Readiness for antimicrobial resistance (AMR) surveillance in Pakistan; a model for laboratory strengthening

Background: Limited capacity of laboratories for antimicrobial susceptibility testing (AST) presents a critical diagnostic bottleneck in resource limited countries. This paper aims to identify such gaps and to explore whether laboratory networks could contribute towards improving AST in low resource settings. Methods: A self-assessment tool to assess antimicrobial susceptibility testing capacity was administered as a pre-workshop activity to participants from 30 microbiology laboratories in 3 cities in Pakistan. Data from public and private laboratories was analyzed and capacity of each scored in percentage terms. Laboratories from Karachi were invited to join a support network. A cohort of five laboratories that consented were provided additional training and updates sessions over a period of 15 months. Impact of training activities in these laboratories was evaluated using a point scoring (0-11) tool. Results: Results of self-assessment component identified a number of areas that required strengthening (scores of ≤60%). These included; readiness for AMR surveillance; 38 and 46%, quality assurance; 49 and 55%, and detection of specific organisms; 56 and 60% for public and private laboratories respectively. No significant difference was detected in AST capacity between public and private laboratories [ANOVA; p \u3e 0.05]. Scoring tool used to assess impact of training within the longitudinal cohort showed an increase from a baseline of 1-5.5 (August 2015) to improved post training scores of 7-11 (October 2016) for the 5 laboratories included. Moreover, statistical analysis using paired t-Test Analysis, assuming unequal variance, indicated that the increase in scored noted represents a statistically significant improvement in the components evaluated [p \u3c 0.05]. Conclusion: Strengthening of laboratory capacity for AMR surveillance is important. Our data shows that close mentoring and support can help enhance capacity for antimicrobial sensitivity testing in resource limited settings. Our study further presents a model wherein laboratory networks can be successfully established and used towards improving diagnostic capacity in such setting

The cytochromes of Acanthamoeba castellanii

1. Low-temperature difference spectra of gradient-purified mitochondria of Acanthamoeba castellanii reveal the presence of cytochromes b-555, b-562 and c-549, with a-type cytochromes having a broad asymmetrical maximum at 602 nm; these components were also observed in specta of whole cells. 2. The a-type cytochromes are unusual in that they have split Soret absorption maxima (at 442 and 449 nm) and an uncharacteristic CO difference spectrum. 3. CO difference spectra of whole cells and 'microsomal' membranes show large amounts of cytochrome P-420 compared with cytochrome P-450. 4. Difference spectra in the presence of cyanide indicate the presence of an a-type cytochrome and two cyanide-reacting components, one of which may be cytochrome a3. 5. Whole-cell respiration in a N2/O2 (19:1) atmosphere was decreased by 50%, suggesting the presence of a low-affinity oxidase. This lowered respiration is inhibited by 50% by CO, and the inhibition is partially light-reversible; photochemical action spectra suggest that cytochrome a3 contributes to this release of inhibition. Other CO-reacting oxidases are also present. 6. The results are discussed with the view that cytochrome a3 is present in A. castellanii, but its identification in CO difference spectra is obscured by other component(s)

Comparative Antifungal Activity of Cilofungin (LY121019) against Candida Species, Including Evaluation of Susceptibility Testing Method

The in vitro activity of cilofungin against 100 Candida species was compared with 5-flucytosine. amphotericin B and ketoconazole by two laboratories independently and in a blinded fashion using a macrotitre dilution broth method in saam-f medium. Cilofungin showed good in vitro activity against Candida albicans. Candida tropicalis and Candida glabrata (90% minimal inhibitory concentration [MIC] 3.2 μg/mL) but was inactive against other Candida species. When testing the susceptibility of cilofungin, 5-flucytosine and amphotericin B at the two centres, approximately 90% of the Candida strains had MICs differing by fourfold or less. However, when testing susceptibility of ketoconazole, only 51% of the Candida strains had MIC differences fourfold or less. MIC susceptibility testing with cilofungin, 5-flucytosine and amphotericin B in saam-f medium is reproducible.Peer Reviewe

Outbreak of Escherichia coli 0157:H7 related to animal contact at a petting zoo

OBJECTIVE: To determine the cause of an outbreak of Escherichia coli 0157:H7 related to animal exposures so that further transmission could be prevented

Additional file 3: of Readiness for antimicrobial resistance (AMR) surveillance in Pakistan; a model for laboratory strengthening

Questionnaire to evaluate impact of knowledge based intervention on laboratory performance during study period. Presents questionnaire used for evaluating impact of knowledge based intervention on laboratory performance during the study period. (DOCX 12 kb

Additional file 2: of Readiness for antimicrobial resistance (AMR) surveillance in Pakistan; a model for laboratory strengthening

Scoring method used including percentage scores for each question in a given category. Total scores and percentage scores for each category of laboratory capacity for both public and private sector are also presented. Presents the scores including total percentage of public and private sector in each category as well as the individual scores in the different components that constituted a particular category. (DOCX 25 kb