RESUMEN
The aim of the study was to evaluate the association between salivary anti-Porphyromonas gingivalis IgA antibodies and the leprosy reaction. The levels of salivary anti - P. gingivalis IgA antibodies, together with salivary flow and pH were measured in individuals diagnosed with leprosy and associated with the development of the leprosy reaction. Saliva was collected from 202 individuals diagnosed with leprosy at a reference leprosy treatment center, 106 cases with the leprosy reaction and 96 controls without the leprosy reaction. Anti - P. gingivalis IgA was evaluated by indirect immunoenzyme assay. Non-conditional logistic regression analysis was employed to estimate the association between antibody levels and the leprosy reaction. There was a positive statistically significant association between the levels of anti - P. gingivalis IgA and the presence of the leprosy reaction, controlling for confounders: age, sex, level of education and alcoholic beverage consumption: ORajusted: 2.55; IC 95%: 1.34-4.87. Individuals with leprosy who had high levels of salivary anti - P. gingivalis IgA had approximately twice as many chances of developing the leprosy reaction. The findings suggest a possible relationship between salivary anti - P. gingivalis IgA antibodies and the leprosy reaction.
RESUMEN
Caseous lymphadenitis (CLA) is a chronic disease that affects small ruminants and causes economic losses in the associated breeding system. The causative agent of CLA is Corynebacterium pseudotuberculosis, a Gram-positive bacterium that exhibits tropism for external and internal lymph nodes and induces abscess formation in the host. Bacterial communities often produce a biofilm matrix that serves various functions, including protection against hostile environmental conditions, antibiotics, and the host immune response. Although biofilm formation has been reported for C. pseudotuberculosis, not all strains demonstrate this property in culture. In this work, we report the first comparative proteomic analysis of one biofilm-forming (CAPJ4) and one biofilm-non-forming strain (CAP3W) of C. pseudotuberculosis isolated from goats. Bacterial whole cell protein extracts were obtained for mass spectrometry analyses. Using LC-MS/MS, our studies reveal three and four proteins exclusively found in the CAPJ4 and CAP3W proteome, respectively. In addition, label-free quantitative analysis identified 40 proteins showing at-least 2-fold higher values in CAPJ4 compared CAP3W proteome Notably, CAPJ4 differentially synthesized the penicillin-binding protein, which participates in the formation of peptidoglycans. CAPJ4 also exhibited upregulation of N-acetylmuramoyl-L-alanine amidase and galactose-1-phosphate uridylyltransferase, which are involved in biofilm formation and exopolysaccharide biosynthesis. Here, we demonstrate that biofilm formation in C. pseudotuberculosis is likely associated with specific proteins, some of which were previously shown to be associated with virulence and biofilm formation in other organisms. Our findings may drive studies related to the bacterial mechanisms involved in the biofilm formation, in addition to providing targets for the treatment of CLA.
RESUMEN
Caseous lymphadenitis (CL) is a chronic infectious disease that affects sheep and goats. Many serological tests have been developed to detect the disease; one of the most widely used is the enzyme-linked immunosorbent assay (ELISA), due to its advantages, which include acceptable cost-effectiveness, applicability, sensitivity and specificity. ELISA formulations using recombinant proteins can exhibit significant sensitivity and specificity when using a single purified antigen. DTxR, Trx, TrxR, LexA, SodC, SpaC, NanH, and PknG recombinant proteins can be considered target proteins for ELISA development due to its extracellular or on the cell surface location, which allows a better recognition by the immune system. Therefore, the objectives of this study were to evaluate the antigenic reactivity of Corynebacterium pseudotuberculosis recombinant proteins in goat and sheep serum. Of eight proteins evaluated, rSodC was selected for validation assays with small ruminant serum samples from the semiarid region of the state of Bahia, Brazil. Validation assays with goat serum samples showed that ELISA-rSodC presented sensitivity and specificity of 96% and 94%, respectively. Validation assays with sheep serum showed that ELISA-rSodC exhibited sensitivity and specificity of 95% and 98%, respectively. Analysis of 756 field serum samples showed that rSodC identified 95 positive samples (23%) in goats and 75 positive samples (21%) in sheep. The ELISA with recombinant SodC protein developed in this study discriminated positive and negative serum samples with high levels of sensitivity and specificity. This formulation is promising for epidemiological surveys and CL control programs.Trial registration AEC No 4958051018. 12/18/2018, retrospectively registered.
RESUMEN
Objetivou-se estudar a cinética da proteína total, fibrinogênio e ceruloplasmina durante os primeiros cinco meses de vida, em cordeiros saudáveis da raça Santa Inês, no município de São Gonçalo dos Campos, Bahia, Brasil. Amostras de sangue foram colhidas de 22 animais, ao longo de onze momentos: logo após o parto (T0), 12 (T1), 24 (T2), 48 horas (T3), sete (T4), 15 (T5), 30 (T6), 60 (T7), 90 (T8), 120 (T9) e 150 dias de vida (T10). A proteína total e o fibrinogênio plasmáticos foram analisados por meio de refratômetro clínico e pela técnica de desnaturação pelo calor, respectivamente, enquanto que a determinação da ceruloplasmina sérica se baseou em sua atividade oxidásica. Para análise estatística utilizou-se o programa SPSS versão 18; os dados com distribuição não paramétrica foram submetidos ao teste de Friedman para avaliar o efeito do tempo, enquanto que as comparações múltiplas pelo teste de Wilcoxon permitiram a identificação das diferenças entre os momentos, adotando-se nível de significância de 5% (p<0,05). A proteína total apresentou o menor valor no T0 diferindo estatisticamente dos demais tempos, com pico às 12 horas (T1), porém estabilizando-se até o final do experimento. O fibrinogênio não apresentou diferença estatística entre os tempos. De T1 (12h) a T3 (48h) constatou-se baixos valores de ceruloplasmina, muito embora às 24 horas (T2) tenha diferido estatisticamente (p<0,05), em relação ao T0. A partir do sétimo dia (T4) a concentração desta proteína aumentou significativamente, atingindo pico nos tempos T8 (90 dias) e T9 (120 dias). Foi possível estabelecer a cinética das proteínas estudadas, identificar os principais momentos com alterações e sugerir os fatores associados com as mudanças observadas.
The objective was to study the kinetics of the total protein, fibrinogen, and ceruloplasmin during the first five months of life in healthy lambs Santa Inês, in São Gonçalo dos Campos, Bahia, Brazil. Blood samples were collected from 22 animals, over eleven times: immediately after birth (T0), 12 (T1), 24 (T2), 48 hours (T3), seven (T4), 15 (T5), 30 (T6), 60 (T7), 90 (T8) 120 (T9), and 150 days of age (T10). The total protein and plasma fibrinogen were analyzed by means of a clinical refractometer and the heat denaturation technique, respectively, while the determination of serum ceruloplasmin was based on its oxidase activity. For statistical analysis, the SPSS version 18 program was used; the non-parametric data were submitted to the Friedman test to evaluate the effect of time, whereas the multiple comparisons by the Wilcoxon test allowed the identification of the differences between the moments, adopting a significance level of 5% (P < 0.05). The total protein presented the lowest value at T0 differing statistically from the other times, with a peak at 12 hours (T1), but stabilizing until the end of the experiment. Fibrinogen is not able to differentiate between the times. Between T1 (12h) and T3 (48h), low values of ceruloplasmin were observed, although at 24 hours (T2) it differed statistically (p <0.05) in relation to T0. On the seventh day (T4) the concentration of this protein increased significantly, reaching a peak at T8 (90 days) and T9 (120 days). It was possible to establish the kinetics of the proteins studied, to identify the main moments with alterations and to suggest the factors associated with the observed changes.