Validation of Autoclave Protocols for Successful Decontamination of Category A Medical Waste Generated from Care of Patients with Serious Communicable Diseases.

In response to the Ebola outbreak in 2014, many hospitals designated specific areas to care for patients with Ebola and other highly infectious diseases. The safe handling of category A infectious substances is a unique challenge in this environment. One solution is on-site waste treatment with a steam sterilizer or autoclave. The Johns Hopkins Hospital (JHH) installed two pass-through autoclaves in its biocontainment unit (BCU). The JHH BCU and The Johns Hopkins biosafety level 3 (BSL-3) clinical microbiology laboratory designed and validated waste-handling protocols with simulated patient trash to ensure adequate sterilization. The results of the validation process revealed that autoclave factory default settings are potentially ineffective for certain types of medical waste and highlighted the critical role of waste packaging in successful sterilization. The lessons learned from the JHH validation process can inform the design of waste management protocols to ensure effective treatment of highly infectious medical waste.

The Political Effects of Agricultural Subsidies in Africa: Evidence from Malawi.

Across sub-Saharan Africa agricultural subsidy programs have again become a common strategy for combatting rural poverty, increasing agricultural production, and reducing food insecurity. Despite a large literature examining subsidies' effects on output and welfare, little is known about their political effects. This paper examines Malawi's Agricultural Input Subsidy Programme, one of the largest and most expensive programs implemented, which was launched by the government in 2005. We examine whether the incumbent party, the Democratic Progressive Party headed by president Bingu wa Mutharika, benefited from Malawi's subsidy program by examining a longitudinal dataset of 1,846 rural Malawians interviewed in 2008 and again in 2010. The individual-level data show no evidence that the subsidy program was targeted to Mutharika's co-ethnics or co-partisans. Our analysis further demonstrates that the subsidy program increased support for the incumbent party. These results suggest that even when parties are unable or unwilling to target distributional programs at the local level, they may nonetheless derive political benefits. As anti-poverty programs - including agricultural subsidies to small-scale farmers - become increasingly common across the continent, our results suggest that they may help to explain patterns of party affiliation and vote choice, particularly where traditional patterns of partisan affiliation related to ethnic or regional identities weaken.

Perceptions of health professionals on pain in extremely low gestational age infants.

Extremely low gestational age infants (<28 weeks at birth) experience significant pain from repeated therapeutic procedures while hospitalized in the neonatal intensive care unit. As part of a program of research examining pain in preterm infants, we conducted a qualitatively driven mixed-methods design, supplemented with a qualitative and quantitative component, to understand how health care professionals (HCPs) assess and manage procedural pain for tiny and underdeveloped preterm infants. Fifty-nine HCPs from different disciplines across four tertiary-level neonatal units in Canada participated in individual or focus group interviews and completed a brief questionnaire. Four themes from the content analysis were (a) subtlety and unpredictability of pain indicators, (b) infant and caregiver attributes and contextual factors that influence pain response and practices, (c) the complex nature of pain assessment, and (d) uncertainty in the management of pain. The information gleaned from this study can assist in identifying gaps in knowledge and informing unit-based and organizational knowledge translation strategies for this vulnerable population.

Simplified microarray system for simultaneously detecting rifampin, isoniazid, ethambutol, and streptomycin resistance markers in Mycobacterium tuberculosis.

We developed a simplified microarray test for detecting and identifying mutations in rpoB, katG, inhA, embB, and rpsL and compared the analytical performance of the test to that of phenotypic drug susceptibility testing (DST). The analytical sensitivity was estimated to be at least 110 genome copies per amplification reaction. The microarray test correctly detected 95.2% of mutations for which there was a sequence-specific probe on the microarray and 100% of 96 wild-type sequences. In a blinded analysis of 153 clinical isolates, microarray sensitivity for first-line drugs relative to phenotypic DST (true resistance) was 100% for rifampin (RIF) (14/14), 90.0% for isoniazid (INH) (36/40), 70% for ethambutol (EMB) (7/10), and 89.1% (57/64) combined. Microarray specificity (true susceptibility) for first-line agents was 95.0% for RIF (132/139), 98.2% for INH (111/113), and 98.6% for EMB (141/143). Overall microarray specificity for RIF, INH, and EMB combined was 97.2% (384/395). The overall positive and negative predictive values for RIF, INH, and EMB combined were 84.9% and 98.3%, respectively. For the second-line drug streptomycin (STR), overall concordance between the agar proportion method and microarray analysis was 89.5% (137/153). Sensitivity was 34.8% (8/23) because of limited microarray coverage for STR-conferring mutations, and specificity was 99.2% (129/130). All false-susceptible discrepant results were a consequence of DNA mutations that are not represented by a specific microarray probe. There were zero invalid results from 220 total tests. The simplified microarray system is suitable for detecting resistance-conferring mutations in clinical M. tuberculosis isolates and can now be used for prospective trials or integrated into an all-in-one, closed-amplicon consumable.

AIDS exceptionalism: another constituency heard from.

Amidst current debates over resources for AIDS, we examine the policy preferences of the people who are navigating AIDS in their daily lives. Survey and ethnographic data on the prioritization of HIV/AIDS interventions were collected in a longitudinal cohort study in rural Malawi. Study participants gave higher priority to problems other than AIDS. Confining analysis to HIV-positive survey respondents only weakly raises the prioritization of HIV/AIDS services. The ethnographic data are consistent with these findings: although rural Malawians are fully aware of the risk of dying from AIDS, other problems are perceived as more pressing for their community.

Evaluation of the Sensititre MycoTB plate for susceptibility testing of the Mycobacterium tuberculosis complex against first- and second-line agents.

The Sensititre MycoTB plate (TREK Diagnostic Systems, Cleveland, OH) uses a microtiter plate MIC format for susceptibility testing of Mycobacterium tuberculosis complex isolates against first- and second-line antituberculosis agents. Categorical agreement versus the agar proportion method for 122 M. tuberculosis complex isolates was 94% to 100%.

Local Demand for a Global Intervention: Policy Priorities in the Time of AIDS.

- The success of global health and development interventions ultimately depends on local reception. This paper documents local demand for HIV/AIDS interventions in Africa and seeks to explain patterns of demand using data from a country hard-hit by AIDS. As international agencies and national governments scale up HIV/AIDS interventions in Africa, I find HIV-positive respondents more highly prioritize HIV/AIDS programs, however, cross-national opinion data paired with interviews of villagers and their headmen in rural Malawi show weak prioritization of HIV/AIDS. The data illustrate a misalignment of policy preferences in the global-to-local hierarchy, highlighting the import of studying preferences of intended beneficiaries.

Case of Mycobacterium marinum infection with unusual patterns of susceptibility to commonly used antibiotics.

Mycobacterium marinum, found commonly in salt water and freshwater, is the causative agent of disease in many species of fish and occasionally in humans. MICs to most antimicrobial agents are relatively low. Susceptibility testing is not routinely performed, and single-drug therapy is used for the treatment of most infections. Here, we report an infection caused by a drug-resistant M. marinum strain in an otherwise healthy patient.

Evaluation of a rapid fluorescent staining method for detection of mycobacteria in clinical specimens.

Rapid detection of mycobacterial disease is essential. Using multiple specimen types and concentrations of mycobacteria, we compared two commercial auramine O stains. The more rapid stain permitted consistent acid-fast bacillus quantitation and exhibited less debris staining, and the staining procedure required less time ( approximately 2 min) to perform. These results suggest that the rapid stain may be more cost-effective and efficient for use in clinical laboratories.

Automated broth-based systems versus the MYCOTB plate for antimicrobial susceptibility testing of the Mycobacterium tuberculosis complex: challenges in interpretation.

We examined categorical agreement between automated mycobacterial susceptibility testing methods (Mycobacterial Growth Indicator Tube [MGIT] 960 System and the VersaTREK Mycobacteria Detection and Susceptibility System) which are based on single critical concentration (CC) "breakpoints" and a commercial microbroth dilution method (Sensititre Mycobacterium tuberculosis MIC Plate [MYCOTB]) which provides an MIC value. Mycobacterium tuberculosis isolates (n=355) were tested against three first-line antimycobacterial agents (ethambutol [EMB], isoniazid [INH], rifampin [RIF]) using the MYCOTB plate and either the MGIT 960 (site 1, n=142) or VersaTREK (site 2, n=213) systems. Overall categorical agreement was 96.8%. When stratified by drug and CC-defined susceptible and resistant isolates, concordance ranged from 75% to 100%. Interpretation of MIC-based results versus established CC-based results was challenging for drugs whose CC was not represented by an exactly equivalent concentration in the manufacturer-defined dilutions on the MYCOTB plate (EMB, INH). We propose interpretations of MYCOTB plate MICs using the currently available plate configuration.

An unusual antibiotic susceptibility pattern in a Mycobacterium Cosmeticum strain isolated from the chesapeake bay.

OBJECTIVE/BACKGROUND: Mycobacterium cosmeticum, first described in 2004, was recovered from a patient undergoing a cosmetic procedure. Subsequently, this species was associated with an outbreak in a nail salon. In all cases, the isolates were susceptible to all antibiotics tested. Recently, however, we recovered a strain of M. cosmeticum from the Chesapeake Bay, resistant to 11 of 14 antimicrobials. The objective of this work was to present our findings on the resistance and susceptibility of this isolate to various antibiotics. MATERIALS AND METHODS: Surface water samples were collected from 10 sites in the Chesapeake Bay and upper tributaries to assess microbial diversity and antibiotic resistance. Site selection was based on proximity to agricultural runoff, industrial contaminants, and sewage effluents. Samples were processed and recovered organisms were identified and subjected to antimicrobial-susceptibility testing. RESULTS: One nontuberculous species, identified as M. cosmeticum, was recovered from Sandy Point State Park. Resistance was detected to several antibiotics: doxycycline (16 µg/mL), tigecycline (≥4 µg/mL), clarithromycin (8 µg/mL), trimethoprim/sulfamethoxazole (≥8/152 µg/mL), imipenem (32 µg/mL), cefoxitin (32 µg/mL), ethionamide (≥20 µg/mL), and streptomycin (16 µg/mL). Of the 14 antibiotics tested, only the fluoroquinolones, linezolid, and amikacin demonstrated potent activity with susceptible minimum inhibitory concentrations. CONCLUSION: The antimicrobial resistance identified in this M. cosmeticum isolates from the Chesapeake Bay raises some important concerns: (a) why is the susceptibility pattern in this isolate so different from the previously published reports, (b) how did resistance emerge in this isolate,

A rapid, standardized protein extraction method using adaptive focused acoustics for identification of mycobacteria by MALDI-ToF MS.

Mycobacterial identification using MALDI-ToF MS (MALDI) has been hindered by inadequate extraction methods. Adaptive Focused Acoustics™ uses concentrated ultrasonic energy to achieve cellular disruption. Using this technology, we developed a rapid mycobacterial inactivation/protein extraction method for MALDI-based identification. Agreement for identification to the species level versus conventional identification was stratified by log confidence cut-offs of ≥2.0, ≥ 1.8, or ≥1.7. A total of 182 mycobacterial isolates were tested. Complete inactivation of all species/strains was achieved after 2min. Using a log confidence cut-off of ≥2.0, overall agreement for the commercial method (CM) was 41.7% versus 66.7% for the novel method (NM). For the CM, agreement increased to 66.7% and 83.3% using log confidence cut-offs of ≥1.8 and ≥1.7, respectively; for the NM, agreement was 100% for both cut-offs with all isolates. With no alteration to the existing database, overall agreement for the NM was 83.4%, largely due to low scores for clinical isolates of M. chelonae and M. mucogenicum. Addition of spectra from a single clinical strain of each species to the existing database increased overall agreement to 93.1%.

The Creation of a Biocontainment Unit at a Tertiary Care Hospital. The Johns Hopkins Medicine Experience.

In response to the 2014-2015 Ebola virus disease outbreak in West Africa, Johns Hopkins Medicine created a biocontainment unit to care for patients infected with Ebola virus and other high-consequence pathogens. The unit team examined published literature and guidelines, visited two existing U.S. biocontainment units, and contacted national and international experts to inform the design of the physical structure and patient care activities of the unit. The resulting four-bed unit allows for unidirectional flow of providers and materials and has ample space for donning and doffing personal protective equipment. The air-handling system allows treatment of diseases spread by contact, droplet, or airborne routes of transmission. An onsite laboratory and an autoclave waste management system minimize the transport of infectious materials out of the unit. The unit is staffed by self-selected nurses, providers, and support staff with pediatric and adult capabilities. A telecommunications system allows other providers and family members to interact with patients and staff remotely. A full-time nurse educator is responsible for staff training, including quarterly exercises and competency assessment in the donning and doffing of personal protective equipment. The creation of the Johns Hopkins Biocontainment Unit required the highest level of multidisciplinary collaboration. When not used for clinical care and training, the unit will be a site for research and innovation in highly infectious diseases. The lessons learned from the design process can inform a new research agenda focused on the care of patients in a biocontainment environment.

A novel protein extraction method for identification of mycobacteria using MALDI-ToF MS.

Commercial extraction methods for identification of mycobacteria using MALDI-ToF MS are laborious and time consuming. We have developed a novel extraction method which utilizes a bead beater and zirconia/silica beads to significantly shorten the existing protocol. This novel method provides a more rapid extraction of mycobacteria versus the commercial standard.

Mycotic aortic aneurysm due to intravesical BCG immunotherapy: Clinical manifestations and diagnostic challenges.

A live, attenuated form of Mycobacterium bovis, bacillus Calmette-Guérin (BCG), is commonly used as intravesical immunotherapy for non-invasive urothelial bladder carcinoma. While complications are rare, dissemination can occur. A case of mycotic aortic aneurysm following BCG administration with recovery of Mycobacterium bovis in culture is reported. A review of the published experience with this problem is also presented.

Ethnic identity, region and attitudes towards male circumcision in a high HIV-prevalence country.

We study how considerations of male circumcision (MC) as both a favourable practice and as protective against HIV are linked with ethnicity in sub-Saharan Africa, where many ethnic groups do not traditionally circumcise. We focus on Malawi, a country with a high HIV prevalence but low MC prevalence. Survey data from a population-based random sample in rural Malawi (N =3400) were analysed for ethnoregional patterns in attitudes towards MC. We used logit regression models to measure how reported circumcision status, region of residence and ethnic identity relate to attitudes towards circumcision. Overall, Malawians reported more negative than positive opinions about MC, but attitudes towards circumcision varied by ethnicity and region. The implications for agencies and governments aggressively scaling up the provision of MC are clear; acceptance of circumcision as a tool for HIV prevention could be low in societies divided by ethnoregional identities that also shape the practice of circumcision.

An offer you can't refuse? Provider-initiated HIV testing in antenatal clinics in rural Malawi.

International organizations promote provider-initiated, 'routine' HIV testing of pregnant women seeking antenatal care as an effort to curb mother-to-child transmission. We offer an account of the perceptions of HIV testing at antenatal clinics in rural Malawi. Although it is both international and Government of Malawi policy that women must be explicitly informed of their right to refuse testing, analysis of in-depth interviews, focus group discussions and evidence from observational field journals show that rural Malawians do not perceive HIV testing as a choice, but rather as compulsory in order to receive antenatal care. This study illustrates dissonance between global expectations and local realities of the delivery of HIV-testing interventions.

Use of the "RAM" susceptibility testing method for rapid detection of clarithromycin resistance in the Mycobacterium avium complex.

Standard susceptibility testing of the Mycobacterium avium complex (MAC) can require 7 to 14 days from initial isolation. We evaluated a high-performance liquid chromatography-based susceptibility test for rapid determination of clarithromycin (CLR) resistance in MAC. This method can be completed in 72 h of incubation. A total of 110 MAC strains were tested using the following concentrations of CLR: 4, 16, and 64 microg/mL, for a total of 330 tests. Microbroth dilution was used as the reference method. Rapid analysis of mycolic acid ("RAM") concordance with the reference method for CLR susceptibility was 98% (254/258) and 100% for CLR resistance (72/72). The 4 discordant results occurred with 2 strains, which demonstrated intermediate resistance with an MIC of 16 microg/mL. This study demonstrates that "RAM"-based susceptibility testing for determination of CLR resistance in MAC is both rapid and accurate, providing a significant reduction in turn-around-time from 7 to 14 days to 72 h of incubation.

Differences in time to detection and recovery of Mycobacterium spp. between the MGIT 960 and the BacT/ALERT MB automated culture systems.

Mycobacterium sp. recovery and time to detection were compared in the MGIT 960 and BacT/ALERT MB automated broth culture systems. The MGIT 960 demonstrated shorter time to detection (13.5 versus 25.2 days) and greater sensitivity (100% versus 66.6%) for recovery of the Mycobacterium tuberculosis complex than the BacT/ALERT MB system.