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Background: The growing contact between men and wild animals, caused by the increase in the population in urban centers and the destruction of the habitat of these animals, has been leading to a greater circulation of pathogens between humans and wildlife. Chelonoidis carbonaria, a tortoise found throughout South America, is one of the animals most rescued from animal trafficking and illegal breeding. Considering this situation, this study aimed to verify the occurrence of hemoparasites in C. carbonaria. Materials and Methods: Blood samples from 73 C. carbonaria were collected from animals located in (1) a rural commercial breeding unit, (2) an urban zoo, and (3) a center of rescued animal screening. Genomic DNA was extracted from these animals and used in PCRs to detect specific genomic fragments of haemogregarines (i.e., Hepatozoon and Hemolivia), and members of the Anaplasmataceae Family (i.e., Ehrlichia sp. and Anaplasma sp.). Blood samples were screened for hemopathogens by direct microscopy and were used for hematological assays, and serum samples were analyzed to determine the concentration of serum components. Results: It was found that 34.2% of the tortoises presented Sauroplasma sp. in their blood samples; these animals showed clinical biochemistry changes that indicate altered liver function. Two zoo animals were positive for Ehrlichia sp. in PCR, and also presented clinical biochemistry and hematological changes. Conclusion: The present project is pioneer in the detection of Ehrlichia sp. in C. carbonaria, and was able to identify changes in clinical biochemistry that can be a result of the infection by hemopathogens in this species.

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Dental cements are widely used in the clinical routine, specifically for root canal sealing. Within this context, it is expected that these materials present antimicrobial activity, since it would help in the prevention of apical and periapical infections. The present study aimed to comparatively verify the antimicrobial activity of four dental cements against microorganisms that are routinely isolated from endodontic infections. Reference strains of Enterococcus faecalis, Candida albicans and Escherichia coli were submitted to the agar diffusion test and to modified direct contact test using four different sealers: an eugenol zinc oxide compound, an epoxy resin associated to calcium hydroxide and bismuth, a mineral trioxide aggregate (MTA) and a bioceramics. Different E. coli, C. albicans and E. faecalis growth inhibition profiles were observed in the agar diffusion assay. In the direct contact test, the bioceramics presented a higher microbicide activity on all microorganisms tested herein. Dental cements have different antimicrobial activities, being that the bioceramics present the most consistent antimicrobial activity, and that the direct contact test presented more uniform results than the agar diffusion test. This study reveals the antimicrobial activities of different cements and allow dentists to decide which material to employ in their daily practice.

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This prospective study aimed to evaluate the effect of metronomic cyclophosphamide on carboplatin's tolerability, efficacy, and pharmacokinetics in dogs with mammary carcinoma. Sixteen female dogs with mammary carcinoma were divided into groups: 300 mg/m2 intravenous (i.v.) carboplatin therapy (G1 = 8) or 300 mg/m2 i.v. carboplatin which was associated with 12.5 mg/m2 oral cyclophosphamide in a metronomic regimen (G2 = 8). The investigated animals underwent a clinical evaluation, a mastectomy, a carboplatin chemotherapy, and serial blood sampling for the pharmacokinetic analysis. The adverse events and survival rates were monitored. A non-compartmental analysis was applied to calculate the pharmacokinetic parameters of carboplatin in the 2nd and 4th chemotherapy cycles. Carboplatin PK showed high interindividual variability with a 10-fold variation in the area under the plasma concentration−time curve (AUC) in G1. The systemic plasma exposure to carboplatin was equivalent in both of the treatments considering the AUC and maximum plasma concentration (Cmax) values. Although the red blood cells (p < 0.0001), platelets (p = 0.0005), total leukocytes (p = 0.0002), and segmented neutrophils (p = 0.0007) were reduced in G2, the survival rate increased (p = 0.0044) when it was compared to G1. In conclusion, adding low daily doses of cyclophosphamide to a carboplatin therapy showed promising outcomes in female dogs with mammary tumors.

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Biochemical, serological, and molecular methods have been developed for the laboratory diagnosis of diseases caused by C. pseudotuberculosis (CP), but the identification of the pathogen and biovars differentiation may be time-consuming, expensive, and confusing compared with other bacteria. This study aimed to evaluate MALDI Biotyper and Overall Genome Relatedness Index (OGRI) analysis to optimize the identification and differentiation of biovars of C. pseudotuberculosis. Out of 230 strains isolated from several hosts and countries, 202 (87.8%) were precisely classified using MALDI Biotyper and the BioNumerics platform. The classification accuracies for the Ovis and Equi biovars were 80 (88.75%) and 82 (92.68%), respectively. When analyzing a sampling of these strains by Average Nucleotide Identity based on BLAST and TETRA analyses using genomic sequence data, it was possible to differentiate 100% of the strains in Equi and Ovis. Our data show that MALDI Biotyper and OGRI analysis help identify C. pseudotuberculosis at the species and biovar levels.

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Toxocariasis is an infection caused by the round worms Toxocara canis and Toxocara cati. It occurs worldwide though it is more prevalent in developing countries. For the diagnosis of toxocariasis, the most used method is the indirect enzyme-linked immunosorbent assay (indirect ELISA), based on the detection of specific antibodies using the excreted/secreted products from T. canis larvae (TES) as antigens, but it cross-reacts with several helminth infections. For this reason, there is a need to investigate species-specific immunoreactive proteins, which can be used for the development of a more sensitive and specific diagnosis. This study aims to investigate immunoreactive protein candidates to be used for the development of a more sensitive and specific diagnosis of Toxocara spp. infection in humans. We have used immunoblotting and mass spectrometry to select four Toxocara canis immunoreactive proteins that were recombinantly expressed in bacteria and evaluated as potential new diagnostic antigens (rMUC3, rTES 26, rTES32 and rCTL4). The recognition of these recombinant proteins by total serum IgG and IgG4 was assayed using the purified proteins in an isolated manner or in combination. The IgG ELISAs performed with individual recombinant antigens reached values of sensitivity and specificity that ranged from 91.7% to 97.3% and 94.0% to 97.9%, respectively. Among the analyses, the IgG4 immunoassay was proven to be more effective, revealing a sensitivity that ranged from 88.8% to 98.3% and a specificity of 97.8%-97.9%. The IgG4 ELISA was shown to be more effective and presented no cross-reactivity when using combinations of the rTES 26 and rCTL4 recombinant proteins. The combination of these two molecules achieved 100% sensitivity and specificity. The use of only two recombinant proteins can contribute to improve the current panorama of toxocariasis immunodiagnosis for, with a better optimization and reduced cost.

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Candida spp. resistant to commercially available antifungals are often isolated from patients with oral candidiasis, a situation that points to the need for the development of new therapies. Thus, we evaluated the activity of Fusarium oxysporum-based silver nanoparticles (AgNPs) on Candida spp. isolated from denture stomatitis lesions. Candida isolates were molecularly identified and submitted to susceptibility assays using AgNPs and commercial fungicides. The interference on biofilm formation and the mechanisms of action of AgNPs on Candida spp. were also investigated. Scanning electron microscopy was used to evaluate the morphology of AgNP-treated Candida. Candida albicans was the most frequent species isolated from denture stomatitis cases. All Candida spp. were susceptible to AgNPs at low concentrations, except Candida parapsilosis. AgNPs caused surface damage, cell disruption, and biofilm formation inhibition. The ergosterol supplementation protected C. albicans against the AgNP action. AgNPs are effective against Candida spp. and can be faced as a promising new therapeutic agent against oral candidiasis.

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Caseous Lymphadenitis (CLA) is an infectious disease that affects small ruminants, and the bacterium Corynebacterium pseudotuberculosis is its etiologic agent. This disease presents a high morbidity and a great economic impact on goat farming, leading to reduced milk and meat production and reproductive losses. The available data about the influence of C. pseudotuberculosis on the reproductive system were obtained after experimental inoculations. In this way, this study aimed to evaluate the influence of the natural infection by C. pseudotuberculosis on the luteal function and serum progesterone (P4) levels in goats. Sixteen female goats were diagnosed for the presence of C. pseudotuberculosis specific antibodies and divided into 2 groups: CLA positive and negative animals. They were submitted to estrous synchronization, followed by controlled mating. Corpus luteum and serum levels of P4 were evaluated on the 7th and 20th days after mating (D7 and D20). The ultrasonographic results were not statistically different between the 2 groups, as well as serum P4 levels on D7. However, a significant increase in serum P4 levels on the CLA positive group was identified on D20, along with a significant correlation between C. pseudotuberculosis specific antibody production, and P4 serum levels. The result of this study indicates that the infection by C. pseudotuberculosis may influence the reproductive status of female goats through an enhanced production of progesterone.

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The aims of the present study were (i) to genotype Corynebacterium pseudotuberculosis, C. silvaticum, and C. auriscanis strains using enterobacterial repetitive intergenic consensus (ERIC-PCR), and (ii) to analyze the epidemiological relationships among isolates according to biovar (Equi and Ovis), species, host, and geographical origin of the C. pseudotuberculosis strains. Sixty-eight C. pseudotuberculosis, nine C. silvaticum, and one C. auriscanis, C. pseudotuberculosis ATCC® 19410™ strain and the attenuated C. pseudotuberculosis 1002 vaccinal strain were fingerprinted by ERIC 1+2-PCR. Field strains were isolated from various hosts (cattle, buffaloes, sheep, goats, horses, dogs, and pigs) in six countries (Mexico, Portugal, Brazil, Equatorial Guinea, Egypt, and Israel). High genetic diversity was found among the studied Corynebacterium spp. isolates, clustering in 24 genotypes with a Hunter & Gaston diversity index (HGDI) of 0.937. The minimal spanning tree of Corynebacterium spp. revealed three clonal complexes, each associated with one bacterial species. Twenty-two genotypes were observed among C. pseudotuberculosis isolates, with an HGDI of 0.934. Three major clonal complexes were formed at the minimal spanning tree, grouped around the geographic origin of C. pseudotuberculosis isolates. These results reinforce the high typeability, epidemiological concordance, and discriminatory power of ERIC-PCR as a consistent genotyping method for C. pseudotuberculosis, which could be useful as an epidemiological tool to control caseous lymphadenitis. Moreover, our results also indicate the potential of ERIC 1+2-PCR for the genotyping of other species of Corynebacterium other than C. pseudotuberculosis.

Os objetivos do presente estudo foram (i) genotipar amostras de Corynebacterium pseudotuberculosis, C. silvaticum e C. auriscanis usando Enterobacterial Repetitive Intergenic Consensus (ERIC-PCR), bem como (ii) analisar as relações epidemiológicas entre os isolados de acordo com biovar (Equi e Ovis), espécie, hospedeiro e origem geográfica das amostras de C. pseudotuberculosis. Sessenta e oito isolados de C. pseudotuberculosis, nove C. silvaticum, um C. auriscanis, C. pseudotuberculosis ATCC® 19410 ™ e a amostra vacinal atenuada C. pseudotuberculosis 1002 foram tipificadas por ERIC 1 + 2-PCR. As amostras de campo foram isoladas de diferentes hospedeiros (bovinos, búfalos, ovinos, caprinos, equinos, cães e suínos) em seis países (México, Portugal, Brasil, Guiné Equatorial, Egito e Israel). Uma alta diversidade genética foi observada entre os isolados de Corynebacterium spp., agrupados em vinte e quatro genótipos com um índice de diversidade Hunter & Gaston (HGDI) de 0,937. A análise da minimal spanning tree (MST) de Corynebacterium spp. revelou três complexos clonais, cada um associado a uma espécie bacteriana. Vinte e dois genótipos foram observados entre isolados de C. pseudotuberculosis, com um HGDI de 0,934. Na análise da MST, três grandes complexos clonais foram formados, agrupando-se em torno da origem geográfica dos isolados de C. pseudotuberculosis. Esses resultados reforçam a alta tipabilidade, concordância epidemiológica e poder discriminatório do ERIC-PCR como método consistente de genotipagem para C. pseudotuberculosis, podendo ser útil como ferramenta epidemiológica no controle da linfadenite caseosa. Além disso, os resultados também indicam o grande potencial de ERIC 1 + 2-PCR para genotipagem de espécies do gênero Corynebacterium além de C. pseudotuberculosis.

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Silver nanoparticles (AgNPs) have been successfully applied in several areas due to their significant antimicrobial activity against several microorganisms. In dentistry, AgNP can be applied in disinfection, prophylaxis, and prevention of infections in the oral cavity. In this work, the use of silver nanoparticles in dentistry and associated technological innovations was analyzed. The scientific literature was searched using PubMed and Scopus databases with descriptors related to the use of silver nanoparticles in dentistry, resulting in 90 open-access articles. The search for patents was restricted to the A61K code (International Patent Classification), using the same descriptors, resulting in 206 patents. The results found were ordered by dental specialties and demonstrated the incorporation of AgNPs in different areas of dentistry. In this context, the search for patents reaffirmed the growth of this technology and the dominance of the USA pharmaceutical industry over AgNPs product development. It could be concluded that nanotechnology is a promising area in dentistry with several applications.

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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.