Sources of Multidrug Resistance in Patients With Previous Isoniazid-Resistant Tuberculosis Identified Using Whole Genome Sequencing: A Longitudinal Cohort Study

Background Meta-analysis of patients with isoniazid-resistant tuberculosis given standard first-line anti-tuberculosis treatment indicated an increased risk of multi-drug resistant tuberculosis (MDR-TB) emerging (8%), compared to drug-sensitive tuberculosis (0.3%). Here we use whole genome sequencing (WGS) to investigate whether treatment of patients with pre-existing isoniazid resistant disease with first-line anti-tuberculosis therapy risks selecting for rifampicin resistance, and hence MDR-TB. Methods Patients with isoniazid-resistant pulmonary TB were recruited and followed up for 24 months. Drug-susceptibility testing was performed by Microscopic observation drug-susceptibility assay (MODS), Mycobacterial Growth Indicator Tube (MGIT) and by WGS on isolates at first presentation and in the case of re-presentation. Where MDR-TB was diagnosed, WGS was used to determine the genomic relatedness between initial and subsequent isolates. De novo emergence of MDR-TB was assumed where the genomic distance was five or fewer single nucleotide polymorphisms (SNPs) whereas reinfection with a different MDR-TB strain was assumed where the distance was 10 or more SNPs. Results 239 patients with isoniazid-resistant pulmonary tuberculosis were recruited. Fourteen (14/239, 5.9%) patients were diagnosed with a second episode of tuberculosis that was multi-drug resistant. Six (6/239, 2.5%) were identified as having evolved MDR-TB de novo and six as having been re-infected with a different strain. In two cases the genomic distance was between 5-10 SNPs and therefore indeterminate. Conclusions In isoniazid-resistant TB, de novo emergence and reinfection of MDR-TB strains equally contributed to MDR development. Early diagnosis and optimal treatment of isoniazid resistant TB are urgently needed to avert the de novo emergence of MDR-TB during treatment

A prospective multi-center observational study of children hospitalized with diarrhea in Ho Chi Minh City, Vietnam.

We performed a prospective multicenter study to address the lack of data on the etiology, clinical and demographic features of hospitalized pediatric diarrhea in Ho Chi Minh City (HCMC), Vietnam. Over 2,000 (1,419 symptomatic and 609 non-diarrheal control) children were enrolled in three hospitals over a 1-year period in 2009-2010. Aiming to detect a panel of pathogens, we identified a known diarrheal pathogen in stool samples from 1,067/1,419 (75.2%) children with diarrhea and from 81/609 (13.3%) children without diarrhea. Rotavirus predominated in the symptomatic children (664/1,419; 46.8%), followed by norovirus (293/1,419; 20.6%). The bacterial pathogens Salmonella, Campylobacter, and Shigella were cumulatively isolated from 204/1,419 (14.4%) diarrheal children and exhibited extensive antimicrobial resistance, most notably to fluoroquinolones and third-generation cephalosporins. We suggest renewed efforts in generation and implementation of policies to control the sale and prescription of antimicrobials to curb bacterial resistance and advise consideration of a subsidized rotavirus vaccination policy to limit the morbidity due to diarrheal disease in Vietnam

Virulence of Mycobacterium tuberculosis Clinical Isolates Is Associated With Sputum Pre-treatment Bacterial Load, Lineage, Survival in Macrophages, and Cytokine Response

It is uncertain whether differences in Mycobacterium tuberculosis (Mtb) virulence defined in vitro influence clinical tuberculosis pathogenesis, transmission, and mortality. We primarily used a macrophage lysis model to characterize the virulence of Mtb isolates collected from 153 Vietnamese adults with pulmonary tuberculosis. The virulence phenotypes were then investigated for their relationship with sputum bacterial load, bacterial lineages, bacterial growth, and cytokine responses in macrophages. Over 6 days of infection, 34 isolates (22.2%) showed low virulence (< 5% macrophages lysed), 46 isolates (30.1%) showed high virulence (≥90% lysis of macrophages), and 73 isolates (47.7%) were of intermediate virulence (5–90% macrophages lysed). Highly virulent isolates were associated with an increased bacterial load in patients' sputum before anti-tuberculosis therapy (P = 0.02). Isolate-dependent virulence phenotype was consistent in both THP-1 and human monocyte-derived macrophages. High virulence isolates survived better and replicated in macrophages one hundred fold faster than those with low virulence. Macrophages infected with high virulence isolates produced lower concentrations of TNF-α and IL-6 (P = 0.002 and 0.0005, respectively), but higher concentration of IL-1β (P = 5.1 × 10−5) compared to those infected with low virulence isolates. High virulence was strongly associated with East Asian/Beijing lineage [P = 0.002, Odd ratio (OR) = 4.32, 95% confident intervals (CI) 1.68–11.13]. The association between virulence phenotypes, bacterial growth, and proinflammatory cytokines in macrophages suggest the suppression of certain proinflammatory cytokines (TNF-α and IL-6) but not IL-1β allows better intracellular survival of highly virulent Mtb. This could result in rapid macrophage lysis and higher bacterial load in sputum of patients infected with high virulence isolates, which may contribute to the pathogenesis and success of the Beijing lineage

Awareness and preparedness of healthcare workers against the first wave of the COVID-19 pandemic: A cross-sectional survey across 57 countries.

BACKGROUND: Since the COVID-19 pandemic began, there have been concerns related to the preparedness of healthcare workers (HCWs). This study aimed to describe the level of awareness and preparedness of hospital HCWs at the time of the first wave. METHODS: This multinational, multicenter, cross-sectional survey was conducted among hospital HCWs from February to May 2020. We used a hierarchical logistic regression multivariate analysis to adjust the influence of variables based on awareness and preparedness. We then used association rule mining to identify relationships between HCW confidence in handling suspected COVID-19 patients and prior COVID-19 case-management training. RESULTS: We surveyed 24,653 HCWs from 371 hospitals across 57 countries and received 17,302 responses from 70.2% HCWs overall. The median COVID-19 preparedness score was 11.0 (interquartile range [IQR] = 6.0-14.0) and the median awareness score was 29.6 (IQR = 26.6-32.6). HCWs at COVID-19 designated facilities with previous outbreak experience, or HCWs who were trained for dealing with the SARS-CoV-2 outbreak, had significantly higher levels of preparedness and awareness (p<0.001). Association rule mining suggests that nurses and doctors who had a 'great-extent-of-confidence' in handling suspected COVID-19 patients had participated in COVID-19 training courses. Male participants (mean difference = 0.34; 95% CI = 0.22, 0.46; p<0.001) and nurses (mean difference = 0.67; 95% CI = 0.53, 0.81; p<0.001) had higher preparedness scores compared to women participants and doctors. INTERPRETATION: There was an unsurprising high level of awareness and preparedness among HCWs who participated in COVID-19 training courses. However, disparity existed along the lines of gender and type of HCW. It is unknown whether the difference in COVID-19 preparedness that we detected early in the pandemic may have translated into disproportionate SARS-CoV-2 burden of disease by gender or HCW type

A hidden HIV epidemic among women in Vietnam

<p>Abstract</p> <p>Background</p> <p>The HIV epidemic in Vietnam is still concentrated among high risk populations, including IDU and FSW. The response of the government has focused on the recognized high risk populations, mainly young male drug users. This concentration on one high risk population may leave other populations under-protected or unprepared for the risk and the consequences of HIV infection. In particular, attention to women's risks of exposure and needs for care may not receive sufficient attention as long as the perception persists that the epidemic is predominantly among young males. Without more knowledge of the epidemic among women, policy makers and planners cannot ensure that programs will also serve women's needs.</p> <p>Methods</p> <p>More than 300 documents appearing in the period 1990 to 2005 were gathered and reviewed to build an understanding of HIV infection and related risk behaviors among women and of the changes over time that may suggest needed policy changes.</p> <p>Results</p> <p>It appears that the risk of HIV transmission among women in Vietnam has been underestimated; the reported data may represent as little as 16% of the real number. Although modeling predicted that there would be 98,500 cases of HIV-infected women in 2005, only 15,633 were accounted for in reports from the health system. That could mean that in 2005, up to 83,000 women infected with HIV have not been detected by the health care system, for a number of possible reasons. For both detection and prevention, these women can be divided into sub-groups with different risk characteristics. They can be infected by sharing needles and syringes with IDU partners, or by having unsafe sex with clients, husbands or lovers. However, most new infections among women can be traced to sexual relations with young male injecting drug users engaged in extramarital sex. Each of these groups may need different interventions to increase the detection rate and thus ensure that the women receive the care they need.</p> <p>Conclusion</p> <p>Women in Vietnam are increasingly at risk of HIV transmission but that risk is under-reported and under-recognized. The reasons are that women are not getting tested, are not aware of risks, do not protect themselves and are not being protected by men. Based on this information, policy-makers and planners can develop better prevention and care programs that not only address women's needs but also reduce further spread of the infection among the general population.</p

Influence of Stress and Antibiotic Resistance on Cell-Length Distribution in Mycobacterium tuberculosis Clinical Isolates

Mycobacterial cellular variations in growth and division increase heterogeneity in cell length, possibly contributing to cell-to-cell variation in host and antibiotic stress tolerance. This may be one of the factors influencing Mycobacterium tuberculosis persistence to antibiotics. Tuberculosis (TB) is a major public health problem in developing countries, antibiotic persistence, and emergence of antibiotic resistance further complicates this problem. We wanted to investigate the factors influencing cell-length distribution in clinical M. tuberculosis strains. In parallel we examined M. tuberculosis cell-length distribution in a large set of clinical strains (n = 158) from ex vivo sputum samples, in vitro macrophage models, and in vitro cultures. Our aim was to understand the influence of clinically relevant factors such as host stresses, M. tuberculosis lineages, antibiotic resistance, antibiotic concentrations, and disease severity on the cell size distribution in clinical M. tuberculosis strains. Increased cell size and cell-to-cell variation in cell length were associated with bacteria in sputum and infected macrophages rather than liquid culture. Multidrug-resistant (MDR) strains displayed increased cell length heterogeneity compared to sensitive strains in infected macrophages and also during growth under rifampicin (RIF) treatment. Importantly, increased cell length was also associated with pulmonary TB disease severity. Supporting these findings, individual host stresses, such as oxidative stress and iron deficiency, increased cell-length heterogeneity of M. tuberculosis strains. In addition we also observed synergism between host stress and RIF treatment in increasing cell length in MDR-TB strains. This study has identified some clinical factors contributing to cell-length heterogeneity in clinical M. tuberculosis strains. The role of these cellular adaptations to host and antibiotic tolerance needs further investigation

Comparative Examination of Phytonutrients and Antioxidant Activity of Commonly Consumed Nuts and Seeds Grown in Vietnam

The aim of the present study was to determine the phenolics, carotenoids, B-vitamins, and antioxidant activity of nuts and seeds grown in Vietnam. The concentrations of carotenoids and B-vitamins may vary among the nuts and seeds. Watermelon seed contained the highest level of lutein while pumpkin seed was the β-carotene richest sample. Sachi inchi and sunflower seed comprised considerable levels of vitamin B1, B6, and B9. The phenolic analysis revealed that cashew contained the highest total amount of flavonoids (466.04 μg/g), with catechin, epicatechin, and procyanidin B2 predominating over the other flavonoids. Likewise, chlorogenic and neochlorogenic acids made up the highest total amount of phenolic acids in sunflower seed (1870.41 μg/g). Walnut appeared to possess the highest antioxidant activity evaluated by DPPH, ABTS, FRAP, and reducing power assays. The correlation analysis indicated strong positive correlations between total phenolic content with DPPH and FRAP values. Principal component analysis graphically showed the distant positioning of cashew and sunflower seed, highlighting their significantly higher levels of phenolics. The findings of the study would be useful to improve nutrient database contents for flavonoids and phenolic acids as well as to promote the consumption of nut and seed products in Vietnam

Viral load testing to monitor the HIV epidemic among PWID in Vietnam

Objective: To share Vietnam’s experiences piloting the integration of viral load (VL) testing into the national HIV sentinel surveillance (HSS) system to better understand the level of HIV viral transmission among people who inject drugs (PWID).Introduction: Vietnam initiated the HSS system in 1994 in selected provinces with high HIV burden. The surveillance has two components: monitor HIV sero-prevalence and risk behaviors among key population including PWID. However, no VL data were collected among HIV infected people. In 2016, Vietnam piloted an added component of VL testing to the existing HSS system. The purpose was to test the feasibility of adding VL testing to the HSS so that VL data among PWID would be available. The pilot was conducted in two provinces in southern Vietnam-Ho Chi Minh City and Long An. It was expected that adding the VL testing to the existing HSS would also save resources and help monitor HIV viral transmission among PWID in the community regardless if they are currently on anti-retroviral therapy (ART).Methods: Male PWIDs were enrolled into 2016 HSS+ following the standard operating procedure (SOP)[1]. Community-based sampling was based on random selection of wards/communes listed in the sampling frame. In each selected ward/commune, all eligible PWID were invited to voluntarily participate in the survey. Eligibility criteria were males 16 years of age or older, reporting injecting drug in the past month, and residing in the selected area. . The survey included an interview using a standardized questionnaire and 7ml blood drawn for HIV testing. Blood specimens were transferred from districts to provincial labs for plasma separation in the same day. Each plasma specimen was divided into three aliquots of 1ml each. One aliquot was used to test for HIV diagnosis at provincial labs, using the national HIV testing strategy III[2]. The remaining 2 aliquots were stored at provincial labs at 2-80C and within 5 days, were shipped to Pasteur Institute in Ho Chi Minh City (PIHCM) where the plasma specimens were stored at -800C. Processing of samples for VL testing was conducted at the end of the survey where all plasma specimen were transferred to PIHCM lab, which was 2 months since the collection of the first blood specimen. VL was undertaken on COBAS AMPLYPREP/COBAS TAQMAN 48, with identification threshold 20 cps/ml and specificity of 100% using Kit CAP-G/CTM HIV-1 V 2.0. The VL testing results were sent back to relevant Provicial AIDS Centers to return to respective participants, within 3 months.Results: Five hundred male PWID (HCMC: 300; LA: 200) were enrolled into 2016 HSS/HSS+ and agreed to provide blood specimen without any refusal. 84 tested positive for HIV (16.8%. HCMC: 15.0%; LA: 19.5%), 43 (51.2%) specimens had unsuppressed VL (&gt;1000 copies/ml) (HCMC: 66.7%; LA: 33.3%), 35 (41.7%) specimens had undetected level (&lt;50 copies/ml or undetected) (HCMC: 31.1%; LA: 53.9%), and 7.1% had VL that ranged from 50-1000 copies/ml (HCMC: 2.2%; LA: 12.8%). Among those who had VL &lt; 1000 copies/ml, 22 (53.7%) had ever been on ART.Conclusions: The pilot survey has measured VL among male PWID, including those who were aware of their HIV status and those who did not know their status before. Findings indicate that a significant proportion of PWID do not have their VL suppressed leading to high-risk of HIV transmission from PWID to their sexual partners[3] in the community although level of unsuppressed viral load is not a direct measure of HIV viral transmission in itself. This pilot indicated that it was feasible to add VL testing into HSS and Vietnam government can add it as a routine practice in HSS and can be expanded in the coming years