Low prevalence of MDR-TB in Lao PDR: results from the first national anti-tuberculosis drug resistance survey.

OBJECTIVE: To present results of the first national anti-tuberculosis (TB) drug resistance survey conducted in Lao PDR between May 2016 and August 2017 to determine the prevalence of resistance to first-line anti-TB drugs among new and previously treated pulmonary TB cases in the country. METHODS: Patients with sputum smear-positive pulmonary TB were enrolled from 42 TB laboratories distributed in 40 clusters throughout the country. Survey sites were selected using probability-proportional-to-size sampling among all diagnostic centres in the country. In addition to smear microscopy, all patients underwent Xpert MTB/RIF testing and those found positive to Mycobacterium tuberculosis underwent sputum culture and drug susceptibility testing using the proportion method on solid Löwenstein-Jensen medium. RESULTS: Among 1006 eligible patients, 946 sputum smear-positive and Xpert MTB/RIF positive (Mycobacterium tuberculosis detected) patients were included in the survey, comprising 897 new and 49 previously treated TB cases. The prevalence of rifampicin-resistant TB was 1.2% (95% CI: 0.5-2.0%, n = 11/897) among new cases and 4.1% (95% CI: 0-9.6%, n = 2/49) among previously treated cases. Among the 946 TB cases confirmed by Xpert MTB/RIF, phenotypic drug sensitivity testing was available for 820 (776 new and 44 previously treated). The prevalence of multidrug-resistant TB (MDR-TB) was 0.5% (95% CI: 0-1.0%, n = 4/776) among new cases and 2.3% (95% CI: 0-6.7%, n = 1/44) among previously treated cases. No resistance to second-line injectable agents nor to fluoroquinolones was detected among MDR-TB patients. CONCLUSIONS: The first national anti-TB drug resistance survey in Lao PDR demonstrated an encouragingly low prevalence of MDR-TB. The results appear lower than previous WHO estimates, and in line with the routine surveillance based on Xpert MTB/RIF testing (conducted among 50% of presumptive TB patients in 2017). The country should continue to expand its Xpert MTB/RIF network and strive to achieve universal drug susceptibility testing.

OBJECTIF: Présenter les résultats de la première surveillance nationale de la résistance aux médicaments antituberculeux, menée en République Démocratique Populaire (RDP) Lao entre mai 2016 et août 2017 afin de déterminer la prévalence de la résistance aux médicaments antituberculeux de première intention chez les nouveaux cas et les cas déjà traités de tuberculose (TB) pulmonaire dans le pays. MÉTHODES: Les patients atteints de TB pulmonaire à frottis d'expectoration positif ont été recrutés dans 42 laboratoires TB répartis dans 40 groupes à travers tout le pays. Les sites de surveillance ont été sélectionnés sur la base d'un échantillon probabiliste proportionnel à la taille parmi tous les centres de diagnostic du pays. Outre l'examen microscopique des frottis, tous les patients ont subi un test Xpert MTB/RIF et ceux trouvés positifs pour Mycobacterium tuberculosis ont subi une culture d'expectorations et un test de sensibilité aux médicaments en utilisant la méthode des proportions sur un milieu solide de Löwenstein-Jensen. RÉSULTATS: Parmi les 1.006 patients éligibles, 946 patients à frottis positif et Xpert MTB/RIF positif (Mycobacterium tuberculosis détecté) ont été inclus dans la surveillance, comprenant 897 nouveaux cas et 49 cas de TB déjà traités. La prévalence de la TB résistante à la rifampicine était de 1,2% (IC95%: 0,5-2,0%, n = 11/897) chez les nouveaux cas et de 4,1% (IC95%: 0-9,6%, n = 2/49) chez les cas traités. Parmi les 946 cas de TB confirmés par Xpert MTB/RIF, des tests de sensibilité phénotypique aux médicaments étaient disponibles pour 820 (776 nouveaux cas et 44 cas traités antérieurement). La prévalence de la TB multirésistante (TB-MDR) était de 0,5% (IC95%: 0-1,0%, n = 4/776) chez les nouveaux cas et de 2,3% (IC95%: 0 à 6,7%, n = 1/44) parmi les cas précédemment traités. Aucune résistance aux agents injectables de deuxième intention ni aux fluoroquinolones n'a été détectée chez les patients atteints de TB-MDR. CONCLUSIONS: La première surveillance nationale de la résistance aux médicaments antituberculeux menée en RDP Lao a révélé une prévalence rassurante de la TB-MDR. Les résultats apparaissent inférieurs aux estimations précédentes de l'OMS et conformes à la surveillance de routine basée sur le test Xpert MTB/RIF (menée auprès de 50% des patients atteints de TB présumée en 2017). Le pays devrait continuer à élargir son réseau Xpert MTB/RIF et s'efforcer d'atteindre des tests universels de sensibilité aux médicaments.

National sentinel surveillance of transmitted drug resistance in antiretroviral-naive chronically HIV-infected patients in France over a decade: 2001-2011.

OBJECTIVES: As recommended by the French ANRS programme for the surveillance of HIV-1 resistance, we estimated the prevalence of transmitted drug resistance-associated mutations (RAMs) in antiretroviral-naive, chronically HIV-1-infected patients. METHODS: RAMs were sought in samples from 661 newly diagnosed HIV-1-infected patients in 2010/11 at 36 HIV clinical care centres. Weighted analyses were used to derive representative estimates of the percentage of patients with RAMs. RESULTS: At patient inclusion, the prevalence of virus with protease (PR) or reverse transcriptase (RT) RAMs was 9.0% (95% CI 6.8%-11.2%). No integrase RAMs were observed. The prevalences of protease inhibitor, nucleoside RT inhibitor and non-nucleoside RT inhibitor RAMs were 1.8%, 6.2% and 2.4%, respectively. Resistance to one, two and three classes of antiretroviral agent was observed in 7.9%, 0.9% and 0.2% of patients, respectively. The frequency of RAMs was higher in patients infected with B compared with non-B subtype virus (11.9% versus 5.1%, P = 0.003). Baseline characteristics (gender, age, country of transmission, CD4 cell count and viral load) were not associated with the prevalence of transmitted RAMs. However, men having sex with men (MSM) were more frequently infected with resistant virus than were other transmission groups (12.5% versus 5.8%, P = 0.003). Compared with the 2006/07 survey, the overall prevalence of resistance remained stable. However, a significant decrease in the frequency of virus with PR RAMs was observed in 2010/11 compared with the 2006/07 survey (1.8% versus 5.0%, P = 0.003). CONCLUSIONS: In France in 2010/11, the global prevalence of transmitted drug-resistant variants was 9.0%, and the prevalence was stable compared with the 2006/07 survey. MSM and B subtype-infected patients are the groups with a higher prevalence of drug resistance.

Target-site resistance to trifluralin is more prevalent in annual ryegrass populations from Western Australia.

BACKGROUND: Trifluralin is widely used in Australia as one of the important pre-emergence herbicides to control annual ryegrass (Lolium rigidum Gaud.) populations. Trifluralin resistance evolution and mechanisms have been identified in some ryegrass populations. RESULTS: In this study, 21 putative resistant field survey populations from Western Australian were screened with trifluralin, and 90% (19 of 21) contained individuals surviving 480 g ha-1 trifluralin treatment. Twelve populations contained individuals possessing the known α-tubulin resistance mutations at Val-202, Thr-239 and Arg-243 in TUA4 (alpha-tubulin 4 n), plus multiple potential resistance mutations in TUA4 pending genetic confirmation. Three populations had only individuals carrying newly identified (but uncharacterized) mutations in TUA3/TUA4. Radioactive work found that six populations evolved metabolic resistance to trifluralin, and at least four of them also possessed the known and/or putative target-site mutations. CONCLUSION: These results confirm that a high incidence of resistance to the dinitroaniline herbicide (trifluralin) is present, and target-site tubulin mutations make a major contribution to resistance in these annual ryegrass populations. Co-evolution of both target-site and non-target-site resistance to per-emergence herbicides warrants diverse management tactics.

Experience on the first national anti-TB drug resistance survey (DRS) in Timor-Leste.

BACKGROUND: A national drug resistance survey (DRS) was implemented for the first time in Timor-Leste (TL) in 2019. The primary objective of the survey was to assess the prevalence of drug resistance among new and previously treated pulmonary TB patients in the country. METHODS: This nation-wide cross-sectional survey was conducted in 2019 targeting all new and previously treated sputum smear-positive pulmonary TB patients. Sputum samples were submitted to the National TB Reference Laboratory for confirmation of TB and to determine resistance to rifampicin by Xpert MTB/RIF. Culture was performed on solid media, and culture isolates of confirmed TB cases were shipped to the WHO Supranational TB Reference Laboratory in Chennai, India for whole genome sequencing (WGS). Survey summary statistics, data cross-tabulations and analysis of potential risk factors of rifampicin-resistant TB (RR-TB) were conducted using R statistical software (version 3.5.2). RESULTS: A total of 953 sputum-smear positive patients were enrolled, of which 917 were confirmed as positive for TB by either Xpert MTB/RIF or culture. An electronic web-based system was used for entry and storage of the data. Rifampicin resistance was detected among 0.6% (95% CI 0.2-1.3) of new cases and 2.7% (95% CI 0.5- 8.2) of previously treated cases. WGS was conducted for validation purposes on 65 randomly selected isolates (29% of RR-TB (2/7) and 7% of RS-TB (63/910) by Xpert MTB/RIF or pDST). The original test results agreed with the WGS validation results for 62/64 isolates (97%). CONCLUSION: The prevalence of RR-TB in Timor-Leste is relatively low compared to the estimated proportions of RR-TB in the WHO South-East Asia Region (2.5% [95% CI 1.9-3.3] among new cases and 14% [95% CI 7.7-21] among previously treated cases). The rapid sputum collection and transportation mechanism implemented in the survey demonstrates its feasibility in low resource settings and should be replicated for routinely transporting TB specimens from microscopy labs to GeneXpert sites. Establishment of in-country capacity for rapid molecular diagnostics for both first- and second-line DST is an immediate need for achieving universal drug susceptibility testing (DST) to guide appropriate patient management.

Herbicide resistance across the Australian continent.

BACKGROUND: Lolium rigidum is the weed of greatest economic impact in Australia due to its formidable capacity to evolve herbicide resistance. In this study, 579 field-sampled L. rigidum populations were tested for resistance to 21 herbicides applied at the recommended rate. Nine herbicide treatments were binary mixtures. RESULTS: A total of 15 876 individual resistance tests were conducted by screening two million seeds at the recommended label rate. The overall frequency of resistant populations was 31%, 14%, 71%, 6% and 0% in response to the post-emergence herbicide treatments clethodim, clethodim + butroxydim, imazamox + imazapyr, glyphosate and paraquat, respectively. The resistance frequency to stand-alone pre-emergence wheat-selective herbicides ranged from 10% to 34%. Conversely, the levels of resistance to pre-emergence mixtures or stand-alone propyzamide were significantly lower, ranging from 6% to 0%. In winter, the responses to glyphosate, paraquat, cinmethylin, prosulfocarb, pyroxasulfone and trifluralin were reassessed, with 7%, 0%, 0%, 21%, 21% and 28% as the respective resistance frequencies. South Australia and Victoria are identified as epicenters for L. rigidum population resistance to pyroxasulfone, whereas populations in New South Wales have the greatest resistance to glyphosate and in Western Australia to clethodim. CONCLUSIONS: For the first time, resistance levels to stand-alone herbicides and binary mixtures are geographically ranked across the Australian continent by benchmark statistical analysis of resistance frequencies and distribution. The extension of these results will raise awareness of rapidly emerging patterns of herbicide resistance, encouraging the adoption of cost-effective modes of action and integration of diverse strategies for weed resistance management.

Area-Wide Survey of Chlorantraniliprole Resistance and Control Failure Likelihood of the Neotropical Coffee Leaf Miner Leucoptera coffeella (Lepidoptera: Lyonetiidae).

The Neotropical coffee leaf miner, Leucoptera coffeella (Guérin-Mèneville & Perrottet, 1842), is a key pest species of unshaded coffee plantations in Neotropical America, particularly in Brazil, where pest management involves intensive insecticide use. As a consequence, problems of resistance to conventional insecticides are frequent, and more recently developed insecticide molecules, such as diamide insecticides, are at risk of becoming ineffective. Thus, a survey of resistance to the diamide insecticide chlorantraniliprole was carried out in high-yield coffee-producing areas in the State of Bahia, Brazil. The likelihood of control failure with this insecticide was also assessed. Spatial dependence among the insect sampling sites was assessed and spatial mapping of chlorantraniliprole resistance and risk of control failure was carried out. The frequency of chlorantraniliprole resistant populations was high (34 out of 40 populations, or 85%), particularly in western Bahia, where 94% of the populations were resistant. Resistance levels ranged from low (<10-fold) to moderate (between 10- and 40-fold) with more serious instances occurring in western Bahia. This results in lower chlorantraniliprole efficacy among these populations, with a higher risk of control failure and exhibiting spatial dependence. These findings invite attention to problems with the intensive use of this relatively recent insecticide and demand management attention, but they suggest that local, farm-based management efforts are likely to be the most effective actions against resistance problems in this pest species.

Insecticide resistance, control failure likelihood and the First Law of Geography.

Insecticide resistance is a broadly recognized ecological backlash resulting from insecticide use and is widely reported among arthropod pest species with well-recognized underlying mechanisms and consequences. Nonetheless, insecticide resistance is the subject of evolving conceptual views that introduces a different concept useful if recognized in its own right - the risk or likelihood of control failure. Here we suggest an experimental approach to assess the likelihood of control failure of an insecticide allowing for consistent decision-making regarding management of insecticide resistance. We also challenge the current emphasis on limited spatial sampling of arthropod populations for resistance diagnosis in favor of comprehensive spatial sampling. This necessarily requires larger population sampling - aiming to use spatial analysis in area-wide surveys - to recognize focal points of insecticide resistance and/or control failure that will better direct management efforts. The continuous geographical scale of such surveys will depend on the arthropod pest species, the pattern of insecticide use and many other potential factors. Regardless, distance dependence among sampling sites should still hold, following the maxim that the closer two things are, the more they resemble each other, which is the basis of Tobler's First Law of Geography. © 2016 Society of Chemical Industry.

Adequacy of examining one sputum specimen in tuberculosis drug resistance surveys.

OBJECTIVE/BACKGROUND: Collection of one spot and one morning sputum specimen is recommended for tuberculosis (TB) drug resistance surveys. This was a retrospective analysis of Mycobacterium tuberculosis cultures isolated from two spot sputum specimens collected from smear positive TB patients in a TB drug resistance survey. It was conducted to understand the value of a second specimen. METHODS: A TB drug resistance survey was conducted in the state of Tamil Nadu, India, to estimate the prevalence of drug resistance among new sputum smear-positive (NSP) and previously treated (PT) patients diagnosed in Revised National Tuberculosis Control Program microscopy centers. A total of 2425 patients (1524 NSP and 901 PT cases) were enrolled in the study. From these patients, two spot sputum specimens (C and D) were collected within a period of 2h. No preservative was added to sputum. The samples were transported at ambient conditions without cold storage to the central laboratory for culture of M. tuberculosis. Culture yield from each sample was computed and analyzed. RESULTS: The proportion of cultures retrieved from C and D specimens among NSP cases (89.3% and 89.7%) and PT cases (90.8% and 90.3%) were similar. The culture grades of C and D samples were comparable (chi-square test, 3560.135; p<.001) and the agreement was moderate (kappa test, 0.454). CONCLUSION: The findings of the study reveal the adequacy of single spot sputum specimen from smear positive pulmonary TB patients for bacteriological examination in a quality-assured TB laboratory to determine precisely the level of drug resistance in a province of India.

Spatial and temporal country-wide survey of temephos resistance in Brazilian populations of Aedes aegypti

The organophosphate temephos has been the main insecticide used against larvae of the dengue and yellow fever mosquito (Aedes aegypti) in Brazil since the mid-1980s. Reports of resistance date back to 1995; however, no systematic reports of widespread temephos resistance have occurred to date. As resistance investigation is paramount for strategic decision-making by health officials, our objective here was to investigate the spatial and temporal spread of temephos resistance in Ae. aegypti in Brazil for the last 12 years using discriminating temephos concentrations and the bioassay protocols of the World Health Organization. The mortality results obtained were subjected to spatial analysis for distance interpolation using semi-variance models to generate maps that depict the spread of temephos resistance in Brazil since 1999. The problem has been expanding. Since 2002-2003, approximately half the country has exhibited mosquito populations resistant to temephos. The frequency of temephos resistance and, likely, control failures, which start when the insecticide mortality level drops below 80%, has increased even further since 2004. Few parts of Brazil are able to achieve the target 80% efficacy threshold by 2010/2011, resulting in a significant risk of control failure by temephos in most of the country. The widespread resistance to temephos in Brazilian Ae. aegypti populations greatly compromise effective mosquito control efforts using this insecticide and indicates the urgent need to identify alternative insecticides aided by the preventive elimination of potential mosquito breeding sites.

Bioassay method for toxicity studies of insecticide formulations to Tuta absoluta (meyrick, 1917) / Metodologia de bioensaio para estudos de toxicidade de formulações comerciais de inseticidas a Tuta absoluta (Meyrick, 1917)

Chemical control is the main method for controlling the tomato leafminer, Tuta absoluta (Meyrick, 1917) (Lepidoptera: Gelechiidae). Reported techniques for the evaluation of insecticide toxicity to the tomato leafminer are not in agreement with field conditions and do not allow us to verify whether doses used in the field are efficient for control. Thus, the objective of this work was to develop a bioassay methodology to study the toxicity of insecticide formulations to T. absoluta that represent field conditions for fast-acting insecticides (neurotoxics and inhibitors of respiration) and slow-acting insecticides (Bacillus thuringiensis and insect growth regulators). The leaf-dip method was the most efficient method for toxicity studies of insecticides formulations to T. absoluta. We verified that bioassays with fast-acting insecticides should be performed with glass Petri dishes containing one tomato foliole from the 4th leaf from the plant apex infested with 10 larvae of 3rd instar and these bioassays can last 48 hours. Conversely, bioassays with slow-acting insecticides should be performed with two-liter transparent PET bottles containing the 4th leaf from the plant apex, with their petioles immersed in a glass bottle containing 120 mL of water, and this leaf should be infested with 10 larvae of 2nd instar and this bioassays can last seven days.

O principal método utilizado no controle da traça-do-tomateiro Tuta absoluta (Meyrick, 1917) (Lepidoptera: Gelechiidae) é a aplicação de inseticidas. As técnicas atuais de avaliação da toxicidade de inseticidas sobre essa praga não simulam a situação de campo e não possibilitam a verificação se as doses usadas no campo são eficientes no seu controle. Assim, neste trabalho, objetivou-se desenvolver uma metodologia que represente as condições de campo para inseticidas de ação rápida (neurotóxicos e inibidores respiratórios) e de ação lenta (Bacillus thuringiensis e reguladores de crescimento. A metodologia mais eficiente para estudos de toxicidade de formulações comerciais a T. absoluta foi a imersão de folhas em calda inseticida. Para os bioensaios de inseticidas de ação rápida, sugere-se que estes sejam realizados em placas de Petri, contendo folíolos de tomate da 4ª folha a partir do ápice da planta, infestados com 10 larvas de 3º ínstar e eles podem durar 48 horas. Quanto aos bioensaios de toxicidade de inseticidas de ação lenta, sugere-se que sejam realizados em garrafas PET transparentes, de dois litros, contendo a 4ª folha de tomate a partir do ápice da planta infestada com 10 larvas de 2º ínstar e seu pecíolo inserido em vidro de 120 mL contendo água. Nesse caso, o bioensaio pode durar sete dias sem prejuízo na eficiência.