients. Causality assessments using World Health Organization causality assessment scale showed that observed adverse drug reactions were of probable (64.67%) and possible (35.33%) categories. Alopecia, dysgeusia, anorexia, constipation diarrhoea, nausea, nail discoloration were more prevalent amongst the cancer patients undergoing chemotherapy.

Advances in diabetic retinopathy.

Diabetic retinopathy (DR) is a complication of long-term diabetes mellitus (DM). Over the last 2 decades lot of work has been on early diagnosis of DR and screening programs have been designed to help the masses. Large numbers of clinical studies have been done for patients of diabetes and DR wherein the role of blood sugar control, metabolic control, role of oral medicines for DR, role of imaging, fluorescein angiography, and retinal photocoagulation has been studied. Newer treatment modalities are being devised and studied for better patient care. We discuss these issues in our review highlight and newer advances over the last few years.

QSAR analyses of DDT analogues and their in silico validation using molecular docking study against voltage-gated sodium channel of Anopheles funestus.

DDT has enjoyed the reputation of a successful pesticide in disease control programme and agricultural practices along with the serious opposition and ban later on due to its biomagnification and toxic action against non-target organisms. The present work was carried out to develop QSAR models for analysing DDT analogues for their pesticidal activity and in silico validation of these models. A 2D-QSAR model was generated using stepwise with multiple regression, and the model with a value of r(2) = 0.7324; q(2) = 0.6215; pred r(2) = 0.7038, containing five descriptors, was selected for further study. The 3D QSAR with CoMFA analysis showed that steric contribution of 21% and electrostatic contribution of about 79% were required for larvicidal activity of DDT analogues. A set of 3430 molecules was generated using the basic DDT skeleton as template, and these were evaluated for their mosquito larvicidal activity using the generated QSAR models and DDT as standard. Eleven molecules were selected for in silico validation of these models. For this, a docking study of the selected molecules against the homology-modelled voltage-gated sodium channel of Anopheles funestus was conducted. The study showed that the activities of these analogues as predicted by 2D-QSAR, 3D-QSAR with CoMFA and dock score were observed to be well correlated.

Studies on pulsed optogalvanic effect in Eu/Ne hollow cathode discharge.

The optogalvanic (OG) effect has been observed in a Eu/Ne hollow cathode discharge lamp using pulsed laser irradiation. An OG spectrum is recorded in dye laser wavelength region 574­602 nm using a boxcar-averager. In total 41 atomic lines are observed. Of these, 38 lines are assigned to neon transitions. Two lines observed corresponding to wavelengths 576.519 and 601.815 nm are assigned to europium transitions; (4f 7 6s 2 , S 8 7/2 →4f 7 6s6p , zP 6 7/2 ) and (4f 7 6s 2 , S 8 7/2 →4f 7 6s6p , zP 8 9/2 ), respectively, and the remaining line at 582.475 nm could not be assigned. The effect of the discharge current on europium as well as neon OG signals is also studied. At moderate discharge current values, an extra positive peak is observed in neon OG signal for the transition (1s 5 →2p 2 ) at 588.189 nm, which is explained by Penning-ionization process using the quasi-resonant energy transfer interactions between excited neon and europium atoms lying in 2p 2 and D 10 9/2 states, respectively.

Knowledge and use of personal protective measures against mosquito borne diseases in a resettlement colony of delhi.

BACKGROUND: Mosquito borne diseases (MBDs) are major public health problem in India. State of Delhi is endemic for dengue and other MBDs. The increasing incidence of MBDs in Delhi in recent years warrants a pro-active approach for their prevention. Knowledge and use of personal protective measures (PPMs) presents an effective strategy for prevention and control of MBDs. AIM: The present study was conducted to assess the knowledge and use of PPMs against MBDs in an urban resettlement colony of Delhi. SUBJECTS AND METHODS: It was a cross-sectional study carried out in a resettlement colony of Delhi. A total of 100 families were selected by systematic random sampling. Data was collected using semi-structured questionnaire and supplemented by spot survey by the investigator in the community. The results were analyzed in SPSS version 16.0 (Chicago Illinios, USA). RESULTS: Out of the 100 respondents, 65% (65/100), 58%(58/100) and 13% (13/100) had heard about dengue, malaria and chikungunya, respectively. Nearly, one-fifth (20/100; 20%) of the participants reported incorrect breeding sites for mosquitoes. The knowledge regarding PPMs was very high (93/100; 93%) and about (90/100; 90%) families were actually using at least one of the PPMs. However, very few families were using them correctly (1/90; 1.1%) and adequately (5/90; 5.6%). The most common PPM being used by the study population was liquid vaporizers (54/90; 60%). Nearly one-third (29/90; 32.2%) of the participants reported side-effects due to PPMs with irritation to smell being the most common reported side-effect. On house visit, adult mosquitoes were seen in 67% (67/100) of the houses, while potential mosquito breeding sites were found in and around 56% (56/100) houses. CONCLUSIONS: There were crucial gaps in knowledge and practices of participants with regard to prevention and control of MBDs. Thus, there is a need to intensify efforts toward creating public knowledge and mobilizing community about correct use of preventive measures against MBDs.

Macular Thickness Variability in Primary Open Angle Glaucoma Patients using Optical Coherence Tomography.

AIM: To compare the difference of retinal macular thickness and macular volume using optical coherence tomography (OCT) in primary open angle glaucoma (POAG) patients with the normal subjects. MATERIALS AND METHODS: This observational case control study included primary open angle glaucoma (POAG) patients (n = 124 eyes) and healthy subjects in the control group (n = 124 eyes). All subjects underwent detailed history, general and systemic exami -nation. Complete ocular examination included best corrected visual acuity (BCVA), slit lamp examination, intraocular pressure (IOP), central corneal thickness, gonioscopy, dilated fundus biomicroscopy. Field analysis was done by white on white Humphrey Field Analyzer (Carl Zeiss). Optical coherence tomography imaging of macular area was performed using Stratus OCT (OCT 3, Version 4, Carl Zeiss Inc, Dublin, California, USA). In both these groups, parameters analyzed were macular thickness, inner macular thicknesses (IMT), outer macular thicknesses (OMT), central macular thick ness (CMT) and total macular volume (TMV). RESULTS: The POAG group had significantly decreased values of TMV, OMT and IMT, compared to control group, while there was no difference in CMT, presumably due to absence of ganglion cells in the central part. Thus, macular thickness and volume parameters may be used for making the diagnosis of glaucoma especially in patients with abnormalities of disc. CONCLUSION: Macular thickness parameters correlated well with the diagnosis of glaucoma. How to cite this article: Sharma A, Agarwal P, Sathyan P, Saini VK. Macular Thickness Variability in Primary Open Angle Glaucoma Patients using Optical Coherence Tomography. J Current Glau Prac 2014;8(1):10-14.

Evaluation of Central Macular Thickness and Retinal Nerve Fiber Layer Thickness using Spectral Domain Optical Coherence Tomography in a Tertiary Care Hospital.

PURPOSE: To evaluate the normative data of macular thickness and retinal nerve fiber layer thickness (RNFL) among normal subjects using spectral domain optical coherence tomography (OCT). MATERIALS AND METHODS: Normal subjects presenting to a tertiary medical hospital were included in the study. All patient underwent clinical examination followed by study of macular thickness and RN FL thick ness by spectral domain Topc on OCT. The data was collected and analyzed for variations in gender and age. The data was also compared with available literature. RESULTS: Total numbers of patients enrolled in the study were 154 (308 eyes). Numbers of males were 79 (158 eyes) and numbers of females were 75 (150 eyes). The mean age among males was 42.67 ± 12.15 years and mean age among females was 42.88 ± 11.73 years. Overall the mean mac ular thickness (central 1 mm zone) with SD - OCT was 241.75 ± 17.3 microns. The mean macular volume was 7.6 cu. mm ± 0.33. On analysis of the RNFL thickness, we observed that the RNFL was thickest in the inferior quadrant (138.58) followed by superior (122.30) nasal (116.32) and temporal quadrant (73.04). Gender-wise comparison of the data revealed no statistically significant difference for age, macular thickness parameters, volume and RFNL values except outer temporal thickness among males and females. No age-related difference was noted in the above parameters. On comparison with available norma tive data from India and elsewhere, we found significant variations with different machines. CONCLUSION: The study is the first to provide normative data using SD-OCT from central India. The data from spectral domain OCT correlated well with the values obtained from similar studies with SD - OCT. Values obtained from time domain OCT machines are different and are not comparable. How to cite this article: Agarwal P, Saini VK, Gupta S, Sharma A. Evaluation of Central Macular Thickness and Retinal Nerve Fiber Layer Thickness using Spectral Domain Optical Coherence Tomography in a Tertiary Care Hospital. J Curr Glaucoma Pract 2014;8(2):75-81.

Laser-induced optogalvanic signal oscillations in miniature neon glow discharge plasma.

Laser-induced optogalvanic (OG) signal oscillations detected in miniature neon glow discharge plasma are investigated using a discharge equivalent-circuit model. The damped oscillations in OG signal are generated when a pulsed dye laser is tuned to a specific neon transition (1s5→2p2) at 588.2 nm under the discharge conditions where dynamic resistance changes its sign. Penning ionization via quasi-resonant energy transfer collisions between neon gas atoms in metastable state and sputtered electrode atoms in ground state is discussed to explain the negative differential resistance properties of discharge plasma that are attributed to oscillations in the OG signal. The experimentally observed results are simulated by analyzing the behavior of an equivalent discharge-OG circuit. Good agreement between theoretically calculated and experimental results is observed. It is found that discharge plasma is more sensitive and less stable in close vicinity to dynamic resistance sign inversion, which can be useful for weak-optical-transition OG detection.

Antimicrobial resistance profiles of common mastitis pathogens on Canadian dairy farms.

Monitoring of antimicrobial resistance (AMR) in bacteria has clinical and public health significance. The present study determined prevalence of AMR in common mastitis pathogens Staphylococcus aureus, including methicillin-resistant Staph. aureus (MRSA; n=1,810), Escherichia coli (n=394), and Klebsiella species (n=139), including extended-spectrum ß-lactamase (ESBL)-producing E. coli and Klebsiella species, isolated from milk samples on 89 dairy farms in 6 Canadian provinces. Minimum inhibitory concentrations (MIC) were determined using the Sensititer bovine mastitis plate (Trek Diagnostic Systems Inc., Cleveland, OH) and a National Antimicrobial Resistance Monitoring System gram-negative panel containing antimicrobials commonly used for mastitis treatment and control. Denim blue chromogenic agar and real-time PCR were used to screen and confirm MRSA, respectively. Resistance proportion estimates ranged from 0% for cephalothin and oxacillin to 8.8% for penicillin in Staph. aureus isolates, and 15% of the resistant Staph. aureus isolates were multidrug resistant. One MRSA isolate was confirmed (prevalence: 0.05%). Resistance proportion estimates ranged from 0% for ceftriaxone and ciprofloxacin to 14.8% for tetracycline in E. coli, and 0% for amikacin, ceftiofur, ciprofloxacin, and nalidixic acid to 18.6% for tetracycline in Klebsiella species isolates. Further, 62.8 and 55% of the resistant E. coli and Klebsiella species isolates were multidrug resistant, respectively. Resistance to >5 and >2 antimicrobials was most common in E. coli and Klebsiella species isolates, respectively, and no ESBL producers were found. Prevalence of AMR in bovine mastitis pathogens was low. Most gram-negative udder pathogens were multidrug resistant; MRSA was rarely found, and ESBL E. coli and Klebsiella species isolates were absent in Canadian milk samples.

Herd-level association between antimicrobial use and antimicrobial resistance in bovine mastitis Staphylococcus aureus isolates on Canadian dairy farms.

Surveillance of antimicrobial use and resistance is needed to manage antimicrobial resistance in bacteria. In this study, data were collected on antimicrobial use and resistance in Staphylococcus aureus (n=562), isolated from intramammary infections and (sub)clinical mastitis cases on 89 dairy farms in 4 regions of Canada [Alberta, Ontario, Québec, and the Maritime Provinces (Prince Edward Island, Nova Scotia, and New Brunswick)]. Dairy producers were asked to deposit empty drug containers into specially provided receptacles, and antimicrobial drug use rate was calculated to quantify antimicrobial use. Minimum inhibitory concentrations were determined using the Sensititer bovine mastitis plate system (TREK Diagnostic Systems Inc., Cleveland, OH), containing antimicrobials commonly used for mastitis treatment and control. Multivariable logistic regression models were built to determine herd-level risk factors of penicillin, ampicillin, pirlimycin, penicillin-novobiocin combination, tetracycline and sulfadimethoxine resistance in Staph. aureus isolates. Intramammary administration of the penicillin-novobiocin combination for dry cow therapy was associated with penicillin and ampicillin resistance [odds ratio (OR): 2.17 and 3.10, respectively]. Systemic administration of penicillin was associated with penicillin resistance (OR: 1.63). Intramammary administration of pirlimycin for lactating cow mastitis treatment was associated with pirlimycin resistance as well (OR: 2.07). Average herd parity was associated with ampicillin and tetracycline resistance (OR: 3.88 and 0.02, respectively). Average herd size was also associated with tetracycline resistance (OR: 1.02). Dairy herds in the Maritime region had higher odds of penicillin and lower odds of ampicillin resistance than dairy herds in Québec (OR: 2.18 and 0.19, respectively). Alberta dairy herds had lower odds of ampicillin and sulfadimethoxine resistance than dairy herds in Québec (OR: 0.04 and 0.08, respectively). Ontario dairy herds had lower odds of tetracycline and sulfadimethoxine resistance than dairy herds in Québec (OR: 0.05 and 0.33, respectively). Herd-level use of certain antimicrobials administered for mastitis treatment and control, such as intramammary penicillin and pirlimycin as well as systemically administered penicillin and florfenicol, was positively associated with antimicrobial resistance in bovine mastitis pathogens in the field conditions. Differences in antimicrobial resistance outcomes across 4 regions of Canada were observed.

Antimicrobial use on Canadian dairy farms.

Antimicrobial use (AMU) data are critical for formulating policies for containing antimicrobial resistance. The present study determined AMU on Canadian dairy farms and characterized variation in AMU based on herd-level factors such as milk production, somatic cell count, herd size, geographic region and housing type. Drug use data were collected on 89 dairy herds in 4 regions of Canada, Alberta, Ontario, Québec, and the Maritime provinces (Prince Edward Island, New Brunswick, and Nova Scotia) for an average of 540 d per herd. Dairy producers and farm personnel were asked to deposit empty drug containers into specially provided receptacles. Antimicrobial use was measured as antimicrobial drug use rate (ADUR), with the unit being number of animal defined-daily doses (ADD)/1,000 cow-days. Antimicrobial drug use rates were determined at farm, region, and national level. Combined ADUR of all antimicrobial classes was 14.35 ADD/1,000 cow-days nationally. National level ADUR of the 6 most commonly used antimicrobial drug classes, cephalosporins, penicillins, penicillin combinations, tetracyclines, trimethoprim-sulfonamide combinations, and lincosamides were 3.05, 2.56, 2.20, 1.83, 0.87, and 0.84 ADD/1,000 cow-days, respectively. Dairy herds in Ontario were higher users of third-generation cephalosporins (ceftiofur) than in Québec. Alberta dairy herds were higher users of tetracyclines in comparison to Maritimes. Antimicrobial drug use rate was higher via systemic route as compared with intramammary and other routes of administration (topical, oral, and intrauterine). The ADUR of antimicrobials used intramammarily was higher for clinical mastitis treatment than dry cow therapy. For dry cow therapy, penicillin ADUR was greater than ADUR of first-generation cephalosporins. For clinical mastitis treatment, ADUR of intramammary penicillin combinations was greater than ADUR of cephapirin. Herd-level milk production was positively associated with overall ADUR, ADUR of systemically administered ceftiofur, cephapirin administered for dry cow therapy, and pirlimycin administered for clinical mastitis treatment. Herd size and ADUR of systemically administered ceftiofur were also positively associated. In conclusion, ß-lactams were most commonly used on Canadian dairy farms. Among antimicrobials of very high importance in human medicine, the use of fluoroquinolones was rare, whereas third-generation cephalosporins and penicillin combinations containing colistin were used very frequently on Canadian dairy farms.

Diagnostic accuracy assessment of Sensititre and agar disk diffusion for determining antimicrobial resistance profiles of bovine clinical mastitis pathogens.

Determining the accuracy and precision of a measuring instrument is pertinent in antimicrobial susceptibility testing. This study was conducted to predict the diagnostic accuracy of the Sensititre MIC mastitis panel (Sensititre) and agar disk diffusion (ADD) method with reference to the manual broth microdilution test method for antimicrobial resistance profiling of Escherichia coli (n = 156), Staphylococcus aureus (n = 154), streptococcal (n = 116), and enterococcal (n = 31) bovine clinical mastitis isolates. The activities of ampicillin, ceftiofur, cephalothin, erythromycin, oxacillin, penicillin, the penicillin-novobiocin combination, pirlimycin, and tetracycline were tested against the isolates. Diagnostic accuracy was determined by estimating the area under the receiver operating characteristic curve; intertest essential and categorical agreements were determined as well. Sensititre and the ADD method demonstrated moderate to highly accurate (71 to 99%) and moderate to perfect (71 to 100%) predictive accuracies for 74 and 76% of the isolate-antimicrobial MIC combinations, respectively. However, the diagnostic accuracy was low for S. aureus-ceftiofur/oxacillin combinations and other streptococcus-ampicillin combinations by either testing method. Essential agreement between Sensititre automatic MIC readings and MIC readings obtained by the broth microdilution test method was 87%. Essential agreement between Sensititre automatic and manual MIC reading methods was 97%. Furthermore, the ADD test method and Sensititre MIC method exhibited 92 and 91% categorical agreement (sensitive, intermediate, resistant) of results, respectively, compared with the reference method. However, both methods demonstrated lower agreement for E. coli-ampicillin/cephalothin combinations than for Gram-positive isolates. In conclusion, the Sensititre and ADD methods had moderate to high diagnostic accuracy and very good essential and categorical agreement for most udder pathogen-antimicrobial combinations and can be readily employed in veterinary diagnostic laboratories.