Hepatitis parasitaria por Lamanema chavezi en guanacos (Lama guanicoe) faenados en la Provincia de Santa Cruz, Argentina

Durante la zafra 2017-18 en un frigorífico de la localidad de Río Gallegos, Argentina, se detectaron lesiones en hígados de guanacos (Lama guanicoe) a la faena. Con el fin de caracterizar y determinar la causa de las lesiones se tomaron muestras de tejidos para histopatología y de materia fecal para análisis coproparasitológicos de 10 animales. Macroscópicamente se observaron nódulos esféricos, blanco-amarillentos, firmes al tacto de diversos tamaños. Estas lesiones estaban demarcadas por una cápsula fibrosa con infiltración de neutrófilos y macrófagos. Además, se detectaron lesiones más pequeñas, sin cápsula fibrosa, algunas hemorrágicas y otras con infiltrado celular linfocitario. En los cortes histológicos del hígado de un guanaco se observaron secciones de parásitos. En el examen coproparasitológico se observaron huevos de Lamanema chavezi, Nematodirus spp., Capillaria spp. y Trichuris spp. No se observaron huevos de Fasciola hepatica. Los coprocultivos confirmaron el desarrollo de larvas de 680-800 μm, con colas con terminación roma y presencia de ocho células intestinales coincidentes con lo descripto para L. chavezi. Se concluye que las lesiones hepáticas observadas se deben a la migración de las formas larvarias de L. chavezi, siendo este el primer reporte de esta patología en poblaciones silvestres de guanacos en la Patagonia Argentina.During the 2017-18 season in a slaughterhouse in Rio Gallegos, Argentina, macroscopic lesions in the liver of wild guanacos (Lama guanicoe) were detected. With the aim to characterize and to determine de cause of the lesions, tissues samples for histopathological studies and faeces for copro-parasitological analysis were collected from 10 animals. Macroscopically, spherical, various sizes, white-yellowish of firm consistency nodules were detected. These lesions were demarcated by a fibrous capsule with a cellular infiltration including degenerated neutrophils and macrophages. In addition, smaller focal lesions were also detected, without fibrous capsule, some haemorrhagic and others with mainly lymphocyte infiltration. In liver sections of one animal, several structures of parasites were observed. Eggs of Lamanema chavezi, Nematodirus spp., Capillaria spp. and Trichuris spp. were observed during the parasitological examination of faeces. No eggs of Fasciola hepatica were detected. It is concluded that the lesions observed were due to the migration of the larval forms of L. chavezi, being this the first report of this pathology in wild populations of guanacos in the Argentine Patagonia.Estación Experimental Agropecuaria Bariloche. Área Producción Animal. Grupo Sanidad AnimalFil: Santana, Jorge Luis. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Santa Cruz. Agencia de Extensión Rural Rio Gallegos; ArgentinaFil: Martinez, Agustin. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Bariloche. Área Producción Animal. Grupo Sanidad Animal; ArgentinaFil: Soulés, Anabel. Servicio Nacional de Sanidad Animal. Área de Inocuidad y Calidad Agroalimentaria; ArgentinaFil: Milicevic, Francisco. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Santa Cruz. Agencia de Extensión Rural Rio Gallegos; ArgentinaFil: Cafrune Wierna, María Mercedes. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Salta. Área Salud Animal; ArgentinaFil: Larroza, Marcela Patricia. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Bariloche. Área Producción Animal. Grupo Sanidad Animal; ArgentinaFil: Robles, Carlos Alejandro. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Bariloche. Area de Produccion Animal. Grupo de Sanidad Animal; Argentin

Correlations of mutations in katG, oxyR-ahpC and inhA genes and in vitro susceptibility in Mycobacterium tuberculosis clinical strains segregated by spoligotype families from tuberculosis prevalent countries in South America

Background Mutations associated with resistance to rifampin or streptomycin have been reported for W/Beijing and Latin American Mediterranean (LAM) strain families of Mycobacterium tuberculosis. A few studies with limited sample sizes have separately evaluated mutations in katG, ahpC and inhA genes that are associated with isoniazid (INH) resistance. Increasing prevalence of INH resistance, especially in high tuberculosis (TB) prevalent countries is worsening the burden of TB control programs, since similar transmission rates are noted for INH susceptible and resistant M. tuberculosis strains. Results We, therefore, conducted a comprehensive evaluation of INH resistant M. tuberculosis strains (n = 224) from three South American countries with high burden of drug resistant TB to characterize mutations in katG, ahpC and inhA gene loci and correlate with minimal inhibitory concentrations (MIC) levels and spoligotype strain family. Mutations in katG were observed in 181 (80.8%) of the isolates of which 178 (98.3%) was contributed by the katG S315T mutation. Additional mutations seen included oxyR-ahpC; inhA regulatory region and inhA structural gene. The S315T katG mutation was significantly more likely to be associated with MIC for INH ≥2 μg/mL. The S315T katG mutation was also more frequent in Haarlem family strains than LAM (n = 81) and T strain families. Conclusion Our data suggests that genetic screening for the S315T katG mutation may provide rapid information for anti-TB regimen selection, epidemiological monitoring of INH resistance and, possibly, to track transmission of INH resistant strains.Fil: Dalla Costa, Elis R. State Foundation for Production and Research in Health (FEPPS); Brasil.Fil: Ribeiro, Marta O. State Foundation for Production and Research in Health (FEPPS); Brasil.Fil: Silva, Márcia S. N. State Foundation for Production and Research in Health (FEPPS); Brasil.Fil: Arnold, Liane S. State Foundation for Production and Research in Health (FEPPS); Brasil.Fil: Rostirolla, Diana C. State Foundation for Production and Research in Health (FEPPS); Brasil.Fil: Cafrune, Patricia I. State Foundation for Production and Research in Health (FEPPS); Brasil.Fil: Espinoza, Roger C. Blufstein Clinic Laboratory; Perú.Fil: Palaci, Moises. Federal University of Espírito Santo; Brasil.Fil: Telles, Maria A. Adolfo Lutz Institute; Brasil.Fil: Ritacco, Viviana. ANLIS Dr.C.G.Malbrán. Instituto Nacional de Enfermedades Infecciosas. Servicio de Micobacterias; Argentina.Fil: Suffys, Philip N. Oswaldo Cruz Institute; Brasil.Fil: Lopes, Maria L. Evandro Chagas Institute; Brasil.Fil: Campelo, Creuza L. LACEN Ceará; BrasilFil: Miranda, Silvana S. Federal University of Minas Gerais; Brasil.Fil: Kremer, Kristin. National Institute for Public Healthand the Environment (RIVM). Mycobacteria Reference Unit (CIb-LIS); Países Bajos.Fil: Almeida da Silva, Pedro E. Federal Foundation of Rio Grande; Brasil.Fil: de Souza Fonseca, Leila. Federal University of Rio de Janeiro. Tuberculosis Academic Program; Brasil.Fil: Ho, John L. Cornell University; Estados Unidos.Fil: Kritski, Afrânio L. Federal University of Rio de Janeiro. Tuberculosis Academic Program; Brasil.Fil: Rossetti, María L. R. State Foundation for Production and Research in Health (FEPPS); Brasil

Avaliação da técnica de spoligotyping aplicada diretamente em amostras clínicas de pacientes com suspeita de tuberculose e caracterização epidemiológica

A tuberculose (TB) permanece como uma doença importante do ponto de vista da saúde pública. Devido ao aumento do número de casos, surgimento de cepas resistentes aos fármacos e a co-infecção pelo HIV, novas estratégias para combater a doença são urgentemente necessárias. Por isso a Organização Mundial da Saúde recomenda investimentos em novas tecnologias e estratégias que possam contribuir para o diagnóstico mais rápido, assim como a aplicação de ferramentas que permitam um melhor entendimento da epidemiologia da TB em diferentes populações. O spoligotyping é uma técnica desenvolvida para detecção e diferenciação simultânea de cepas do complexo Mycobacterium tuberculosis. A oportunidade de combinar informações diagnósticas rápidas e dados de epidemiologia molecular representa um avanço importante no controle da TB. Dados epidemiológicos, caracterização molecular a partir de amostras clínicas e geolocalização dos casos de TB, podem contribuir para o entendimento da distribuição da doença e auxiliar no desenvolvimento de estratégias mais específicas para conter sua disseminação. Para isso foi realizado um estudo prospectivo incluindo pacientes atendidos no ambulatório do Hospital Sanatório Partenon, de agosto de 2005 a abril de 2007. Foram incluídos 202 pacientes, 142 com TB e 60 sem a doença. Entre os pacientes com TB a média de idade foi mais baixa (36,96±12,17 vs 44,54±6,05), com maior frequência eram não-brancos, desempregados, sem renda, utilizavam drogas, tinham histórico de encarceramneto, eram desnutridos e tinham tido contato com paciente com TB, principalmete intradomiciliar e em prisões. Os pacientes não-brancos apresentaram maiores taxas de infecção por HIV e HCV. Alcoolismo estava associado ao abandono do tratamento. A técnica de spoligotyping apresentou um bom rendimento quando aplicada diretamente em amostras clínicas, uma vez que 89% das amostras apresentaram resultados concordantes. Entre os 135 isolados de pacientes genotipados, foram identificados 44 spoligotipos diferentes, sendo 112 isolados (83%) com padrões compartilhados e 23 (17%) com padrões únicos. A família LAM foi a mais frequente englobando 37% dos isolados. Através da geolocalização foi possível observar que há uma grande diversidade genética das cepas circulantes na região estudada, mas também alguns agrupamentos que devem ser melhor caracterizados. A estratégia proposta neste estudo, utilizar o spoligotyping diretamente em amostras clínicas, mostrou-se útil para encurtar o tempo de obtenção de informações sobre as identidades das cepas de M. tuberculosis. Pode ser utilizada em combinação com dados clínicos dos pacientes para gerar informações epidemiológicas muito mais rapidamente e auxiliar no combate à TB.Tuberculosis (TB) remains as an important public helath issue. Because of the increase in the number of cases, emergence of drug resistant strains and HIV coinfection, new strategies to fight the disease are urgently needed. Thus, the World Health Organization recommends investiments on new approaches and technologies that can be used to fasten the diagnosis of TB, as well as the application of tools that allow a better understanding of the epidemiology of the disease in different populations. Spoligotyping is a technique which was developed for simultaneous detection and differentiation of Mycobacterium tuberculosis complex strains. The opportunity to combine rapid diagnostic information and molecular epidemiology represents an important advance in the epidemiologic control of TB. Epidemiologic data, molecular characterization from clinical samples and spatial localization of TB cases may be used for the understanding of disease distribution and can be useful for the development of more specific strategies to arrest its dissemination. Aiming at this, a prospective study was designed including patients assisted at the outpatient section of Hospital Sanatório Partenon from August 2005 to April 2007. It was included 202 patients from wich 142 had a positive diagnosis for TB and 60 had a negative diagnosis for the disease. Mean age was lower among patients diagnosed with TB (36.96±12.17 vs 44.54±6.05), they were more likely to be non-white, to be unemployed and not to have any income, to use drugs, to have incarceration history, to have malnutrition and have had contact with a patiente with TB, mainly in the household and in prisons. Non-white patients had higher rates of HIV and HCV co-infection. Alcohol abuse was associated to treatment nonadherence. Spoligotyping technique presented good results when applied directly to clinical samples, once 89% of samples presented agreeing results. Among 135 patients isolates genotyped, it was found 44 differnt spoligotypes, 112 (83%) were shared types and 23(17%) were unique. LAM family was the most frequent comprising 37% of all isolates. Spatial localization of cases demonstrated a high genetic diversity of strains circulating in the study area, but also pointed some clusters that should be better characterized. The strategy proposed in this study, applying spoligotyping directly to clinical samples, showed to be useful to shorten time in obtaining information on M. tuberculosis strains identities. It can be used in combination with clinical data of patients to yield epidemiologic inofrmation more rapidly and help to fight TB

Comparison of two laboratory-developed PCR methods for the diagnosis of pulmonary tuberculosis in Brazilian patients with and without HIV infection

Background: Direct smear examination with Ziehl-Neelsen (ZN) staining for the diagnosis of pulmonary tuberculosis (PTB) is cheap and easy to use, but its low sensitivity is a major drawback, particularly in HIV seropositive patients. As such, new tools for laboratory diagnosis are urgently needed to improve the case detection rate, especially in regions with a high prevalence of TB and HIV. Objective: To evaluate the performance of two in house PCR (Polymerase Chain Reaction): PCR dot-blot methodology (PCR dot-blot) and PCR agarose gel electrophoresis (PCR-AG) for the diagnosis of Pulmonary Tuberculosis (PTB) in HIV seropositive and HIV seronegative patients. Methods: A prospective study was conducted (from May 2003 to May 2004) in a TB/HIV reference hospital. Sputum specimens from 277 PTB suspects were tested by Acid Fast Bacilli (AFB) smear, Culture and in house PCR assays (PCR dot-blot and PCR-AG) and their performances evaluated. Positive cultures combined with the definition of clinical pulmonary TB were employed as the gold standard. Results: The overall prevalence of PTB was 46% (128/277); in HIV+, prevalence was 54.0% (40/74). The sensitivity and specificity of PCR dot-blot were 74% (CI 95%; 66.1%-81.2%) and 85% (CI 95%; 78.8%-90.3%); and of PCR-AG were 43% (CI 95%; 34.5%-51.6%) and 76% (CI 95%; 69.2%-82.8%), respectively. For HIV seropositive and HIV seronegative samples, sensitivities of PCR dot-blot (72% vs 75%; p = 0.46) and PCR-AG (42% vs 43%; p = 0.54) were similar. Among HIV seronegative patients and PTB suspects, ROC analysis presented the following values for the AFB smear (0.837), Culture (0.926), PCR dot-blot (0.801) and PCR-AG (0.599). In HIV seropositive patients, these area values were (0.713), (0.900), (0.789) and (0.595), respectively. Conclusion: Results of this study demonstrate that the in house PCR dot blot may be an improvement for ruling out PTB diagnosis in PTB suspects assisted at hospitals with a high prevalence of TB/HIV

La correspondencia de Valencia : diario de noticias : eco imparcial de la opinión y de la prensa: Año XXXIV Número 15232 - 1911 Noviembre 27

Copia digital. Madrid : Ministerio de Cultura. Subdirección General de Coordinación Bibliotecaria, 200

Comparação das técnicas de double repetitive element pcr (DRE-PCR) e restriction fragment length polymorphism (RFLP) para a identificação de linhagens de Mycobacterium tuberculosis

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Comparison of two laboratory-developed PCR methods for the diagnosis of pulmonary tuberculosis in Brazilian patients with and without HIV infection

Background: Direct smear examination with Ziehl-Neelsen (ZN) staining for the diagnosis of pulmonary tuberculosis (PTB) is cheap and easy to use, but its low sensitivity is a major drawback, particularly in HIV seropositive patients. As such, new tools for laboratory diagnosis are urgently needed to improve the case detection rate, especially in regions with a high prevalence of TB and HIV. Objective: To evaluate the performance of two in house PCR (Polymerase Chain Reaction): PCR dot-blot methodology (PCR dot-blot) and PCR agarose gel electrophoresis (PCR-AG) for the diagnosis of Pulmonary Tuberculosis (PTB) in HIV seropositive and HIV seronegative patients. Methods: A prospective study was conducted (from May 2003 to May 2004) in a TB/HIV reference hospital. Sputum specimens from 277 PTB suspects were tested by Acid Fast Bacilli (AFB) smear, Culture and in house PCR assays (PCR dot-blot and PCR-AG) and their performances evaluated. Positive cultures combined with the definition of clinical pulmonary TB were employed as the gold standard. Results: The overall prevalence of PTB was 46% (128/277); in HIV+, prevalence was 54.0% (40/74). The sensitivity and specificity of PCR dot-blot were 74% (CI 95%; 66.1%-81.2%) and 85% (CI 95%; 78.8%-90.3%); and of PCR-AG were 43% (CI 95%; 34.5%-51.6%) and 76% (CI 95%; 69.2%-82.8%), respectively. For HIV seropositive and HIV seronegative samples, sensitivities of PCR dot-blot (72% vs 75%; p = 0.46) and PCR-AG (42% vs 43%; p = 0.54) were similar. Among HIV seronegative patients and PTB suspects, ROC analysis presented the following values for the AFB smear (0.837), Culture (0.926), PCR dot-blot (0.801) and PCR-AG (0.599). In HIV seropositive patients, these area values were (0.713), (0.900), (0.789) and (0.595), respectively. Conclusion: Results of this study demonstrate that the in house PCR dot blot may be an improvement for ruling out PTB diagnosis in PTB suspects assisted at hospitals with a high prevalence of TB/HIV

Colorimetric microwell plate reverse-hybridization assay for Mycobacterium tuberculosis detection

Direct smear examination using Ziehl-Neelsen staining for pulmonary tuberculosis (PTB) diagnosis is inexpensive and easy to use, but has the major limitation of low sensitivity. Rapid molecular methods are becoming more widely available in centralized laboratories, but they depend on timely reporting of results and strict quality assurance obtainable only from costly commercial kits available in high burden nations. This study describes a pre-commercial colorimetric method, Detect-TB, for detecting Mycobacterium tuberculosis DNA in which an oligonucleotide probe is fixed onto wells of microwell plates and hybridized with biotinylated polymerase chain reaction amplification products derived from clinical samples. The probe is capable of hybridising with the IS6110 insertion element and was used to specifically recognise the M. tuberculosis complex. When combined with an improved silica-based DNA extraction method, the sensitivity of the test was 50 colony-forming units of the M. tuberculosis reference strain H37Rv. The results that were in agreement with reference detection methods were observed in 95.2% (453/476) of samples included in the analysis. Sensitivity and specificity for 301 induced sputum samples and 175 spontaneous sputum samples were 85% and 98%, and 94% and 100%, respectively. This colorimetric method showed similar specificity to that described for commercially available kits and may provide an important contribution for PTB diagnosis