Exploring Differences in Culturable Fungal Diversity Using Standard Freezing Incubation-A Case Study in the Limestones of Lemos Pantheon (Portugal).

In this study, we explored the biodiversity and abundance of culturable fungi in four samples associated with different biodeterioration outlines collected from the Lemos Pantheon, a limestone-built artwork in Portugal. We compared the results from prolonged standard freezing with those previously obtained from fresh samples to analyze differences in the obtained community and assess the effectiveness of the standard freezing incubation protocol in uncovering a different segment of culturable fungal diversity. Our results showed a slight decrease in culturable diversity, but over 70% of the obtained isolates were not present in the previously studied fresh samples. We also identified a high number of potential new species with this procedure. Moreover, the use of a wide variety of selective culture media positively influenced the diversity of the cultivable fungi obtained in this study. These findings highlight the importance of developing new protocols under varying conditions to accurately characterize the culturable fraction in a given sample. The identification and study of these communities and their possible contribution to the biodeterioration process is crucial knowledge for formulating effective conservation and restoration plans to prevent further damage to valuable cultural heritage assets.

Leprosy decline in Amazonas State, Brazil.

OBJECTIVE: To analyse the leprosy case detection rates in Amazonas State, Brazil, by age group from 1980 to 2009. METHOD: The historical data series of leprosy cases by age group from 1980 to 2009 were fitted as a function of time using Poisson regression models. Relative annual reduction in the detection rate (RAR) by age group was estimated as one minus the exponential of the estimated regression coefficient for time. To compare the regression coefficients, we used their 95% confidence interval. RESULTS: The relative annual reduction varied from 9% in the age group of 0-4 years to 1% in the age group of 60-69 years. There was a declining trend of the RAR in the younger age groups that disappeared after 29 years of age. The detection rate in people >29 years old declined very little over time, with no statistically significant difference between age groups. CONCLUSION: Our findings show a reduction in the infection risk in the last 30 years and a birth cohort effect: cohorts born in more recent years faced smaller risks of leprosy infection than older cohorts.

Leprosy survey among rural communities and wild armadillos from Amazonas state, Northern Brazil.

There is evidence that in southern US, leprosy is a zoonosis infecting wild Dasypus novemcinctus armadillos but the extent of this finding is unknown. This ecological study investigated leprosy in rural communities and in wild armadillos from the Brazilian Amazon. The study area was the Mamiá Lake of Coari municipality, Amazonas State, Northern region, a hyper endemic leprosy area where residents live on subsistence farming, fishing and armadillo hunting and its meat intake are frequent. The leprosy survey was conducted in sixteen communities by a visiting team of specialists. Local partakers provided wild armadillos to investigate M. leprae infection. Volunteers had complete dermato-neurological examination by a dermatologist with expertise in leprosy diagnosis, suspect skin lesions were biopsied for histopathology (Hematoxylin-eosin/HE, Fite-Faraco/FF staining); slit skin smears were collected. Armadillos' tissue fragments (skins, spleens, livers, lymph nodes, adrenal glands, others) were prepared for histopathology (HE/FF) and for M. leprae repetitive element-RLEP-qPCR. Among 176 volunteers, six new indeterminate leprosy cases were identified (incidence = 3.4%). Suspect skin sections and slit skin smears were negative for bacilli. Twelve wild D. novemcinctus were investigated (48 specimens/96 slides) and histopathological features of M. leprae infection were not found, except for one skin presenting unspecific inflammatory infiltrate suggestive of indeterminate leprosy. Possible traumatic neuroma, granuloma with epithelioid and Langhans cells, foreign-body granuloma were also identified. Granulomatous/non-granulomatous dermatitides were periodic-acid-Schiff/PAS negative for fungus. M. leprae-RLEP-qPCR was negative in all armadillos' tissues; no bacillus was found in histopathology. Our survey in rural communities confirmed the high endemicity for leprosy while one armadillo was compatible with paucibacillary M. leprae infection. At least in the highly endemic rural area of Coari, in the Brazilian Amazon region where infectious sources from untreated multibacillary leprosy are abundant, M. leprae infected armadillos may not represent a major source of infection nor a significant public health concern.

[Leprosy in indigenous populations of Amazonas State, Brazil: an epidemiological study in the counties of Autazes, Eirunepé and São Gabriel da Cachoeira (2000 to 2005)]. / Hanseníase em populações indígenas do Amazonas, Brasil: um estudo epidemiológico nos municípios de Autazes, Eirunepé e São Gabriel da Cachoeira (2000 a 2005).

In 2005, Amazonas State, Brazil, showed hyperendemic leprosy detection coefficients and prevalence with medium endemicity. Although this State has the largest indigenous population in Brazil, there are no data on the leprosy profile in these groups. This study aimed to describe and analyze the epidemiological characteristics of leprosy case reporting in the municipalities (counties) of Autazes, Eirunepé, and São Gabriel da Cachoeira, comparing indigenous and non-indigenous findings according to target variables. A total of 386 cases reported to SINAN from 2000 to 2005 were analyzed. Mean detection rates were 3.55, 14.94, and 2.13/10,000 (among non-indigenous) and 10.95, 1.93, and 0.78/10,000 (among indigenous peoples) in Autazes, Eirunepé, and São Gabriel da Cachoeira, respectively. Paucibacillary cases predominated among both indigenous and non-indigenous populations; however, dimorphous cases represented one-third of notifications. Despite coverage limitations and underreporting, the findings suggest that leprosy is a major public health problem for indigenous populations in Amazonas State. Classification according to race/ethnicity has been a useful tool for solving health inequalities.

BCG revaccination does not protect against leprosy in the Brazilian Amazon: a cluster randomised trial.

BACKGROUND: Although BCG has been found to impart protection against leprosy in many populations, the utility of repeat or booster BCG vaccinations is still unclear. When a policy of giving a second BCG dose to school children in Brazil was introduced, a trial was conducted to assess its impact against tuberculosis, and a leprosy component was then undertaken in parallel. OBJECTIVE: to estimate the protection against leprosy imparted by a second dose of BCG given to schoolchildren. METHODS AND FINDINGS: This is a cluster randomised community trial, with 6 years and 8 months of follow-up. STUDY SITE: City of Manaus, Amazon region, a leprosy-endemic area in Brazil. PARTICIPANTS: 99,770 school children with neonatal BCG (aged 7-14 years at baseline), of whom 42,662 were in the intervention arm (revaccination). INTERVENTION: BCG given by intradermal injection. MAIN OUTCOME: Leprosy (all clinical forms). RESULTS: The incidence rate ratio of leprosy in the intervention over the control arm within the follow-up, in schoolchildren with neonatal BCG, controlled for potential confounders and adjusted for clustering, was 0.99 (95% confidence interval: 0.68 to 1.45). CONCLUSIONS/SIGNIFICANCE: There was no evidence of protection conferred by the second dose of BCG vaccination in school children against leprosy during the trial follow-up. These results point to a need to consider the effectiveness of the current policy of BCG vaccination of contacts of leprosy cases in Brazilian Amazon region.

Hanseníase em populações indígenas do Amazonas, Brasil: um estudo epidemiológico nos municípios de Autazes, Eirunepé e São Gabriel da Cachoeira (2000 a 2005) / Leprosy in indigenous populations of Amazonas State, Brazil: an epidemiological study in the counties of Autazes, Eirunepé and São Gabriel da Cachoeira (2000 to 2005)

In 2005, Amazonas State, Brazil, showed hyperendemic leprosy detection coefficients and prevalence with medium endemicity. Although this State has the largest indigenous populationin Brazil, there are no data on the leprosy profile in these groups. This study aimed to describe and analyze the epidemiological characteristics of leprosy case reporting in the municipalities (counties) of Autazes, Eirunepé, and São Gabriel da Cachoeira, comparing indigenous and nonindigenousfindings according to target variables.A total of 386 cases reported to SINAN from 2000 to 2005 were analyzed. Mean detection rates were 3.55, 14.94, and 2.13/10,000 (among nonindigenous) and 10.95, 1.93, and 0.78/10,000 (among indigenous peoples) in Autazes, Eirunepé, and São Gabriel da Cachoeira, respectively. Paucibacillary cases predominated among both indigenous and non-indigenous populations; however, dimorphous cases represented one-thirdof notifications. Despite coverage limitations and underreporting, the findings suggest that leprosy is a major public health problem for indigenouspopulations in Amazonas State. Classification according to race/ethnicity has been a useful tool for solving health inequalities.

O Estado do Amazonas, Brasil, apresentou, em 2005, coeficientes hiperendêmicos de detecção de hanseníase e prevalência de média endemicidade. O estado detém a maior população indígena no país, mas inexistem informações sobre o perfil da hanseníase nesses grupos. O estudo objetivou a descrição e análise das características epidemiológicas das notificações de hanseníase nos municípios de Autazes, Eirunepé e São Gabriel da Cachoeira, comparando achados entre indígenas e não indígenas, segundo variáveis de interesse. Foram analisados os casos notificados no SINAN, no período de 2000 a 2005. Do total de 386 casos notificados, verificaram- se coeficientes médios de detecção de 3,55, 14,94 e 2,13/10 mil (entre os não indígenas) e de 10,95, 1,93 e 0,78/10 mil (para os indígenas), para Autazes,Eirunepé e São Gabriel da Cachoeira, respectivamente. Houve predomínio de casos paucibacilares em indígenas e em não indígenas, no entanto, a forma dimorfa representou 1/3 das notificações. Apesar das limitaçõesde cobertura e do sub-registro, os achados sugerem que a hanseníase representa importante problema de saúde pública para os indígenas no Amazonas. A classificação segundo “raça/etnicidade” se constituiu em ferramenta útil para elucidar desigualdades em saúde.

Design of the leprosy component of the Brazilian BCG revaccination trial for assessing BCG effectiveness against leprosy in school children.

BACKGROUND: BCG vaccination confers protection against leprosy, and vaccination among household contacts has been recommended in Brazil. Nevertheless, vaccination of the entire community against leprosy is not advocated as leprosy has low incidence in most populations. Despite that, in Brazil, BCG vaccination is recommended among school children to prevent tuberculosis and this large scale vaccination may also affect the occurrence of leprosy, which led to investigations of its impact on leprosy in endemic areas of Brazil. OBJECTIVES: To estimate the effectiveness against leprosy of a dose of BCG vaccine given to school children in a population with a high coverage of neonatal BCG. Long term objectives are to compare the impact of vaccination among schoolchildren with the existing recommendation to vaccinate household contacts of leprosy. STUDY DESIGN: Cluster randomized controlled field trial with no placebo. STUDY POPULATION: Children aged 7 to 14 years attending state schools with high coverage of neonatal BCG. METHODS: 286 state schools in the city of Manaus, Brazil, were randomized to receive BCG or not. Identifying information was collected for 152,438 school children, of whom 72,980 are in intervention schools. BCG vaccination was given intradermically to children in schools allocated to vaccination. Follow-up relies on ascertainement of cases diagnosed at the health services and notified to the reference center for leprosy.