Lessons from the historical dynamics of environmental law enforcement in the Brazilian Amazon.

Here, we analyze critical changes in environmental law enforcement in the Brazilian Amazon between 2000 and 2020. Based on a dataset of law enforcement indicators, we discuss how these changes explain recent Amazon deforestation dynamics. Our analysis also covers changes in the legal prosecution process and documents a militarization of enforcement between 2018 and 2022. From 2004 to 2018, 43.6 thousand land-use embargoes and 84.3 thousand fines were issued, targeting 3.3 million ha of land, and totaling USD 9.3 billion in penalties. Nevertheless, enforcement relaxed and became spatially more limited, signaling an increasing lack of commitment by the State to enforcing the law. The number of embargoes and asset confiscations dropped by 59% and 55% in 2019 and 2020, respectively. These changes were accompanied by a marked increase in enforcement expenditure, suggesting a massive efficiency loss. More importantly, the creation of so-called conciliation hearings and the centralization of legal processes in 2019 reduced the number of actual judgments and fines collected by 85% and decreased the ratio between lawsuits resulting in paid fines over filed ones from 17 to 5%. As Brazil gears up to crack-down on illegal deforestation once again, our assessment suggests urgent entry points for policy action.

Leishmania infantum infection rate in dogs housed in open-admission shelters is higher than of domiciled dogs in an endemic area of canine visceral leishmaniasis. Epidemiological implications.

Canine visceral leishmaniasis (CVL) is caused by Leishmania infantum and is endemic in many areas of southeastern Brazil. We have hypothesized that the prevalence of infection by L. infantum in dogs housed in open-admission animal shelters is beyond the range of 3.4 - 9.6% reported among dogs domiciled in similar CVL-endemic areas. Hence, this study aimed to determine the rate of L. infantum infection among dogs maintained in shelters and to investigate the epidemiology of CVL in such environments by analyzing hematological and biochemical parameters. A total of 627 dogs from 17 different shelters across the State of Minas Gerais were screened using the Dual-Path Platform test and enzyme-linked immunosorbent assay and 211 (33.6%) were found to be seropositive in both tests. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was performed on skin, bone marrow and lymphoid tissues of 118 seropositive dogs with inconclusive CVL clinical diagnosis and, of these, 78 (66.1%) were PCR+ for L. infantum and 7 (5.9%) were PCR+ for L. amazonensis. One dog presented a PCR-RFLP profile that was consistent with co-infection by both parasites. Leishmania amazonensis DNA was detected in skin samples of six single-infected dogs and this constitutes a novel finding. Dogs infected only with L. amazonensis were less debilitated than those infected by L. infantum, which showed typical clinical manifestations of CVL. The co-infected dog showed only mild clinical signs. The results presented herein not only support our original hypothesis but also suggest that dogs are potential reservoirs of L. amazonensis. Public health authorities should acknowledge their responsibility towards animals in collective shelters, recognize that they are potential foci of zoonotic diseases, and establish proper functioning directives to minimize transmission to humans and to other dogs.

Evidências científicas sobre hérnia de disco na pessoa idosa / Scientific evidence on disk hernia in elderly person / Evidencia científica sobre el disco hernia en persona mayor

Objetivo: identificar as produções científicas sobre hérnia de disco na pessoa idosa. Método: protocolo de scoping review segundo o método do Joanna Briggs Institute com artigos das bases de dados CINAHL, Scopus, Cochrane, Web of Science, MEDLINE e ScienceDirect. Analisaram-se 10 artigos cujos critérios de inclusão foram investigações primárias e/ou secundárias, qualitativas e quantitativas escritos em inglês, português e espanhol, formando 3 categorias temáticas. O recorte do estudo foi delimitado a partir da observação na prática profissional dos pesquisadores sobre o aumento de casos de idosos com hérnia de disco. Resultados: a cirurgia representa constante indicação médica para o tratamento da hérnia de disco. Conclusão:a lombalgia decorrente da hérnia de disco é uma frequente causa de incapacidade nos adultos e idosos com protocolos cirúrgicos constantes, principalmente quando há compressão neural. No entanto, estudos mostram a possibilidade de regressão quando os pacientes são submetidos aos tratamentos conservadores

Objective: to identify scientific productions about herniated discs in the elderly. Method: scoping review protocol according to the Joanna Briggs Institute method with articles from the CINAHL, Scopus, Cochrane, Web of Science, MEDLINE and ScienceDirect databases. Were analyzed 10 articles whose inclusion criteria were primary and / or secondary, qualitative and quantitative investigations written in English, Portuguese and Spanish, forming 3 thematic categories. The study was delimited based on the observation in the professional practice of researchers about the increase in cases of elderly people with herniated discs. Results:surgery is a constant medical indication for the treatment of herniated discs. Conclusion: low back pain due to herniated discs is a frequent cause of disability in adults and the elderly with constant surgical protocols, especially when there is neural compression. However, studies show the possibility of regression when patients are submitted to conservative treatments

Objetivo: identificar producciones científicas sobre hernias discales en ancianos. Método: protocolo de revisión del alcance según el método del Instituto Joanna Briggs con artículos de las bases de datos CINAHL, Scopus, Cochrane, Web of Science, MEDLINE y ScienceDirect. Se analizaron 10 artículos cuyos criterios de inclusión fueron investigaciones primarias y / o secundarias, cualitativas y cuantitativas escritas en inglés, portugués y español, formando 3 categorías temáticas. El estudio se delimitó en función de la observación en la práctica profesional de los investigadores sobre el aumento de casos de personas mayores con discos herniados. Resultados: la cirugía representa una indicación médica constante para el tratamiento de discos herniados. Conclusión:el dolor lumbar debido a discos herniados es una causa frecuente de discapacidad en adultos y ancianos con protocolos quirúrgicos constantes, especialmente cuando hay compresión neural. Sin embargo, los estudios muestran la posibilidad de regresión cuando los pacientes son sometidos a tratamientos conservadores

Detection of Metalloproteases and Cysteine Proteases RNA Transcripts of Leishmania (Leishmania) infantum in Ear Edge Skin of Naturally Infected Dogs.

Leishmania spp. proteases have been proposed as virulence factors contributing to adaptive success these parasites to the mammalian hosts. Since these enzymes are poorly studied in naturally infected dogs, this work aims to show the differences in metalloprotease and cysteine proteases gene expression in ear edge skin of dogs naturally infected by Leishmania (Leishmania) infantum. A cohort of dogs (n = 20) naturally infected by L. (L.) infantum was clinically classified as asymptomatic, oligosymptomatic, and polysymptomatic and the parasite load range estimated. The analysis of proteases expression by RT-PCR in the ear edge skin was also assessed, suggesting more transcripts of proteases in cDNA samples from polysymptomatic dogs than oligosymptomatic and asymptomatic ones. Metalloprotease RT-PCR assays yielded products (202 bp) in all assessed cDNA dog samples. In contrast, cysteine proteases transcripts (227 bp) had shown to be better detected in cDNA samples of polysymptomatic dogs, compared with cDNA samples from asymptomatic and oligosymptomatic dogs. Predictive in silico assays suggested that secondary structures of metalloproteasee mRNAs can be more stable than cysteine proteases at the skin temperature of dogs. Evidence is presented that during natural infection of dogs by L. (L.) infantum, this parasite produces transcripts of metalloprotease and cysteine protease RNA in the skin from asymptomatic, oligosymptomatic, and polysymptomatic dogs.

Reactivity of sera from dogs living in a leishmaniasis-endemic area to the COOH-terminal region of cysteine proteinase B

ABSTRACT Cysteine proteinases are well-known virulence factors of Leishmania spp. with demonstrated actions in both experimental mouse infection and human infection. However, studies on these enzymes in canine leishmaniasis are scarce. Here, we show, for the first time, the reactivity of sera from dogs living in an endemic area to a recombinant protein from the COOH-terminal region of cysteine B protease. In this work, enzyme-linked immunosorbent assays were performed using a 14 kDa rcyspep protein obtained through a pET28-a expression system in Escherichia coli. First, 96-well plates were coated with rcyspep (500 ng/well) and incubated with sera from dogs (1:100). Subsequently, IgG antibody detection was performed using rabbit anti-dog IgG antibodies conjugated with peroxidase. Sera from dogs (n = 114), including suspect (n = 30) and positive (n = 50) dogs from a leishmaniasis-endemic area and dogs from a nonendemic area, (n = 34), negative for leishmaniasis, were assessed. The results showed that sera from the suspect (42%) and positive (68%) groups responded differently to the antigen titers tested above the cut-off (Optical Density = 0.166). This finding suggests that the immune response detected against cyspep may be related to clinical disorders present in these animals. Collectively, the data gathered here suggest that cyspep can sensitize the immune systems of dogs from a leishmaniasis-endemic area to elicit a humoral response, an immunological parameter indicating the contribution of this protein in host-parasite interaction.

Reactivity of sera from dogs living in a leishmaniasis-endemic area to the COOH-terminal region of cysteine proteinase B.

Cysteine proteinases are well-known virulence factors of Leishmania spp. with demonstrated actions in both experimental mouse infection and human infection. However, studies on these enzymes in canine leishmaniasis are scarce. Here, we show, for the first time, the reactivity of sera from dogs living in an endemic area to a recombinant protein from the COOH-terminal region of cysteine B protease. In this work, enzyme-linked immunosorbent assays were performed using a 14kDa rcyspep protein obtained through a pET28-a expression system in Escherichia coli. First, 96-well plates were coated with rcyspep (500ng/well) and incubated with sera from dogs (1:100). Subsequently, IgG antibody detection was performed using rabbit anti-dog IgG antibodies conjugated with peroxidase. Sera from dogs (n=114), including suspect (n=30) and positive (n=50) dogs from a leishmaniasis-endemic area and dogs from a nonendemic area, (n=34), negative for leishmaniasis, were assessed. The results showed that sera from the suspect (42%) and positive (68%) groups responded differently to the antigen titers tested above the cut-off (Optical Density=0.166). This finding suggests that the immune response detected against cyspep may be related to clinical disorders present in these animals. Collectively, the data gathered here suggest that cyspep can sensitize the immune systems of dogs from a leishmaniasis-endemic area to elicit a humoral response, an immunological parameter indicating the contribution of this protein in host-parasite interaction.

Drogas: uma análise semântica dos estudos brasileiros / Drugs: a semantic analysis of brazilian studies / Drogas: un análisis semántico de los estudios brasileños

OBJETIVO: realizar uma análise semântica dos principais elementos discursivos presentes em publicações nacionais sobre drogas ilícitas. MÉTODO: foram utilizados 336 trechos de resumos retirados de 58 artigos publicados em revistas brasileiras entre 2004 e 2014. Quando submetidos ao programa gratuito Iramuteq®, obteve-se a frequência e a classe das palavras, que revelaram os principais elementos discursivos sobre a temática "drogas". RESULTADOS: os dados se estruturaram em cinco classes divididas em dois grupos, que revelam a frequência do corpus de palavras analisadas: Dados e Usuário, de modo que o grupo "Dados" foi composto pelas classes: a) Epidemiologia (17,5%) e b) Método de pesquisa (31,4%). Já o grupo "Usuário", pelas classes: c) Atenção em saúde (19,1%); d) Consequências do abuso (16,1%); e) Aspectos sociais (15,9%). CONCLUSÃO: os discursos presentes nas perspectivas dos autores apontam a preferência pelas pesquisas investigativas, pela utilização do método quantitativo e as pesquisas relacionaram a temática das drogas a aspectos sócio-políticos e à necessidade de novos planejamentos dos serviços de atenção em saúde.

OBJECTIVE: is to perform a semantic analysis of the discursive elements present in national publications on illicit drugs. METHOD: a total of 336 excerpt extracts from 58 articles published in Brazilian journals between 2004 and 2014 were used. The articles were submitted to the Iramuteq® program and the frequency and class of words were obtained, which revealed the main discursive elements on the theme 'drugs'. RESULTS: the data were structured into five classes divided into two groups, which reveal the frequency of the corpus of analyzed words: Data and User, so that the "Data" group was composed of classes a) Epidemiology (17.5%) and b) Method (31.4%). The "User" group was composed by classes c) Health care (19.1%); d) Consequences of abuse (16.1%) and e) Social aspects (15.9%). CONCLUSION: the results indicated that the present discourses, in authors perspectives, point to the preference for investigative research, using the quantitative method, and the research related the thematic of drugs to socio-political aspects and the need for new plans of health care services.

OBJECTIVO: realizar un análisis semántico de los principales elementos discursivos presentes en publicaciones nacionales sobre drogas ilícitas. MÉTODO: se utilizaron 336 extractos de resúmenes retirados de 58 artículos publicados en revistas brasileñas entre 2004 y 2014. Cuando se sometieron al programa gratuito Iramuteq®, se obtuvo la frecuencia y la clase de las palabras, que revelaron los principales elementos discursivos sobre la temática 'drogas'. RESULTADOS: los datos se estructuraron en cinco clases divididas en dos grupos, que revelan la frecuencia del corpus de palabras analizadas: Datos y Usuario, de modo que el grupo "Datos" fue compuesto por las clases a) Epidemiología (17,5%) yb) Método de investigación (31,4%). El grupo "Usuario", por las clases c) Atención en salud (19,1%); d) Consecuencias del abuso (16,1%); e) Aspectos sociales (15,9%). CONCLUSIÓN: los resultados indicaron que los discursos presentes en las perspectivas de los autores apuntan la preferencia por las investigaciones investigativas, por la utilización del método cuantitativo, y las investigaciones relacionaron la temática de las drogas a aspectos socio-políticos ya la necesidad de nuevos planificamientos de los servicios de atención en salud.

The combination therapy of meglumine antimoniate and oxiranes (epoxy-α-lapachone and epoxymethyl-lawsone) enhance the leishmanicidal effect in mice infected by Leishmania (Leishmania) amazonensis.

Current treatment of cutaneous leishmaniasis includes pentavalent antimonials as first-line drugs, but this therapy has shown severe adverse effects. An alternative to minimize this issue is based on combination therapy scheme with other drugs. In this study we analyzed the potential of the association of meglumine antimoniate (MA) with the oxiranes epoxy-α-lapachone (LAP) or epoxymethyl-lawsone (LAW). Results demonstrated that association between these drugs enhanced leishmanicidal activity on Leishmania (Leishmania) amazonensis infection. The compounds were tested in monotherapy or in combinations (3:1; 1:1 and 1:3) and reduced intracellular parasite numbers, measured by the endocytic index, in all tested conditions. The most effective combination regimens were MA/LAP or MA/LAW in 3:1 ratio, which achieved a reduction of 98.3% and 93.6% in the endocytic index, respectively. BALB/c mice challenged with L. (L.) amazonensis showed significant reduction in lesion size and parasite load in both footpad and lymph nodes, after four weeks of treatment. Although, MA, LAP or LAW monotherapy were able to control the evolution of lesions when compared to untreated animals (30%, 40% and 40% of reduction, respectively), the combination of MA/LAP and LAW in 3:1 ratio showed better results reducing 61.7 and 54.4%, respectively. The results indicate that the association of meglumine antimoniate to oxiranes lead to an increment in the antileishmanial activity and represent a promising approach for the cutaneous leishmaniasis treatment.

Understanding serine proteases implications on Leishmania spp lifecycle.

Serine proteases have significant functions over a broad range of relevant biological processes to the Leishmania spp lifecycle. Data gathered here present an update on the Leishmania spp serine proteases and the status of these enzymes as part of the parasite degradome. The serine protease genes (n = 26 to 28) in Leishmania spp, which encode proteins with a wide range of molecular masses (35 kDa-115 kDa), are described along with their degrees of chromosomal and allelic synteny. Amid 17 putative Leishmania spp serine proteases, only ∼18% were experimentally demonstrated, as: signal peptidases that remove the signal peptide from secretory pre-proteins, maturases of other proteins and with metacaspase-like activity. These enzymes include those of clans SB, SC and SF. Classical inhibitors of serine proteases are used as tools for the characterization and investigation of Leishmania spp. Endogenous serine protease inhibitors, which are ecotin-like, can act modulating host actions. However, crude or synthetic based-natural serine protease inhibitors, such as potato tuber extract, Stichodactyla helianthus protease inhibitor I, fukugetin and epoxy-α-lapachone act on parasitic serine proteases and are promising leishmanicidal agents. The functional interrelationship between serine proteases and other Leishmania spp proteins demonstrate essential functions of these enzymes in parasite physiology and therefore their value as targets for leishmaniasis treatment.

Representações sociais da depressão construídas por idosos e suas relações com a capacidade funcional / Social Representations of Depression Built by Elders and their Relation to Functional Capacity

Pesquisas atuais indicam que a cada ano, cerca de 650 mil de pessoas com mais de 60 anos ingressam na sociedade brasileira, sendo a maior parte com doenças crônicas e limitações funcionais. Entre as patologias crônicas, destaca-se a depressão, elevando ainda mais o ciclo de fragilidade e de deterioração progressiva da capacidade funcional. Diante do exposto, o presente estudo tem por objetivos verificar as representações sociais construídas por idosos sobre a depressão na velhice e suas relações com a capacidade funcional, bem como investigar a associação entre os sintomas depressivos e a capacidade funcional. Trata-se, portanto, de um estudo exploratório, de abordagem mista, e de base de dados secundária, que incluiu 234 pessoas com mais de 60 anos, de ambos os sexos, assistidos pelas equipes de saúde da Estratégia Saúde da Família Grotão I, II e III, e a Comunidade Maria de Nazaré, no município de João Pessoa, Paraíba, Brasil. Os dados foram coletados nas respectivas unidades de saúde, através de uma entrevista semiestruturada, composta por três partes, sendo a primeira o Teste de Associação Livre de Palavras (TALP), com o termo indutor ; seguida pela caracterização sociodemográfica; a última parte foi contemplada pela Escala de Depressão Geriátrica (GDS-15) e análise do desempenho funcional, por meio do Índice de Katz e da Escala de Lawton e Brody. Os dados coletados foram processados pelos softwares SPSS versão 21.0 e IRaMuTeQ e, posteriormente interpretados de acordo com o aporte teórico das Representações Sociais. Verificou-se que 38% (n=88) dos idosos participantes apresentaram sintomas sugestivos de depressão, com prevalência entre as mulheres, na faixa etária entre 60 a 69 anos. Foi possível observar que uma significativa parcela da população (43,6%) se identifica como pessoas independentes e sem sintomas de depressão, embora, 13,7% se classificam como sendo independentes pelo índice de Katz e dependentes parcialmente pela escala de Lawton. Cabe destacar que, 9,83% dos participantes que apresentavam sintomas depressivos já demonstravam também perdas funcionais para atividades mais complexas, que requerem habilidades cognitivas e motoras associadas, como se observa nas Atividades Instrumentais de Vida Diária. A análise do corpus textual referente à representações sociais da depressão resultou na formação de quatro classes semânticas, com aproveitamento de 77,68%, apresentadas pela Classificação Hierárquica Descendente (CHD): classe 1 - Sentidos e experiências associadas à depressão; classe 2 - Aspectos relacionados à depressão; classe 3 - Aspectos clínicos da depressão; classe 4 - Dimensões psicoafetivas da depressão. Os idosos considerados funcionalmente dependentes representam a depressão como um processo patológico grave de repercussão psicoafetiva, que necessita de assistência clínica e medicamentosa, mas que o cuidado se torna mais ampliado e efetivo com a presença da família e de amigos. Os dados do presente estudo correlacionam-se entre si, e indicam que a presença dos sintomas depressivos está associada significativamente ao declínio progressivo das Atividades Instrumentais de Vida Diária, assim como, com a perda progressiva do desempenho cognitivo, sugerindo atualizações aos profissionais de saúde através das práticas e dos norteamentos direcionados aos idosos, com e sem sintomas depressivos. (AU)

Current researches indicate that, every year, around 650,000 people over 60 are entering Brazilian society, most of them with chronic diseases and functional limitations. Among the chronic pathologies, depression stands out, further increasing the cycle of fragility and progressive deterioration of functional capacity. Therefore, the present study aims to verify the social representations built by the elders on depression in old age and its relation with functional capacity, as well as to investigate the association between depressive symptoms and functional capacity. This is an exploratory study, with mixed approach, and secondary database, which included 234 people over 60 years old, of both sexes, assisted by the health teams of the Family Health Strategy Grotão I, II and III, and the Maria de Nazaré Community, in the municipality of João Pessoa, Paraíba, Brazil. The data were collected in the respective health units, through a semi-structured interview, composed by three parts, the first being the Free Word Association Test (TALP), with the inductor term ; followed by sociodemographic characterization; the last part was contemplated by the Geriatric Depression Scale (GDS-15) and functional performance analysis, using the Katz Index and the Lawton and Brody Scale. The data collected were processed by the software SPSS version 21.0 and IRaMuTeQ and later interpreted according to the theoretical contribution of the Social Representations. The results show that 38% (n=88) of the elderly participants presented symptoms suggestive of depression, prevalent among women, in the age group between 60 and 69 years. Moreover, a significant portion of the population (43.6%) were identified as independent persons with no symptoms of depression, although 13.7% self-classified as independent by the Katz index and partially dependent on the Lawton scale. Furthermore, 9.83% of participants with depressive symptoms already showed functional losses for more complex activities, which require associated cognitive and motor skills, as observed in the Instrumental Activities of Daily Living. The analysis of the textual corpus referring to the social representations of depression resulted in the formation of four semantic classes, with use of 77.68%, presented by the Descending Hierarchical Classification (CHD): class 1 - Senses and experiences associated with depression; Class 2 - Aspects related to depression; Class 3 - Clinical aspects of depression; Class 4 - Psychoaffective dimensions of depression. The elders considered functionally dependent represent depression as a serious pathological process of psychoaffective repercussion, which requires clinical and medication assistance, but that care becomes more extended and effective with the presence of family and friends. Data from the present study correlate with each other, and indicate that the presence of depressive symptoms is significantly associated with the progressive decline of Instrumental Activities of Daily Living, as well as with the progressive loss of cognitive performance, suggesting updates to health professionals through practices and orientations directed to the elders, with and without depressive symptoms. (AU)

A defective TLR4 signaling for IFN-ß expression is responsible for the innately lower ability of BALB/c macrophages to produce NO in response to LPS as compared to C57BL/6.

C57BL/6 mice macrophages innately produce higher levels of NO than BALB/c cells when stimulated with LPS. Here, we investigated the molecular events that account for this intrinsic differential production of NO. We found that the lower production of NO in BALB/c is not due to a subtraction of L-arginine by arginase, and correlates with a lower iNOS accumulation, which is independent of its degradation rate. Instead, the lower accumulation of iNOS is due to the lower levels of iNOS mRNA, previously shown to be also independent of its stability, suggesting that iNOS transcription is less efficient in BALB/c than in C57BL/6 macrophages. Activation of NFκB is more efficient in BALB/c, thus not correlating with iNOS expression. Conversely, activation of STAT-1 does correlate with iNOS expression, being more prominent in C57BL/6 than in BALB/c macrophages. IFN-ß and IL-10 are more highly expressed in C57BL/6 than in BALB/c macrophages, and the opposite is true for TNF-α. Whereas IL-10 and TNF-α do not seem to participate in their differential production of NO, IFN-ß has a determinant role since 1) anti-IFN-ß neutralizing antibodies abolish STAT-1 activation reducing NO production in C57BL/6 macrophages to levels as low as in BALB/c cells and 2) exogenous rIFN-ß confers to LPS-stimulated BALB/c macrophages the ability to phosphorylate STAT-1 and to produce NO as efficiently as C57BL/6 cells. We demonstrate, for the first time, that BALB/c macrophages are innately lower NO producers than C57BL/6 cells because they are defective in the TLR-4-induced IFN-ß-mediated STAT-1 activation pathway.

Performance of an indigenous ß-mercaptoethanol-modified antigen in comparison with a commercial reference in direct agglutination test for detection of canine visceral leishmaniasis.

We compared the performance of a locally produced ß-mercaptoethanol-modified promastigote antigen (ß-ME-Ag) of an indigenous Leishmania infantum strain against that of a trypsinized Leishmania donovani reference (REF-Ag) in the direct agglutination test (DAT) for detection of canine visceral leishmaniasis (CVL). One hundred and fifty-one serum samples collected from dogs belonging to four groups with different conditions were included. At a DAT titre of 1 : 320, statistically determined as optimal cut-off value for ß-ME-Ag, and 1 : 160 for REF-Ag, a sensitivity and a specificity of 100 % were estimated for ß-ME-Ag in comparison with 96.6 % and 100 %, respectively, for REF-Ag. Overall, levels of agglutination titres recorded for the two antigens were highly concordant (Cohen's κ = 0.879) in both the CVL and non-CVL groups. Based on current results, and ease experienced in processing the antigen and reading the test outcome, we recommend incorporation of ß-ME-Ag in DAT for confirmation or exclusion of suspected CVL in dogs.

A ß-mercaptoethanol-modified enzyme-linked immunosorbent assay for diagnosis of canine visceral leishmaniasis.

Two immunoglobulin G enzyme-linked immunosorbent assay (ELISA) versions using whole promastigotes of Leishmania infantum (syn. Leishmania chagasi) treated either with ß-mercaptoethanol (ß-ME-ELISA) or trypsin (TRYP-ELISA) as antigens were developed for the diagnosis of canine visceral leishmaniasis (CVL). By comparison with the direct agglutination test (DAT; 100%, 31/31; 95% confidence interval [CI]: 86.3-100%), slightly lower sensitivity was demonstrated for the newly developed ß-ME-ELISA (93.5%, 29/31; 95% CI: 77.2-98.9%). Sensitivity was higher for ß-ME-ELISA compared with TRYP-ELISA (87.1%, 27/31; 95% CI: 69.2-95.8%) in serum samples from dogs with CVL. When tested with sera from 37 healthy dogs and from 45 dogs with clinical conditions other than CVL, a specificity of 97.6% (80/82; 95% CI: 90.1-99.6%) was estimated for ß-ME-ELISA as compared to 100% (82/82; 95% CI: 94.4-100%) and 95.1% (78/82; 95% CI: 87.3-98.4%) for DAT and TRYP-ELISA, respectively. Observed agreement was 94.0% (95% CI: 88.7-97.1%) between DAT and ß-ME-ELISA (κ = 0.879; 95% CI: 0.803-0.956) and 87.4% (95% CI: 80.8-92.1%) between DAT and TRYP-ELISA (κ = 0.743; 95% CI: 0.636-0.851). Current results advocate application of the new ß-ME-ELISA for diagnosis of CVL at the laboratory level and confirmation of results obtained with the DAT in field studies.