Inheritance of genomic regions and genes associated with number of oocytes and embryos in Gir cattle through daughter design.

Over the past decades, daughter designs, including genotyped sires and their genotyped daughters, have been used as an approach to identify QTL related to economic traits. The aim of this study was to identify genomic regions inherited by Gir sire families and genes associated with number of viable oocytes (VO), total number of oocytes (TO), and number of embryos (EMBR) based on a daughter design approach. In total, 15 Gir sire families were selected. The number of daughters per family ranged from 26 to 395, which were genotyped with different SNP panels and imputed to the Illumina BovineHD BeadChip (777K) and had phenotypes for oocyte and embryo production. Daughters had phenotypic data for VO, TO, and EMBR. The search for QTL was performed through GWAS based on GBLUP. The QTL were found for each trait among and within families based on the top 10 genomic windows with the greatest genetic variance. For EMBR, genomic windows identified among families were located on BTA4, BTA5, BTA6, BTA7, BTA8, BTA13, BTA16, and BTA17, and they were most frequent on BTA7 within families. For VO, genomic windows were located on BTA2, BTA4, BTA5, BTA7, BTA17, BTA21, BTA22, BTA23, and BTA27 among families, being most frequent on BTA8 within families. For TO, the top 10 genomic windows were identified on BTA2, BTA4, BTA5, BTA7, BTA17, BTA21, BTA22, BTA26, and BTA27, being most frequent on BTA7 and BTA8 within families. Considering all results, the greatest number of genomic windows was found on BTA7, where the VCAN, XRCC4, TRNAC-ACA, HAPLN1, and EDIL3 genes were identified in the common regions. In conclusion, 15 Gir sire families with 26 to 395 daughters per family with phenotypes for oocyte and embryo production helped to identify the inheritance of several genomic regions, especially on BTA7, where the EDIL3, HAPLN1, and VCAN candidate genes were associated with number of oocytes and embryos in Gir cattle families.

Structure-activity relationships and mechanism of action of tetragomycin derivatives as inhibitors of Staphylococcus aureus staphyloxanthin biosynthesis.

Despite the main strategy to overcome bacterial resistance has focused on the development of more potent antimicrobial agents, the evolutionary pressure caused by such drugs makes this strategy limited. Molecules that interfere with virulence factors appear as a promising alternative though, as they cause reduced selective pressure. As a matter of fact, staphyloxanthin biosynthesis inhibition (STXBI) has been pursued as promising strategy to reduce S. aureus virulence. Herein, we report the inhibitory profile of 27 tetrangomycin derivatives over staphyloxanthin production. The experimental result showed that naphthoquinone dehydro-α-lapachone (25 - EC50 = 57.29 ± 1.15 µM) and 2-Isopropylnaphtho[2,3-b]furan-4,9-dione (26 EC50 = 82.10 ± 1.09 µM) are the most potent compounds and suggest that hydrogen acceptor groups and lipophilic moieties decorating the naphthoquinone ring are crucial for STXBI. In addition, we present an in situ analysis, through RAMAN spectroscopy, that is inexpensive and might be employed to probe the mechanism of action of staphyloxanthin biosynthesis inhibitors. Therefore, our molecular simplification strategies afforded promising lead compounds for the development of drugs that modulate S. aureus staphyloxanthin biosynthesis.

Different methods of assessing udder temperature through thermography and their relation with rectal temperature.

Infrared thermography of the lateral side portion of the udder was taken from 38 lactating cows in a 6-day experiment. Thermograms were analyzed using specific software with the use of eight different methods. The experiment was aimed at correlating the different methods with each other and with rectal temperature (RT), in order to suggest a method to assess udder side temperature, and also at creating regression equations to enable RT calculation through the use of thermographic data. All methods of analyzing thermograms were highly correlated; correlations between thermograms and RT were significant (P < 0.01) and above 0.840. The highest correlation was between RT and the maximum temperature obtained using a horizontal rectangle placed within the lateral (side) portion of the udder (maximum temperature in a horizontal rectangle (MHR), 0.897). With the exception of the average temperature of a horizontal rectangle, linear regression coefficients were significant (P < 0.05) and coefficients of determination were higher than 75.51%. We suggest the use of MHR to evaluate udder side temperature. The ease of accessing the side of the udder, the welfare advantages of non-invasive observations, and the high correlation with RT suggest the use of thermograms in the lateral portion of the udder to assess animals' temperature.

Eating behaviour patterns are associated with excessive weight gain after liver transplantation.

BACKGROUND: Weight gain and obesity are important issues in liver transplant (LTx) recipients. Although dietary habits are probably related to excessive weight gain after LTx, some studies have failed to demonstrate these associations. The present study aimed to assess eating behaviour and verify its association with weight gain and excessive weight among LTx recipients. METHODS: The investigation comprised a cross-sectional study. Post-LTx patients, aged >18 years were evaluated about their eating behaviour [Three Factor Eating Questionnaire-R21 (TFEQ-R21)]. The scores of the TFEQ-R21 [uncontrolled eating (UE), cognitive restraint (CR) and emotional eating (EE)] were determined. Excessive weight was classified as body mass index ≥25 kg/m² and obesity ≥30 kg/m². RESULTS: In total, 270 patients (age 58.0 years; range 20.0-77.0 years; 64.1% men) were assessed. Average weight gain was 8.0 kg (range -16.0 to 41.0 kg). Of the patients, 64.1% (n = 173) had excessive weight and 23.3% (n = 63) were obese. Post-LTx weight gain was significantly correlated with UE (r = 0.311, P < 0.001) and EE (r = 0.287, P < 0.001). Patients with excessive weight had significantly higher scores than others: CR (61.0; range 0-122.0 versus 44.0; range 0-116.0) (P = 0.003) and EE (11.0; range 0-100.0 versus 0.0; range 0-100.0) (P = 0.030). Patients with obesity had higher scores than non-obese UE (22.0; range 0-77.0 versus 14.0; range 0-85.0; P = 0.028), CR (55.0; range 11.0-116.0 versus 50.0; range 0-122.0) (P = 0.017) and EE (16.0; range 0-100.0 versus 5.0; range 0-100.0) individuals (P < 0.001). The greatest quartiles of weight gain had higher scores on the eating behaviours of UE and EE, especially those with weight gain ≥14 kg. UE was associated with weight gain. CR was associated with being overweight. EE was associated with obesity. CONCLUSIONS: Uncontrolled eating is associated with weight gain, CR was associated with excessive weight and EE was associated with obesity after LTx.

RNA sequencing differential gene expression analysis of isolated perfused bovine udders experimentally inoculated with Streptococcus agalactiae.

The aim of this study was to elucidate the differential gene expression in the RNA sequencing transcriptome of isolated perfused udders collected from 4 slaughtered Holstein × Zebu crossbred dairy cows experimentally inoculated with Streptococcus agalactiae. We studied 3 different statistical tools (edgeR, baySeq, and Cuffdiff 2). In summary, 2 quarters of each udder were experimentally inoculated with Strep. agalactiae and the other 2 were used as a control. Mammary tissue biopsies were collected at times 0 and 3 h after infection. The total RNA was extracted and sequenced on an Illumina HiSeq 2000 (Illumina Inc., San Diego, CA). Transcripts were assembled from the reads aligned to the bovine UMD 3.1 reference genome, and the statistical analyses were performed using the previously mentioned tools (edgeR, baySeq, and Cuffdiff 2). Finally, the identified genes were submitted to pathway enrichment analysis. A total of 1,756, 1,161, and 3,389 genes with differential gene expression were identified when using edgeR, baySeq, and Cuffdiff 2, respectively. A total of 122 genes were identified by the overlapping of the 3 methods; however, only the platelet activation presented a significantly enriched pathway. From the results, we suggest the FCER1G, GNAI2, ORAI1, and VASP genes shared among the 3 methods in this pathway for posterior biological validation.

Composition of fatty acids, tocopherols, tocotrienols and ß-carotene content in oils of seeds of Brazilian Sapindaceae and Meliaceae species.

This study analyzes the lipid composition and the oxidative stability of oils of Sapindaceae and Meliaceae seeds. The oil content ranged from 14.7% (Guarea guidonia) to 30.1% (Allophylus puberulus and Paullinia elegans). Ten fatty acids were identified in seed oils. Guarea guidonia seeds accumulated mainly oleic (44.9%) and linoleic (24.1%) acid, whereas the unusual gondoic and paullinic acids were identified in A. puberulus (15.8%; 8.9%) and P. elegans (14.4%; 44.2%), respectively. The oil of P. elegans had the highest oxidative stability (16.2 h.). Tocopherol predominated in A. puberulus (10.4 mg/100 g) and tocotrienol predominated in G. guidonea and P. elegans (2.6 mg/100 g). The vitamers α-tocopherol, γ-tocopherol, and γ-tocotrienol were found in the studied oils. ß-carotene was predominantly detected in the oil of A. puberulus seeds (22.4 µg/g). Guarea guidonia seed oil has a high potential in food due to richness in essential fatty acids. In turn, A. puberulus and P. elegans oils could be suggested for other industrial purposes (e.g., biodiesel, varnishes, paints, soaps, or oleochemicals) due to their content of long-chain fatty acids.

Molecular identification and physiological characterization of Zymomonas mobilis strains from fuel-ethanol production plants in north-east Brazil.

Zymomonas mobilis has long attracted attention owing to its capacity to ferment hexose to ethanol. From a taxonomic viewpoint, Z. mobilis is a unique species of the genus Zymomonas, separated into three subspecies, Z. mobilis subsp. mobilis, Z. mobilis subsp. pomaceae and Z. mobilis subsp. francensis on the basis of physiological tests, which are often unreliable owing to the genetic proximity among these species. Currently, the use of molecular techniques is more appropriate for identification of these bacterial subspecies. In this study, the 32 strains of Z. mobilis present in the UFPEDA bacterial collection were characterized using molecular techniques, such as sequencing of the 16S rDNA gene and its theoretical restriction profile, classifying them as members of the subspecies, Z. mobilis subsp. mobilis. In addition, anaerobic cultivations were performed, which showed the biological diversity of the strains in terms of growth, sugar consumption and ethanol production. From these results, it was possible to identify the strain Z-2-80 as a promising bacterium for use in the fermentation process. SIGNIFICANCE AND IMPACT OF THE STUDY: Zymomonas mobilis is a bacterium of great relevance to biotechnology, owing to its capacity to ferment hexose to ethanol. On a molecular basis, 32 isolates were identified as Z. mobilis subsp. mobilis. However, intraspecific diversity was identified when these were grown under strictly anaerobic conditions. The results obtained from this study suggest a strain of Z. mobilis as an alternative for use in the fermentation process.

5-ALA-mediated photodynamic therapy reduces the parasite load in mice infected with Leishmania braziliensis.

Photodynamic therapy (PDT) has proven to be an effective alternative for the treatment of cutaneous leishmaniasis. Skin lesions consist of ulcers with well-defined raised edges, and granular floor. Th1 immune response is the protective profile in patients infected with Leishmania. In this study, the photodynamic therapy with 5-aminolevulinic acid, the parasitic load, and the modulation of the immune response was evaluated in mice infected with Leishmania braziliensis. Balb/c mice were infected with L. braziliensis and subsequently treated with three sections of PDT. The parasite load and mRNA expression of cytokines (IFN-γ, IL-4, IL-17, IL-22, IL-27, IL-10) and transcription factors (GATA-3, Foxp3 and T-bet) were analysed by quantitative PCR. The parasite load in the treated group was significantly lower than in the untreated group (P<.0001); in PDT treated animals, we observed an increase in IFN-γ and T-bet mRNA (P=.012 and P=.0071). There was a significant reduction in mRNA expression of IL-22 associated with an increased expression of IL-27 mRNA in the animals treated with light only (P=.0001). 5-ALA associated with photodynamic therapy promotes a reduction in parasite load and an increased expression of IFN-γ and T-bet mRNA.

Accuracy of genomic predictions in Gyr (Bos indicus) dairy cattle.

Genomic selection may accelerate genetic progress in breeding programs of indicine breeds when compared with traditional selection methods. We present results of genomic predictions in Gyr (Bos indicus) dairy cattle of Brazil for milk yield (MY), fat yield (FY), protein yield (PY), and age at first calving using information from bulls and cows. Four different single nucleotide polymorphism (SNP) chips were studied. Additionally, the effect of the use of imputed data on genomic prediction accuracy was studied. A total of 474 bulls and 1,688 cows were genotyped with the Illumina BovineHD (HD; San Diego, CA) and BovineSNP50 (50K) chip, respectively. Genotypes of cows were imputed to HD using FImpute v2.2. After quality check of data, 496,606 markers remained. The HD markers present on the GeneSeek SGGP-20Ki (15,727; Lincoln, NE), 50K (22,152), and GeneSeek GGP-75Ki (65,018) were subset and used to assess the effect of lower SNP density on accuracy of prediction. Deregressed breeding values were used as pseudophenotypes for model training. Data were split into reference and validation to mimic a forward prediction scheme. The reference population consisted of animals whose birth year was ≤2004 and consisted of either only bulls (TR1) or a combination of bulls and dams (TR2), whereas the validation set consisted of younger bulls (born after 2004). Genomic BLUP was used to estimate genomic breeding values (GEBV) and reliability of GEBV (R2PEV) was based on the prediction error variance approach. Reliability of GEBV ranged from ∼0.46 (FY and PY) to 0.56 (MY) with TR1 and from 0.51 (PY) to 0.65 (MY) with TR2. When averaged across all traits, R2PEV were substantially higher (R2PEV of TR1 = 0.50 and TR2 = 0.57) compared with reliabilities of parent averages (0.35) computed from pedigree data and based on diagonals of the coefficient matrix (prediction error variance approach). Reliability was similar for all the 4 marker panels using either TR1 or TR2, except that imputed HD cow data set led to an inflation of reliability. Reliability of GEBV could be increased by enlarging the limited bull reference population with cow information. A reduced panel of ∼15K markers resulted in reliabilities similar to using HD markers. Reliability of GEBV could be increased by enlarging the limited bull reference population with cow information.

Strategies for single nucleotide polymorphism (SNP) genotyping to enhance genotype imputation in Gyr (Bos indicus) dairy cattle: Comparison of commercially available SNP chips.

Genotype imputation is widely used as a cost-effective strategy in genomic evaluation of cattle. Key determinants of imputation accuracies, such as linkage disequilibrium patterns, marker densities, and ascertainment bias, differ between Bos indicus and Bos taurus breeds. Consequently, there is a need to investigate effectiveness of genotype imputation in indicine breeds. Thus, the objective of the study was to investigate strategies and factors affecting the accuracy of genotype imputation in Gyr (Bos indicus) dairy cattle. Four imputation scenarios were studied using 471 sires and 1,644 dams genotyped on Illumina BovineHD (HD-777K; San Diego, CA) and BovineSNP50 (50K) chips, respectively. Scenarios were based on which reference high-density single nucleotide polymorphism (SNP) panel (HDP) should be adopted [HD-777K, 50K, and GeneSeek GGP-75Ki (Lincoln, NE)]. Depending on the scenario, validation animals had their genotypes masked for one of the lower-density panels: Illumina (3K, 7K, and 50K) and GeneSeek (SGGP-20Ki and GGP-75Ki). We randomly selected 171 sires as reference and 300 as validation for all the scenarios. Additionally, all sires were used as reference and the 1,644 dams were imputed for validation. Genotypes of 98 individuals with 4 and more offspring were completely masked and imputed. Imputation algorithms FImpute and Beagle v3.3 and v4 were used. Imputation accuracies were measured using the correlation and allelic correct rate. FImpute resulted in highest accuracies, whereas Beagle 3.3 gave the least-accurate imputations. Accuracies evaluated as correlation (allelic correct rate) ranged from 0.910 (0.942) to 0.961 (0.974) using 50K as HDP and with 3K (7K) as low-density panels. With GGP-75Ki as HDP, accuracies were moderate for 3K, 7K, and 50K, but high for SGGP-20Ki. The use of HD-777K as HDP resulted in accuracies of 0.888 (3K), 0.941 (7K), 0.980 (SGGP-20Ki), 0.982 (50K), and 0.993 (GGP-75Ki). Ungenotyped individuals were imputed with an average accuracy of 0.970. The average top 5 kinship coefficients between reference and imputed individuals was a strong predictor of imputation accuracy. FImpute was faster and used less memory than Beagle v4. Beagle v4 outperformed Beagle v3.3 in accuracy and speed of computation. A genotyping strategy that uses the HD-777K SNP chip as a reference panel and SGGP-20Ki as the lower-density SNP panel should be adopted as accuracy was high and similar to that of the 50K. However, the effect of using imputed HD-777K genotypes from the SGGP-20Ki on genomic evaluation is yet to be studied.

Chicken embryo: a useful animal model for drug testing?

The use of chicken embryos (CEs) as an in vivo experimental model for different pharmaceutical purposes is not a novelty. However, in recent years, the number of reports employing CE to evaluate several parameters, such as the toxicity and efficacy of drugs and/or nanosystems, has increased. Therefore, this review discusses the relevance of CE for drug testing, emphasizing the inoculation routes and the embryonic stages. The challenges to be overcome, as well as some practical recommendations to allow CE to be more explored as a promising in vivo model in drug analyses, are also highlighted.

Quantitative trait loci affecting milk production traits on bovine chromosome 6 in zebuine Gyr breed.

Fourteen Brazilian dairy Gyr sire families with 657 daughters were analyzed for quantitative trait loci (QTL) on chromosome 6 by using a daughter design for 5 economic traits: milk, fat, and protein production, fat and protein percentage. The cows and sires were genotyped for 27 microsatellites with average spacing between markers of 4.9 cM. In the analyses across 14 families, for the largest significant families, and within family, a QTL was located for milk yield and fat yield close to marker BMS2508 at the 5% chromosome-wide significance level across families and 1% chromosome-wide within families. For fat percentage, a QTL near DIK4482 was identified at the 5% chromosome-wide significance level when all families were analyzed together and at the 1% chromosome-wide significance level within the largest significant families. The different analyses yielded results that were generally consistent for milk yield, fat yield, and fat percentage. The order of the markers in the derived map was consistent with that in the consensus map. Some QTL and candidate genes in dairy cattle for milk production traits are probably preserved in Bos taurus and Bos indicus.

Anthelmintic activity of Azadirachta indica A. Juss against sheep gastrointestinal nematodes.

Gastrointestinal nematode control has been performed through use of anthelmintics. However, the development of resistant populations has required research into new alternatives. There are popular reports about anti-parasitic activity of Azadirachta indica in animals and plants. The aim of this study was to evaluate the anthelmintic activity of A. indica after feeding sheep with the dried leaves. In this experiment, 40 sheep were allotted into four treatment groups. Group I received a treatment of A. indica dry leaves mixed in a concentrate at a rate of 0.1 g/kg dose for 3 months. Group II was treated with double the dose of Group I. Group III was treated with closantel (Diantel) at the manufacturer-recommended dose once at the beginning of the study and Group IV was not treated. To compare treatment effects, the following parameters were evaluated: egg count per gram of feces (EPG), worm burden, weight gain and haematocrit. EPG and worm burden results were statistically evaluated using the Kruskal-Wallis test. Haematocrit and live weight gain were submitted to analysis of variance (ANOVA) and the means evaluated by Tukey's test with 95% probability. None of the evaluated parameters of the treatment groups were statistically different when compared to the control group, demonstrating that, with the protocol used, A. indica has no anthelmintic effect.

Immunoblot with cerebrospinal fluid from patients with neurocysticercosis using antigen from cysticerci of Taenia solium and Taenia crassiceps.

A comparative study was conducted on membrane (M) and vesicular fluid (VF) from cysticerci of Taenia solium (Tso) obtained from naturally infected swine and the Taenia crassiceps ORF strain (Tc) maintained by experimental infection of female BALB/c mice. The study was carried out using immunoblotting to detect antibodies in cerebrospinal fluid (CSF) from patients with neurocysticercosis. No reactivity was observed in the 32 samples from a control group. Of the 23 CSF fluid samples from patients with neurocysticercosis, 22 (95.6%) were reactive in the M-Tso blot and 21 (91.3%) were reactive in the other three blots (VF-Tso, M-Tc, and VF-Tc). Immunodominant peptides in each antigen were 98-92 kD, 56-52 kD, and 72-68 kD in M-Tso; 72-68 kD, 120 kD, 155 kD, 98-94 kD, 76 kD, and 115-108 kD in VF-Tso: 72 kD, 62 kD, and 42 kD in M-Tc; and 72-68 kD and 95-92 kD in VF-Tc. The cross-reactivity observed in the immunoblots performed on CSF samples from patients with neurocysticercosis indicates that the parasites share important epitopes present at sufficient concentrations for use in immunologic tests.

Immunodiagnosis of human leptospirosis by dot-ELISA for the detection of IgM, IgG, and IgA antibodies.

A dot-ELISA was evaluated using antigen obtained from Leptospira interrogans cultures of the serovars brasiliensis, canicola, cynopteri, hebdomadis, and icterohaemorrhagiae for the detection of human IgM, IgG, and IgA. Single serum samples from 63 patients with the icterohemorrhagic form of leptospirosis in the acute phase, collected 3-14 days (mean = 7 days) after the onset of symptoms were tested. Ten patients were examined during convalescence and followed up for a period of 4-12 months. For a control group, serum samples from 10 apparently healthy individuals with no clinical or epidemiologic history of leptospirosis, and from 38 patients with nonleptospiral illnesses were used. In the acute phase, IgM antibodies were detected in 62 (98%) of 63 patients and IgG and IgA were observed in 70% and 76% of them, respectively. For the admission serum samples, the predictive value negative of the dot-ELISA was 98% for IgM, 72% for IgG, and 76% for IgA detection. All 10 patients followed-up during convalescence showed IgM antibodies up to the sixth month, decreasing to 57% by the 10th month, and persisting in only one of six patients during the 11th and 12th months of follow-up. Immunoglobulin G was detected in six patients up to the fourth month and in two of six individuals up to the end of follow-up. Immunoglobulin A was observed in all patients up to the end of the first month, decreasing progressively up to the sixth month, and was no longer detected in any patients from seventh to the 12th months of follow-up. The dot-ELISA can be used as an important laboratory screening test, especially when detecting IgM antibodies. It proved to be effective in the diagnosis of human leptospirosis, and appears to have advantages in terms of yield, time, and case of execution and low cost.

Immunodiagnosis of human leptospirosis using saliva.

When the enzyme-linked immunosorbent assay for the detection of specific immunoglobulin M class antibodies was applied to paired saliva and serum samples from 40 patients with leptospirosis, positivity was 87.5% and 100%, respectively. No positive result was obtained with any saliva or serum sample from 60 individuals used as controls. These results suggest the alternative use of saliva for diagnosis and for epidemiological studies of human leptospirosis.

It is valid to dilate native aortic coarctation with a balloon catheter.

We report our experience in 37 patients with aortic coarctation who underwent balloon aortoplasty. Of the 37, the lesion was native in 34 cases. Clinical re-evaluation was possible in 22 patients, with a mean follow-up period of 13 months and, of these, 21 patients were asymptomatic. Haemodynamic and angiographic studies were performed in 13 patients, with a mean period of follow-up of 12 months, showing excellent results in 11 patients. Aneurysmal formation had occurred in one and recoarctation in the other patient. Our experience has proved that balloon aortoplasty is an effective method for treating patients with aortic coarctation. Adequate selection in terms of the morphology of the coarctation and the size of the balloon catheter are crucial factors in the success of the procedure.

Ocular manifestations in the acute phase of leptospirosis.

OBJECTIVES: To study the ocular manifestations during the acute phase of leptospirosis and their incidence in hospitalized patients due to systemic complications, and to verify the importance of routine ophthalmologic care in these cases. METHODS: Twenty-one patients, 20 males and 1 female, with clinical and laboratory (ELISA IgM) diagnoses of leptospirosis were subjected to ophthalmologic examination. RESULTS: We observed conjunctival hyperemia in 18 patients (85.7%), increased retinal venous caliber in 12 patients (57.1%), optic disc redness in 12 patients (57.1%), subconjunctival hemorrhage in four patients (19.0%), optic disc edema in one patient (4.8%), retinal vasculitis in one patient (4.8%), retinal hemorrhage in one patient (4.8%), hard exudates in one patient (4.8%), and papillitis in one patient (4.8%). No anterior chamber reaction was found. CONCLUSIONS: We observed a high incidence of several ocular manifestations in the acute phase of leptospirosis. Despite the systemic severity and high incidence of ocular disorders in the acute phase of leptospirosis, the short-term visual outcome of these patients was good.

[An immunoenzyme test (ELISA) for the detection of circulating class-IgA antibodies in human leptospirosis]. / Teste imunoenzimático (ELISA) para detecção de anticorpos circulantes da classe IgA na leptospirose humana.

An enzyme-linked immunosorbent assay ELISA was evaluated for the detection of IgA antibodies in the human leptospirosis. The assay proved to be sensitive and specific when compared with the ELISA-IgM, in the examined serum samples. The results found suggest that IgA antibodies became positive later in leptospirosis, and will can be an evolutive indicator in the development of the disease.