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Staphylococcus pseudintermedius is frequently associated with several bacterial infections in dogs, highlighting a One Health concern due to the zoonotic potential. Given the clinical significance of this pathogen, we performed comprehensive genomic analyses of 28 S. pseudintermedius strains isolated from canine infections throughout whole-genome sequencing using Illumina HiSeq, and compared the genetic features between S. pseudintermedius methicillin-resistant (MRSP) and methicillin-susceptible (MSSP) strains. Our analyses determined that MRSP genomes are larger than MSSP strains, with significant changes in antimicrobial resistance genes and virulent markers, suggesting differences in the pathogenicity of MRSP and MSSP strains. In addition, the pangenome analysis of S. pseudintermedius from canine and human origins identified core and accessory genomes with 1847 and 3037 genes, respectively, which indicates that most of the S. pseudintermedius genome is highly variable. Furthermore, phylogenomic analysis clearly separated MRSP from MSSP strains, despite their infection sites, showing phylogenetic differences according to methicillin susceptibility. Altogether our findings underscore the importance of studying the evolutionary dynamics of S. pseudintermedius, which is crucial for the development of effective prevention and control strategies of resistant S. pseudintermedius infections.

ABSTRACT: Surface contamination by pathogenic and deteriorating microorganisms is a constant concern in the food industry. Therefore, in this study we evaluated the antimicrobial activity of silver nanoparticles (AgNPs), produced using the essential oil of Lippia origanoides Kunth and ethanolic extract of Hymenaea martiana Hayne, against bacteria used in milk processing. For this, AgNPs were synthesized and their antimicrobial activity was evaluated for minimum inhibitory and bactericidal concentrations, time-kill, interference on the biofilm, and application on the surfaces of different materials. AgNPs showed bacteriostatic and bactericidal activity against Staphylococcus aureus (ATCC 25923), Staphylococcus aureus (ATCC 33591), Escherichia coli (ATCC 25922), Salmonella enterica subsp. enterica serovar Choleraesuis (ATCC 10708), Listeria monocytogenes, Escherichia coli, Salmonella spp.,andPseudomonas aeruginosa,and interfered with biofilm formation more than with consolidated biofilms. A 1hour period was sufficient to reduce the bacterial cells, whereas a Minimum Duration for Killing (MDK) of 99% was reached after approximately 30 min. AgNPs were effective against bacteria attached to stainless steel and polyethylene, but ineffective on tile surfaces. Thus, owing to the growing microbial resistance and the need to develop new products based on the concepts of green chemistry, these AgNPs are presented as a new possibility for cleaning processes in the food industry.

The development of alternative (and free-of-antibiotics) antibacterial and antibiofilm agents is an important strategy to circumvent the resistance of bacteria to antibiotics. Herein, we explore the production of mixed oxides by incorporating silver nanoparticles in titanium dioxide as a silver concentration-dependent antibacterial agent that is further incorporated in Tilapia fish skin (a promising prototype of xenograft), integrating the antibacterial activity of mixed oxide into the intrinsic properties of Tilapia skin. The antibiofilm activity of samples prepared with high concentrations of silver (10 wt% of precursor AgNO3) has been considered a good antibiofilm response. The influence of silver content is also observed with respect to the minimum bactericidal concentration, which is reduced to 3.13 mg/mL with a characteristic kill time in the order of 30 min that is associated with antibiofilm activity in biofilm-forming strains of Staphylococcus aureus. These results indicate that modified Tilapia fish skin acquires antibacterial behavior and can be explored for xenografts with prospective applications in the light-dependent actuation of TiO2-based compounds.

The development of composites with antibacterial activity represents an important strategy to avoid side effects such as increasing bacterial resistance to antibiotics. In particular, the green synthesis of metal nanoparticles avoids the use of hazardous chemical compounds and introduces the intrinsic beneficial properties of plant-derived compounds. Herein, the reduction of gold salt into metal nanoparticles was provided by the action of a cationic polymer derived from tannin (Tanfloc®). Comparative activity of antibacterial agents (pure Tanfloc and Au NPs—Tanfloc) at different concentrations were evaluated in terms of the antibiofilm activity, kill-time assays and inhibition haloes confirming the antibacterial activity of the Tanfloc that is reinforced by the incorporation of reduced gold nanoparticles, resulting in the complete elimination of S. aureus from an initial concentration of 108 CFU/mL after 120 min of reaction of Au NPs + Tanfloc solution in association with strong inhibition of the biofilm formation attributed to the Tanfloc.

Objetivo: analisar o potencial de formigas como veículos mecânicos de bactérias do gênero Staphylococcus aureus no centro cirúrgico e unidade de terapia intensiva de um hospital público. Metodologia: trata-se de um estudo descritivo. As formigas foram coletadas utilizando-se iscas atrativas que ficavam expostas por três horas sendo, após a exposição, incubadas por 24h a 35ºC. A susceptibilidade aos antimicrobianos foi analisada através da técnica de disco-difusão. Os isolados de Cocos gram-positivos foram submetidos a Reação em Cadeia da Polimerase e observado os genes nuc, mecA blaZ e icaAD. Resultados: foram coletadas 592 formigas pertencentes à espécie Tapinoma melanocephalum e Pheidole sp. Todos os isolados apresentaram o gene mecA, 93,7% apresentaram resultado positivo para o gene blaZ e 12,5% para o gene icaA e icaD. Sete isolados apresentaram multirresistência. Conclusão: os resultados sugerem que as formigas podem agir como veiculadoras de bactérias multirresistentes no ambiente hospitalar. Abstract Objective: to analyze the potential of ants as mechanical vehicles of bacteria of the genus Staphylococcus Aureus in the surgical center and intensive care unit of a public hospital. Methodology: this was a descriptive study. The ants were collected using attractive baits that were exposed for three hours and, after exposure, incubated for 24 hours at 35ºC. The susceptibility to antimicrobials was analyzed using the disk-diffusion technique. Gram-positive Cocos isolates were subjected to Polymerase Chain Reaction and the nuc, mecA blaZ and icaAD genes were observed. Results: 592 ants belonging to the species Tapinoma melanocephalum and Pheidole sp. All isolates showed the mecA gene, 93.7% were positive for the blaZ gene and 12.5% for the icaA and icaD gene. Seven isolates showed multidrug resistance. Conclusion: the results suggest that ants can act as carriers of multidrug-resistant bacteria in the hospital environment.

Over the last few decades, diseases have emerged as a major bottleneck in fish farming, especially with the emergence of several fish pathogens and increasing resistance to treatments [...]

RESUMO: Considerando a necessidade de terapias alternativas para a mastite bovina, uma inflamação, normalmente de causa infecciosa, de alta prevalência e de alto impacto econômico nas fazendas leiteiras, avaliou-se a eficácia antimicrobiana, in vivo, de uma formulação intramamária à base de polipirrol (PPy) solúvel, um polímero condutor promissor para aplicações biomédicas, especialmente como potente antibacteriano. Neste ensaio, quartos mamários de vacas holandesas sadias (n = 8) foram inoculados com Staphylococcus aureus e tratados, por via intramamária, com formulação experimental à base de PPy solúvel (5%) e com formulação comercial à base de sulfato de gentamicina. O efeito desses tratamentos foi avaliado com a realização de lactoculturas, da Contagem Bacteriana Total (TBC), da Contagem de Células Somáticas (SCC) e da análise da composição do leite das amostras obtidas de quartos mamários, em sete momentos experimentais. Avaliação hematológica dos animais também foi realizada. A aplicação intramamária de três doses da formulação experimental à base de PPy solúvel resultou em maiores log/mL da TBC e da SCC quando comparadas ao grupo controle positivo e ao grupo que recebeu sulfato de gentamicina. A administração da formulação experimental não induziu alterações na composição do leite e nos parâmetros hematológicos. Alguns fatores farmacocinéticos e farmacodinâmicos do PPy solúvel podem ser atribuídos a ineficácia antimicrobiana da pomada experimental. Outras pesquisas devem ser realizados em prol do desenvolvimento de formulações que permitam a atuação antibacteriana do PPy solúvel no ambiente intramamário de vacas leiteiras.

Antimicrobial resistance is increasing despite new treatments being employed, so novel strategies are required to ensure that bacterial infections remain treatable. Bacteriophages (phages; bacteria viruses) have the potential to be used as natural antimicrobial methods to control bacterial pathogens such as Salmonella spp. A Salmonella phage, Wara, was isolated from environmental water samples at the Subaé River Basin, Salvador de Bahia, Brazil. The basin has environmental impacts in its main watercourses arising from the dumping of domestic and industrial effluents and agricultural and anthropological activities. The phage genome sequence was determined by Oxford Nanopore Technologies (ONT) MinION and Illumina HiSeq sequencing, and assembly was carried out by Racon (MinION) and Unicycler (Illumina, Illumina + MinION). The genome was annotated and compared to other Salmonella phages using various bioinformatics approaches. MinION DNA sequencing combined with Racon assembly gave the best complete genome sequence. Phylogenetic analysis revealed that Wara is a member of the Tequintavirus genus. A lack of lysogeny genes, antimicrobial resistance, and virulence genes indicated that Wara has therapeutic and biocontrol potential against Salmonella species in healthcare and agriculture.

Aeromonas veronii is a Gram-negative bacterial species that causes disease in fish and is nowadays increasingly recurrent in enteric infections of humans. This study was performed to characterize newly sequenced isolates by comparing them with complete genomes deposited at the NCBI (National Center for Biotechnology Information). Nine isolates from fish, environments, and humans from the São Francisco Valley (Petrolina, Pernambuco, Brazil) were sequenced and compared with complete genomes available in public databases to gain insight into taxonomic assignment and to better understand virulence and resistance profiles of this species within the One Health context. One local genome and four NCBI genomes were misidentified as A. veronii. A total of 239 virulence genes were identified in the local genomes, with most encoding adhesion, motility, and secretion systems. In total, 60 genes involved with resistance to 22 classes of antibiotics were identified in the genomes, including mcr-7 and cphA. The results suggest that the use of methods such as ANI is essential to avoid misclassification of the genomes. The virulence content of A. veronii from local isolates is similar to those complete genomes deposited at the NCBI. Genes encoding colistin resistance are widespread in the species, requiring greater attention for surveillance systems.

Mastitis, mainly caused by bacterial intramammary infections, is the main problem in the breeding of dairy animals. The inflammations of the mammary gland is separated by types of mastitis, being subclinical, clinical, and the most severe, gangrenous mastitis. Here, we used 16S rRNA amplicon sequencing to characterize the bacterial microbiota of goat milk in the different types of goat mastitis caused by bacteria. We used 72 goat milk samples from a region of the state of Minas Gerais in Brazil, of which 12 were from clinically healthy animals, 42 from animals diagnosed with subclinical mastitis, 16 from animals with clinical mastitis, and 2 from animals with gangrenous mastitis. The group related to gangrenous mastitis was the most divergent in terms of alpha and beta diversity. The most abundant genus among samples of the groups was Staphylococcus spp., and we found a high abundance of Mycoplasma sp. in the milk of animals diagnosed with clinical mastitis. The most statistically relevant microorganisms among the groups were Prevotella sp., Ruminococcaceae, Prevotella ruminicola sp., and Providencia sp. We highlight a new association of bacterial agents in gangrenous mastitis among Escherichia sp./Shigella sp. and Enterococcus sp. and provide the second report of the genus Alkalibacterium sp., in milk samples. Only the taxa Staphylococcus sp., Bacteroides sp., Enterococcus, and Brevidabacterium sp., were present in all groups. The superpathway of L-tryptophan biosynthesis metabolites and the sucrose degradation III (sucrose invertase) pathway were the most prominent ones among the groups. In this study, we demonstrate how a rich microbiota of goat milk from healthy animals can be altered during the aggravation of different types of mastitis, in addition to demonstrating new bacterial genera in milk not previously detected in other studies as well as new associations between agents.

The present study aimed to assess the prevalence and risk factors associated with bovine brucellosis in slaughterhouses in the state of Ceará using spatial distribution and flow network analysis. Four slaughterhouses were sampled in Ceará: two under municipal inspection and two under state inspection. Blood samples were randomly collected from bovine animals, resulting in a total of 964 samples. The collected sera were subjected to the Acidified Buffered Antigen (AAT) test, and the complement fixation test (FC) was performed for positive cases. An epidemiological questionnaire was applied to 38 producers who slaughter animals at the sampled facilities to assess the risk factors for brucellosis. An apparent prevalence of 1.55% (15) was found in the AAT test and 0.2% (n=2) in the FC test. A higher percentage of reactive animals was observed (66.6%) in properties where cattle farming is not the main activity, with OR = 4.75. The absence of contact with neighboring animals is a factor associated with protection, with a lower prevalence of seroreactive animals (23.5%) when animals were raised without contact with others (OR = 0.30). Therefore, bovine brucellosis in herds and animals can be considered low in the studied region and under all production systems. Nevertheless, despite the importance of this disease to the economic and public health aspects and the advances of the PNCEBT Program, brucellosis is still circulating in Ceará.

Abstract Background Staphylococcus aureus is one of the main causative agents of mastitis in small ruminants. Antimicrobial use is the major treatment, but there are many flaws linked to resistance, tolerance or persistence. This study aimed to verify changes in resistance, virulence and clonal profiles of S. aureus isolated from persistent mastitis goat milk before and after enrofloxacin treatment. Results MIC increased to at least one antimicrobial in S. aureus isolates after enrofloxacin treatment compared to before. The most detected resistance genes before and after treatment were tetK, tetM, and blaZ, with more resistance genes detected after enrofloxacin treatment (p

ABSTRACT: The goat milk industry has unquestionably grown in recent years due to the high demand for dairy products, which are considered nutritious and hypoallergenic. As a result, security measures are required in the production chain to provide consumers with safe products, although the concept of biosecurity is still incipient in Brazilian goat farming. Therefore, this study aimed to review the main biosecurity measures applied to dairy goat farms and suggest a program that contemplates these measures to promote animal health and welfare, given that biosecurity bolsters environmental sustainability and improves this agricultural sector. Biosecurity programs (BP) are composed of a set of measures and procedures aimed at herd health and applied in all stages of animal husbandry, interacting with different sectors that make up the production system and providing a set of policies and operational norms to protect herds against potentially pathogenic agents. Thus, BP require planning, execution, monitoring, audits, updates, understanding of the procedures, and awareness of those involved in the production chain. In addition, it includes continuing education programs and the development of contingency plans for specific emergencies. The information included in this study provides input to dairy goat farms to implement practices that improve the productivity of this agricultural sector.

ABSTRACT: The purpose of this review was to address the applicability of nanotechnology in veterinary medicine, with an emphasis on research in Brazil from 2013 to 2020. Firstly, we introduced to the general aspects of applicability of nanotechnology in veterinary medicine, and lately we pointed the research involving nanoscience performed in Brazil, in the studied period. Nanotechnology is the field of science that has the capacity to organize matter in nanoscale structures (1 to 100 nm), enabling innovations in different areas including biotechnology, agriculture, disease diagnosis, food and clothing industry, electronics, and pharmacological therapies. In veterinary medicine, several studies are being carried out in the world, mainly in the areas that involve search of new treatment options and the development of immunotherapy, as well as in the diagnosis of diseases. In Brazil, it is clear that the use of nanotechnology in veterinary medicine is still incipient, but it can be considered a growing area. In addition, several points have to be reflected and researched, including some adverse effects and implications to validate the safe use of nanotechnology in veterinary medicine. Therefore, this review highlighted the nanotechnology as a promise alternative in the current context of Brazilian technological innovation involving animal health, as well as a possible diagnostic tool and highlighting its potential therapeutic use in disease control in veterinary medicine. Regarding future perspectives, we believed that greater investment in science and technology could contribute to the advancement and strengthening of nanotechnology in Brazil.

Considering the therapeutic potential of medicinal plants as alternatives to antibiotic therapy, the research aims to evaluate the antimicrobial activity of Commiphora leptophloeos against isolates of Staphylococcus spp. from the milk of ruminants with subclinical mastitis. For this, the crude ethanolic extract from the bark and leaves of Commiphora leptophloeos was prepared, with these being chemically characterized by HPLC-DAD-MS and by MALDI-TOF. The extracts were then evaluated as to their antimicrobial effects against 60 isolates of Staphylococcus spp. through the broth microdilution technique to determine the minimum bactericidal concentration. In addition, the extracts were evaluated as to their ability to interfere with biofilm formation and with the already established biofilm. Although all tested extracts showed antimicrobial action, lower MBC values were recorded for the bark extract in the concentration 781.2 μg/mL (25/60). The extracts of the bark and leaves were able to interfere with the initial stages of biofilm formation, but there was no interference of the extract on the established biofilm. There was a high sensitivity of Staphylococcus spp. isolates from subclinical mastitis cases in ruminants when subjected to the extracts from bark and leaves of Commiphora leptophloeos, as well as regarding the ability of extracts to interfere in biofilm formation, indicating their potential in the use for ruminant mastitis therapy. Keywords: Mastitis, Staphylococcus spp., imburana de cambão, phytotherapy, biofilm.

This study aimed to evaluate the in vitro efficacy of commercial disinfectants on Staphylococcus spp., isolated from mastitis cases in goats, and to associate the observed resistance with the presence of the icaD gene. Broth microdilution was employed to evaluate the in vitro antimicrobial activity of the disinfectants, whereas the Congo Red technique was used for the evaluation of biofilm production and amplification of the icaD gene. All evaluated samples were sensitive to disinfectants, with the following ranges of activity: quaternary ammonium (0.13 – 21.33 µg/ml), chlorhexidine (4.00 – 313.00 µg/ml) and iodine (190.00 – 12500.00 µg/ml); however, the sodium hypochlorite-based disinfectant showed no bactericidal activity in the concentration range from 15.0 to 0.03 μg/ml. The icaD gene presented a frequency of 14.7% in the isolate samples. Fisher's exact test showed a significant effect of the relation between the minimum bactericidal concentration value of the quaternary ammonium-based disinfectant and the presence/absence of the icaD gene (P

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.

Abstract The development of polypyrrole-based nanocomposites as alternative antibacterial agents represents a promising strategy to be applied against the prevailing multi-resistant bacteria. Herein, it is reported the most recent development of antibacterial materials based on the combination of polypyrrole and different fillers (metal nanoparticles, carbon nanotubes, and polysaccharides) and strategies to improve their action (such as light and electrical stimulus). The synergistic interaction of electrostatic forces provided by charged polypyrrole combined with the permeation of nanoparticles through the cell wall favors the leakage of cytoplasmic components and reinforces the antibacterial activity of the resulting material, observed in all-organic composites of polypyrrole and chitosan that reached superior performance against Escherichia coli (108 CFU) or metal-polymer composites (polypyrrole-palladium) with an outstanding performance against different types of bacteria. The development of binary and ternary composites is explored to potentialize the antibacterial synergy of components.

Antibiotic resistance is one of the biggest health challenges of our time. We are now facing a post-antibiotic era in which microbial infections, currently treatable, could become fatal. In this scenario, antimicrobial peptides such as bacteriocins represent an alternative solution to traditional antibiotics because they are produced by many organisms and can inhibit bacteria, fungi, and/or viruses. Herein, we assessed the antimicrobial activity and biotechnological potential of 54 Streptococcus agalactiae strains isolated from bovine mastitis. Deferred plate antagonism assays revealed an inhibition spectrum focused on species of the genus Streptococcus—namely, S. pyogenes, S. agalactiae, S. porcinus, and S. uberis. Three genomes were successfully sequenced, allowing for their taxonomic confirmation via a multilocus sequence analysis (MLSA). Virulence potential and antibiotic resistance assessments showed that strain LGMAI_St_08 is slightly more pathogenic than the others. Moreover, the mreA gene was identified in the three strains. This gene is associated with resistance against erythromycin, azithromycin, and spiramycin. Assessments for secondary metabolites and antimicrobial peptides detected the bacteriocin zoocin A. Finally, comparative genomics evidenced high similarity among the genomes, with more significant similarity between the LGMAI_St_11 and LGMAI_St_14 strains. Thus, the current study shows promising antimicrobial and biotechnological potential for the Streptococcus agalactiae strains.

Abstract Some species of Lactobacillus have demonstrated beneficial health effects being applied in the production of food supplements. Thus, the incorporation of viable cells as encapsulated probiotics represents an essential condition to be considered in new strategies for the controlled release of microorganisms. Herein, the massive encapsulation of Lactobacillus paracasei is provided by the use of alternative electrospinning technique. Is spite of the high voltage required for the production of fibers, a high density of viable cells is observed into the polymeric electrospun web, allowing the controlled release at targeted pH (characteristic of Eudragit® L100 polymer support). The reported procedure circumvents typical drawbacks of degradation of microorganisms under adverse conditions (storage, package and low pH) and preserves its biologic action after complete release from polymer fibers.

Goat and sheep farming in Brazil relies on an expressive herd, especially in the Northeast region of the country, where small ruminants constitute a relevant economic activity, with milk production being an important source of income in some dry areas of the Northeastern states. However, there are still some sanitary problems in the management of dairy goats, among which mastitis is highlighted, especially that of the subclinical type. Therefore, the aim of this study was to evaluate the use of antibiotics, exclusively or in combination with vaccines, in the control of sub-clinical mastitis in small ruminants. The study was conducted on a rural property located in Santa Maria da Boa Vista-PE. Fifty-three female Saanen dairy goats were allocated into four experimental groups. An antibiotic therapy with a commercial antimicrobial based on cloxacillin was used in group 1. Group 2 received the commercial vaccine indicated for herds with an incidence of Staphylococcus strains and the same antibiotic described for Group 1. Group 3 was evaluated for the exclusive use of vaccination, and Group 4 was the control group. A microbiological examination of the milk was carried out to confirm cases of mastitis. The percentage of udders, infected by coagulase-negative Staphylococcus spp. (CNS) before and after the antibiotic + vaccine treatment was 42.8 and 7.1%, respectively. It was observed that the cloxacillin-based antibiotic could be used alone or in combination with mastitis vaccines to control staphylococcal mastitis in small ruminants. Vaccination alone was not effective in reducing the number of mastitis cases; therefore, a cross reaction against CNS was not observed. Thus, further studies to determine the potential of vaccines to control sub-clinical mastitis in small ruminants are necessary.

Mastitis is responsible for both damage to animal health and economic losses to the industry. To identify effective treatments for this disease, products extracted from a variety of plants with antimicrobial potential have gained attention. The present study aimed to assess the in vitro antibacterial potential of the ethanolic extract of two plant species from the Caatinga biome against bacteria isolated from small ruminants with subclinical mastitis. The leaves of Mimosa tenuiflora and Commiphora leptophloeos were dried and processed to obtain crude ethanolic extracts and their phenolic composition was evaluated. In total, 33 Staphylococcus spp. isolates from the bacterial collection of the Laboratory of Food Quality Control of IF SERTÃO-PE were used for evaluation of biofilm production. Furthermore, an antimicrobial susceptibility test was conducted using the minimal bactericidal concentration (MBC) method against the two ethanolic extracts. The toxic potential was measured through a toxicity test with Artemia salina. The quantification of the phenolic compounds revealed that the ethanolic extracts of M. tenuiflora and C. leptophloeos possessed higher amounts of myricetin (43.2 and 294.9 mg in 10 g, respectively) in relation to the other compounds. A 39.4% positivity rate was observed in the nuc gene investigation. The biofilm production analysis revealed that 96.9% of the isolates produced biofilm, evidencing the evolution the microorganisms regarding the development of resistance mechanisms. The MBC results showed an inhibition range between 195.30 and 3125.00 µg mL-1 and between 781.20 and 6250.00 µg mL-1 for the extracts of M. tenuiflora and C. leptophloeos, respectively. The M. tenuiflora extract showed the highest activity, suppressing 100% of the bacterial isolates (n=26), whereas the extract of C. leptophloeos showed an inhibition percentage of 69.23%. The crude ethanolic extract (EEB) of M. tenuiflora was found to be toxic, presenting a DL50 of 118.356 µg mL-1. In contrast, the EEB of C. leptophloeos was found to be non-toxic (DL50 = 1527.430 µg mL-1). In conclusion, both native Caatinga species presented antibacterial activity and myricetin was the major compound. These findings highlight the need for further studies regarding the identification of anti-mastitis products from natural extracts.

A linfadenite caseosa é uma enfermidade que acomete os caprinos e ovinos causando prejuízos econômicos. É uma doença infectocontagiosa de ocorrência mundial, principalmente nos países em desenvolvimento, com ocorrência em todo rebanho brasileiro, especialmente nos Estados do Nordeste, notadamente na Bahia e em Pernambuco. Apesar de ter uma alta prevalência, a prevenção, o diagnóstico e o tratamento são negligenciados, aumentando os riscos de contaminação entre os rebanhos e consequentemente a diminuição da produtividade destes animais e a qualidade do produto ao consumidor. Alguns métodos de prevenção estão sendo desenvolvidos, utilizando bactérias mortas, vivas ou inativadas, podendo usar ou não a tecnologia do DNA recombinante. A virulência de Corynebacterium pseudotuberculosis, está associada com os lipídeos da parede celular e a fosfolipase D, bem como a produção de biofilmes. A produção desses genes de virulência aumenta a sobrevivência e a multiplicação do patógeno. O objetivo do estudo foi abordar nessa revisão os impactos econômicos provocados pela Linfadenite Caseosa, epidemiologia, patogenia e fatores de virulência, bem como sinalizar informações sobre diferentes aspectos entre cepas de C. pseudotuberculosis produtoras e não produtoras de biofilme, buscando fornecer subsídios que possam contribuir para o aprimoramento de vacinas e diagnóstico desta enfermidade dos pequenos ruminantes.

The aim of this study was to evaluate the morphogenic, structural, and productive characteristics of Cenchrus ciliaris L. in response to different doses of biofertilizers that originate from the anaerobic biodigestion of goat and sheep fecal matter on Yellow Argisoil. The experiment was performed in a greenhouse, in a completely randomized factorial design using two types of biofertilizers and five dosages (0%, 2.5%, 5.0%, 7.5%, and 10% of soil volume), with four repeats, for the duration of two vegetative cycles. In the first cycle, there was an increase in leaf lifespan and leaf/stem ratio with the use of 10% and 7.5% caprine biofertilizer, respectively. From the second cycle, doses of biofertilizers between 7.5% and 10% led to an improvement in the development of the morphogenic variables - leaf sprouting rate, leaf and pseudostem elongation rate, leaf lifespan, and senescence, as well as the structural characteristics - expanded leaf length and width, number of sprouting, expanded and live leaves, and expanded/expanding leaf ratio, resulting in a 73% increase in biomass. It was concluded that the use of biofertilizers improves the main morphogenic and structural characteristics of Cenchrus ciliaris L., resulting in increased fodder mass yield.

ABSTRACT: This study aimed to perform phytochemical analysis and to test the antimicrobial activity of the crude hydroalcoholic extract obtained from the leaves of Sphagneticola trilobata. Classes of secondary metabolites present in the extract were identified through phytochemical screening using analytical thin-layer chromatography. Antimicrobial activity was evaluated by testing cultures of Staphylococcus aureus, S. epidermidis, Staphylococcus spp., Escherichia coli, Serratia marcescens, Enterococcus faecalis, Pseudomonas aeruginosa, Salmonella Typhimurium, and Klebsiella pneumoniae isolated from human skin and those of Staphylococcus spp. isolated from dog skin using the broth microdilution method. In the phytochemical screening, classes of anthracenic derivatives and mono-, sesqui-, and diterpenes were identified. Colorimetric analysis showed total phenol and total flavonoid contents of 21.7 ± 0.009 mg of gallic acid equivalents per gram of sample and 0.23 ± 0.005 mg of catechin equivalents per gram of sample, respectively. Microbiological analysis revealed that the hydroalcoholic extract of S. trilobata exhibited antimicrobial activity against cultures of Staphylococcus spp., E. coli, S. marcescens, and E. faecalis isolated from human skin and those of Staphylococcus spp. isolated from dog skin. Thus, crude hydroalcoholic extract of leaves of S. trilobata contained flavonoids and terpenoids as secondary metabolites, which contributed to its antimicrobial activity against skin bacteria isolated from different sources.

ABSTRACT: Prototheca spp. have been reported as an emergent environmental mastitis pathogen in several countries. Biofilm formation is a significant factor associated with different degrees of virulence developed by many microorganisms, including Prototheca spp. The present study aimed to compare two growth conditions and two staining dyes to determine which combination was more appropriate to evaluate qualitatively and quantitatively the production of biofilm by P. zopfii. Biofilm formation was evaluated in polystyrene microplates under static and dynamic growth conditions and staining with crystal violet or cotton blue dye. All P. zopfii isolates from cows with mastitis were classified as biofilm-producers in all growth conditions and staining. The cotton blue dye proved to be more appropriate method to classify the intensity of P. zopfii biofilm production.

This study aimed to evaluate the chemical composition of milk after infection and treatment of mastitis caused by Staphylococcus aureus. Ten Saanen goats were used. Experimental infection was performed in 20 mammary glands, forming four groups, each consisting of five mammary glands. One group was treated with commercial antimicrobial and another with natural plant extract-based ointment. The other groups were constituted as control. The values obtained for fat, lactose, protein, total solids, and milk production were analyzed through the experimental moments, comparing them among each other and between experimental groups. Mastitis was diagnosed by milk microbiology after 24 hours of inoculation. The animals showed no changes in the clinical parameters evaluated. For the percentage of fat, protein, and total solids, there was an increase over the experimental moments. However, there was variation among the experimental groups. Given the importance of conventional antimicrobial therapy and the increased use of natural antimicrobials, obtained results showed equality between the two treatment options, especially regarding the maintenance of the main milk components. Keywords: goat; milk composition; therapy; mastitis.

ABSTRACT: The aim of the present study was to assess two diagnostic techniques (California mastitis test (CMT) and the somatic cell count (SCC)) that can diagnose mastitis in dairy goats. Experimental infection was conducted using 20 mammary glands, a strain of Staphylococcus aureus, an infectious dose of 1.2x108CFU mL-1 and a volume of 1mL per mammary gland. The CMT and the SCC were used to detect subclinical mastitis. Bacterial culture (BC) was performed immediately after milk collection and was used as the gold standard. Four experimental time points were established (0, 24, 48 and 72 hours post-inoculation). Analysis of the ROC curve confirmed that the best combination of sensitivity and specificity were obtained with a cutoff point of 405.5, 6030.0 and 729.5x103 cells mL-1, respectively at M1, M2 and M3. Furthermore, considering the drop in sensitivity throughout the experimental time points, the use of serial bacterial cultures are recommended, particularly in herds with a high prevalence of S. aureus.

Loss in the supply chain of small ruminants owing to condemnations of carcasses in the abattoirs and slaughterhouses is common in northeastern Brazil. This study aims to identify bacterial agents, including Mycobacterium spp., in the abscesses found in the postmortem analysis of the carcasses of sheep and goats bred in northeastern Brazil. Our analysis involved 679 goats and 1,838 sheep carcasses. Abscess samples were extracted and inoculated on blood agar and Lowenstein Jensen with pyruvate or glycerol for bacterial isolation. We then performed polymerase chain reaction of the hps 65 gene; samples positive for Mycobacterium spp. were subjected to DNA sequencing. Relative frequencies of abscesses in goats and sheep were 5.44 and 3.26%, respectively. Microbiological analysis revealed 87.7% bacterial growth in the inoculated samples. Among these, Corynebacterium pseudotuberculosis represented 67.7% of the isolates. We observed 1.9% mycobacteria growth in the abscess samples inoculated on Lowenstein-Jensen medium. PCR of DNA extracted from abscesses samples showed amplification of 0.9% of samples. After sequencing, Mycobacterium spp. isolate was identified as M. novocastrense. C. pseudotuberculosis was the main agent responsible for the formation of abscesses in the examined animals, and we did not identify any species of the M. tuberculosis complex in the examined small ruminants.

The aim of the present study is to assess the antibacterial potential of plants from the Caatinga biome of the semi-arid region of Pernambuco, against Staphylococcus spp. isolates from cases of subclinical mastitis in small ruminants, such as goats and ewes. Ethanolic extracts of the following plants from the Caatinga biome were used: Encholirium spectabile Mart., Bromelia laciniosa Mart., Neoglaziovia variegata Mez., Amburana cearensis (Fr. Allem.) A.C.Smith, Hymenaea martiana Hayne, and Selaginella convoluta Spring. The presence of phenolic components, flavonoids, steroids, and terpenoids was verified for all the tested extracts. The mean minimal bactericide concentrations of the extracts of E. spectabile, B. laciniosa, and N. variegate were 11,379, 11,405, and 11,995 μg/mL, respectively. The highest inhibitory activities were observed for A. cearensis and H. martiana, which inhibited 88.1 and 99.4% of the isolates, respectively. Other studies focusing on in vitro and in vivo activities should be undertaken.

Goat and sheep production in the semi-arid northeast of Brazil has shown great economic potential. However, health problems can compromise the productivity of these animals. Given the scarcity of studies about the occurrence of these diseases, the aim of the present study was to analyze the serological diagnosis of anti-Brucella and anti-lentivirus antibodies among small ruminants in municipalities located in the Brazilian states of Bahia and Pernambuco. The samples were collected from local slaughterhouses and dairy farms. In total, 997 serum samples from animals in slaughterhouses and dairy herds were collected. In order to diagnose the caprine arthritis encephalitis virus (CAEV), the samples underwent agarose gel immunodiffusion (AGID) testing. The buffered acidified antigen test (goats) and agarose gel immunodiffusion test (sheep) were used to detect anti-Brucella abortus and B. ovis antibodies following the methodology recommended by the Institute of Technology of Paraná (TECPAR). With anti-CAEV antibodies, seropositivity rates of 4.1% and 2.2% were recorded for animals from the slaughterhouses and dairy farms, respectively. None of the animals (goats or sheep) were positive for anti-B. abortus antibodies. With B. ovis, a seropositivity rate of 6.5% (n = 13) was recorded among the 199 sheep serum samples. Results of the present study confirmed the presence of the CAE virus in the meat and dairy herds studied, although the prevalence was low. Natural infection by B. abortus did not occur in the goat and sheep herds assessed. Seropositivity for B. ovis was confirmed, although prevalence was low. Direct tests are required to diagnose ovine brucellosis.

ABSTRACT: Aeromonas hydrophila is a common fish pathogen that causes extensive damage to aquaculture. To develop and implement a more adequate strategy to farm fish, it is crucial to understand the bacterial-resistance levels and their transference dynamics. The objective of this study was to analyze the resistance profile of isolated Aeromonas hydrophila to antimicrobial agents and heavy metals and draw a correlation of the observed profiles with the presence of plasmids. Resistance of the isolated bacteria to antimicrobial agents (oxacilin, gentamicin, tetracycline, and nalidixic acid) and heavy metals (cadmium, lead, copper, and manganese) was verified using the minimum bactericidal concentration (MBC) and minimum inhibitory concentration (MIC) standards. The Multiple Antibiotic Resistance Index (MAR Index) was calculated. Plasmids were extracted by using a common methodology described elsewhere. Mann-Whitney Test, implemented in the R environment, was used to determine the correlation between resistance and plasmids presence. A high resistance to almost all antimicrobial agents and heavy metals was observed, except to gentamicin and cadmium. The MAR index results showed resistance to all antimicrobial profiles. Of the isolated bacteria, 14 showed the presence of plasmids. However, no correlation was noted between the resistance profile and the plasmid presence for these isolates, indicating that the genes responsible for resistance to microbial agents and heavy metals are present in the cromossomic DNA, which in turn suggested the possibility of gene transfer between the isolated bacteria. The resistance to heavy metals can be linked to heavy utilization of fertilizers along the Sao Francisco River.

Neonatal diarrhea determines significant changes in feed conversion, causing productivity loss in caprine herds. The antimicrobial resistance in bacteria is characterized as an important public health issue; therefore, Escherichia coli may be characterized as an important pathogen due to expressing virulence mechanisms responsible for significant clinical conditions in humans and animals. The present study evaluated the presence of E. coli among 117 caprine fecal samples and analyzed the isolates for antimicrobial resistance. Suggestive colonies were submitted to biochemical screening followed by genotypic group determination and phylogenetic analysis; further, the samples were submitted to antimicrobials susceptibility test. E. coli, Salmonella spp, Shigella sonnei and Enterobacter aerogenes were identified. E. coli isolates were phylogenetically classified as B2 (9/39), D (19/39), B1 (7/39) e A (4/29) groups. The analysis of the isolates also revealed the presence of K99 (04/39) and Stx (02/39) virulence factors. Antimicrobial susceptibility test revealed sensitive isolates to Chloramphenicol, Streptomycin, Amoxicillin and Ciprofloxacin, being all resistant to Lincomycin, Vancomycin and Penicillin. The results support the need of establishing restricted protocols for antimicrobial use, a fundamental procedure for health improvement in Brazilian caprine herds.

We evaluated the physical and chemical characteristics of raw milk from goats supplemented with levels of Total Digestible Nutrients (TDN) in the dry period. Samples from 20 goats were divided into 4 groups, 3 groups received 400g concentrate per animal with different levels of TDN (65, 75 and 85%), and the control group (without concentrate). Four samples of milk were used for each animal, in four sampling periods, which were subjected to physical and chemical analysis. There was no significant difference between the treatments for the parameters of acidity and fat. The diet without concentrate presented higher value for the cryoscopic index. Mean values of the four treatments were within the range set by the Brazilian legislation for all parameters evaluated. The use of concentrate, containing 85% TDN in goat supplementation, was favored the physical and chemical characteristics of the milk during the dry period.

Reproductive tract infections are the main causes of losses from the low reproductive efficiency of sheep. Gram negative bacilli belonging to the normal flora of the genital region can trigger diseases. The pathogenicity of these agents is expressed when females are with weakened immune system, either by food or stress management. Flaws in and concern about antibiotic residues in animal production have prompted research regarding alternatives for the treatment of diseases. The herbal medicine hás been considered in this context is the subject of numerous studies. This study aimed to evaluate the antibacterial potential of ethanol extracts of plants belonging to the flora of the Northeast against gram negative bacilli isolated from cervical-vaginal mucosa of sheep. Six plants were selected from Caatinga biome: Encholirium spectabile, Bromelia laciniosa, Neoglaziovia variegata, Amburana cearensis Hymenaea martiana and Selaginella convoluta. The plant material was processed to obtain the crude extract. This was tested by microdilution plate and determining the minimum bactericidal concentration, the second document of Clinical and Laboratory Standards Institute (CLSI) and the extracts diluted in water and alcohol. We used 43 gram negative isolates, as follows: 14 E. coli, 10 Enterobacter spp., 10 Acinetobacter spp. 9 and Klebsiella spp. In the aqueous dilution Klebsiella spp. showed response only to species B. laciniosa, S. convoluta and H. martiana. All tested extracts showed antibacterial activity against Acinetobacter spp and no activity against E. coli and Enterobacter spp. Among the extracts diluted in water H. martiana showed the highest antibacterial activity. In all dilution alcoholic extracts showed inhibitory activity against all bacterial genera, but no statistical difference between them.

O presente estudo avaliou o efeito de diferentes processamentos de ração e da alimentação com alimento vivo sobre o desempenho e sobrevivência de alevinos de pacamã (Lophiosilurus alexandri). Foram utilizados 140 alevinos de pacamã distribuídos em um delineamento inteiramente casualizado, com quatro tratamentos e cinco repetições, durante 30 dias. Os tratamentos consistiram na alimentação dos alevinos com pós-larvas de tilápia do Nilo e com rações nas formas farelada, micropeletizada e úmida. Houve um melhor desempenho (P0,05) sobre a sobrevivência dos peixes. A alimentação viva proporciona melhor desempenho para alevinos de pacamã comparada à alimentação inerte.

In aquaculture, infections caused by bacteria are factors that cause damage. The purpose of this study was to identify bacteria with pathogenic potential in pacamã (Lophiosilurus alexandri) of São Francisco Valley. One hundred and fifteen animals were used for sampling from gills, kidneys, external lesions and eggs. The samples were streaked in Trypticase soy agar. The biochemical and antimicrobial susceptibility profiles were determined. The bacterial species identified were A.hydrophila, A. salmonicida, A. media, Acinetobacter spp., P. alcaligenes, E. aerogenes, E. agglomerans, K. oxytoca, K. pneumoniae, P.shigelloides, V. parahaemo- lyticus, V. metschnikovii and S. sannei. The sensitivity to antimicrobial was 85% to norfloxacin, 79% to ceftri- axone, 78% to enrofloxacin, 68% to sulfazothrim, 60% to nitrofurantoin, 59% to tetracycline, 55% to nalidixic acid, 49% to streptomycin, 45% to erythromycin, 32% to neomycin, 7% to ampicillin and 3% to lincomycin. Multiple resistance was observed to all isolates analyzed. Considering the occurrence of pathogenic bacteria in pacamã and its resistance to antimicrobial drugs, security measures should be adopted during antimicrobialther- apy, with the use of antibiotics that did not show resistance strains or trying natural products as an alternative to antibiotics, ensuring protection of human and animal health, as well as the environment.

O objetivo deste trabalho foi quantificar os teores de fenóis e flavonoides totais, bem como avaliar as atividades antioxidante e antimicrobiana de extratos obtidos dos talos e folhas de atemoia (A. cherimola Mill. x A. squamosa L.), que pertence à família Annonaceae. A atividade antioxidante foi avaliada pelos métodos de sequestro dos radicais 2,2-difenil-1-picrilhidrazil (DPPH) e 2,2'-azinobis-3-etilbenzotiazolina-6-ácido sulfônico (ABTS), bem como pelo método da cooxidação do β-caroteno/ácido linoleico. A avaliação da atividade antimicrobiana dos extratos foi analisada contra 10 cepas de bactérias. Os resultados da atividade antioxidante dos extratos mostraram que o extrato etanólico dos talos (EEt) foi o antioxidante mais efetivo (IC50 = 10,44 ± 1,25 µg/mL) no método do sequestro do DPPH, bem como no sequestro do radical ABTS (24,81 ± 0,49%). O extrato hexânico das folhas apresentou o melhor percentual de atividade antioxidante no ensaio do β-caroteno/ácido linoleico (41,12 ± 4,35%). Os extratos etanólico dos talos e metanólico das folhas mostraram-se ativos contra cepas de Bacillus cereus, Klebsiella pneumoniae, Staphylococcus aureus resistente à meticilina (MRSA), Staphylococcus aureus e Staphylococcus epidermidis.

Objetivou-se com esse estudo avaliar a sensibilidade da PCR e PCR nested na detecção de T. gondiino sangue de ovelhas experimentalmente infectadas com taquizoítos via sêmen. Foram colhidas amostras de sangue de 30 ovelhas infectadas com diferentes doses: G1- 6,5 X 104 taquizoítos; G2- 4 X 107 taquizoítos e G3- grupo controle. Para a pesquisa de anticorpos anti-T. gondii foi utilizada a técnica de Imunofluorescência Indireta (IFI). Para o PCR e PCR nested foram utilizados iniciadores derivados do gene B1. Observou-se soroconversão apenas dos grupos infectados. Na PCR, detectou-se o DNA do T. gondii em 40% e na PCR nested em 93,3% amostras de sangue das ovelhas infectadas. No G3 não foi detectado anticorpos e DNA parasitário em nenhum animal.Conclui-se que a PCR nested é mais sensível na detecção de T. gondii no sangue de animais experimentalmente infectados via sêmen, mostrando resultado superior ao da PCR e podendo ser utilizada com êxito em estudos dessa natureza.

Rhodococcus equi é um micro-organismo intracelular facultativo, agente etiológico da rodococose, uma importante enfermidade que acomete principalmente potros com menos de seis meses de idade, causando a morte geralmente em decorrência de lesões pulmonares. Este agente também tem potencial zoonótico e emergiu como um patógeno oportunista no mundo, acometendo humanos imunocomprometidos, especialmente os transplantados e infectados pelo vírus da imunodeficiência humana (HIV). Entretanto, infecções por R. equi em hospedeiros hígidos tem sido relatadas, principalmente em crianças e idosos. Estudos tem mostrado um nível crescente na resistência de isolados de R. equi em relação aos antimicrobianos comumente utilizados no tratamento de animais e seres humanos infectados por este agente. A virulência deste pode estar associada a fatores como a cápsula de polissacarídeo, fosfolipase C e à enzima colesterol oxidase (fator equi). No entanto, uma proteína localizada em um plasmídeo, designada vapA, é essencial para a sobrevivência e replicação do agente em macrófagos. Com isso, os objetivos deste estudo foram avaliar o perfil de suscetibilidade de isolados de R. equi de diferentes fontes em relação aos antimicrobianos mais comumente utilizados na terapêutica animal e humana, bem como verificar a associação entre a presença do gene vapA e o índice de resistência múltipla aos antimicrobianos (IRMA). Neste estudo, 67 isolados brasileiros de R. qui de diferentes fontes foram analisados: 30 provenientes de amostras clínicas de equinos, sete de humanos e 30 ambientais (seis do solo e 24 de fezes de equinos). Para avaliar o perfil de suscetibilidade dos isolados utilizou-se o método de disco difusão, sendo testadas 16 drogas de diferentes classes de antimicrobianos. As amostras clínicas de equinos apresentaram as maiores taxas de resistência à penicilina (86,7%) e lincomicina (30%). Além disso, foram também resistentes a macrolídeos (azitromicina a 6,7%, eritromicina a 6% e claritromicina a 3,3%) e rifamicina (13%). Todas as amostras humanas e ambientais foram sensíveis aos macrolídeos e rifamicina. Contudo, isolados ambientais demonstraram níveis elevados de resistência à penicilina e cloranfenicol. Da mesma forma, os isolados humanos apresentaram alto nível de resistência ao ceftiofur, lincomicina e sulfazotrim. O IRMA em todos os isolados de R. equi variou de 0 a 0,67, tendo como valores médios 0,19 para as amostras clínicas de equinos, 0,14 nas ambientais e em isolados humanos foi de 0,1. Apesar da alta sensibilidade observada nos isolados analisados, verificaram-se diferentes níveis de resistência nas amostras clínicas de equinos. Em contraste, os isolados ambientais não demonstraram resistência em relação aos agentes antimicrobianos utilizados na terapia da rodococose equina. Além disso, em isolados humanos não se observou resistência contra a droga para uso restrito em terapia de humano. Com base no IRMA observado em isolados clínicos de equinos, destacamos a importância de medidas restritivas e mais cautela na utilização de antimicrobianos em infecções causadas por R. equi para evitar o aumento de novas cepas multirresistentes.

Corynebacterium diphtheriae, Corynebacterium ulcerans and Corynebacterium pseudotuberculosis constitute a group of potentially toxigenic microorganisms that are related to different infectious processes in animal and human hosts. Currently, there is a lack of information on the prevalence of disease caused by these pathogens, which is partially due to a reduction in the frequency of routine laboratory testing. In this study, a multiplex polymerase chain reaction (mPCR) assay that can simultaneously identify and determine the toxigenicity of these corynebacterial species with zoonotic potential was developed. This assay uses five primer pairs targeting the following genes: rpoB (Corynebacterium spp), 16S rRNA (C. ulcerans and C. pseudotuberculosis), pld (C. pseudotuberculosis), dtxR (C. diphtheriae) and tox [diphtheria toxin (DT) ]. In addition to describing this assay, we review the literature regarding the diseases caused by these pathogens. Of the 213 coryneform strains tested, the mPCR results for all toxigenic and non-toxigenic strains of C . diphtheriae, C. ulcerans and C. pseudotuberculosis were in 100% agreement with the results of standard biochemical tests and PCR-DT. As an alternative to conventional methods, due to its advantages of specificity and speed, the mPCR assay used in this study may successfully be applied for the diagnosis of human and/or animal diseases caused by potentially toxigenic corynebacterial species.

This study aimed to determine the antimicrobial susceptibility profile and to assess the presence of mechanisms of antimicrobial resistance in Staphylococcus spp. (n=36) isolated from mastitis in sheep’s Chapecó-SC. The potential for biofilm production was determined by phenotypic tests of Congo Red Agar, DAPI and Gentian Violet and by PCR for the detection of icaD gene. To evaluate the antimicrobial resistance testing was performed disk diffusion and detection of resistance genes blaZ, mecA, ermA, ermB and ermC and msrA also performed by PCR.The pump test was conducted by efuxo crop growth Muller Hinton agar containing ethidium bromide. The results showed that 1 (2,78%), 36 (100%) and 10 (27,78%) isolates were considered to produce a biofilm on Congo Red Agar test, Gentian Violet and DAPI, respectively, while the gene icaD was observed in only 2 (5.55%) isolates. The lowest percentage of sensitivity was observed for ampicillin (58.33%) and penicillin (58.33%). All strains tested were negative for the mecA, ermA, ermB and ermC genes. However, the isolates were positive for other resistance genes, being the blaZ and the msrA, with percentages of positivity of 58.33% and 11.11% respectively. Only one sample was positive for efflux pump test.O presente trabalho teve como objetivos determinar o perfil de sensibilidade aos antimicrobianos e avaliar a presença de mecanismos de resistência antimicrobiana em Staphylococcus spp. (n=36) isolados de mastite em ovelhas do município de Chapecó-SC. O potencial para produção de biofilme foi determinado pelos testes fenotípicos de Agar Vermelho Congo, DAPI e Violeta de Genciana e por teste molecular pela técnica de PCR para a detecção do gene icaD. Para determinar o perfil de resistência aos antimicrobianos, foi realizado o teste de difusão em disco e detecção dos genes de resistência blaZ, mecA, erm (A, B e C) e msrA. O teste da bomba de efuxo foi realizado através do crescimento das culturas em Agar Muller Hinton contendo brometo de etídio. Os resultados mostraram que 1 (2,78%), 36 (100%) e 10 (27,78%) isolados foram considerados produtores de biofilme pelos testes de Agar Vermelho Congo, DAPI e Violeta de Genciana, respectivamente, enquanto que o gene icaD foi observado em apenas 2 (5,55%) isolados. O menor percentual de sensibilidade foi observado para ampicilina (58,33%) e penicilina (58,33%). Os isolados avaliados foram positivos para os genes de resistência blaZ (58,33%) e msrA (11,11%). Nenhum isolado apresentou os genes de resistência erm (A, B, C) e apenas uma amostra foi positiva para o teste da bomba de efluxo.

Para verificar a dinâmica da resistência aos antimicrobianos em uma propriedade rural no município de Santa Maria da Boa Vista, PE, foram avaliados 14 isolados de Staphylococcus epidermidis de caprinos com mastite subclínica. O perfil de resistência aos antimicrobianos foi determinado pelo teste de difusão em disco. A genotipificação foi realizada empregando o marcador REP (Repetitive Extragenic Palindromic) - PCR, utilizando o primer RW3A, enquanto os graus de similaridade e o fenograma de agrupamento foram estabelecidos por meio do coeficiente de Sorensen-Dice (SD) do algoritmo UPGMA, programa NTSYS-pc, o qual permitiu a identificação de 4 padrões dos 14 isolados de S. epidermidis, sendo oito no perfil A, quatro no perfil B, um no perfil C e um no perfil D. Para todos os grupos, a resistência à penicilina foi observada, enquanto que, para os grupos A e C, esta foi associada à lincomicina, no grupo B, esta foi associada à tetraciclina.To verify the dynamics of antimicrobial resistance in a property in Santa Maria da Boa Vista city - Pernambuco were evaluated 14 Staphylococcus epidermidis isolates from goats with subclinical mastitis. The profile of antimicrobial resistance was determined by disk diffusion test. The genotyping was performed using the marker REP (Repetitive Extragenic Palindromic) - PCR, using RW3A primer, where the degrees of similarity and clustering phenogram were established by means of the Sorensen-Dice coefficient (SD) algorithm UPGMA, program NTSYS-pc, which allowed the identification of 4 patterns of 14 S. epidermidis isolates, being eight in the profile A, four in a profile B, one in profile C and one in profile D. For all groups to penicillin resistance was observed, while for groups A and C this was associated with lincomycin, for group B this was associated with tetracycline.

BACKGROUND: Laser therapy is a low cost, non-invasive procedure with good healing results. Doubts exist as to whether laser therapy action on microorganisms can justify research aimed at investigating its possible effects on bacteria-infected wounds. OBJECTIVE: To assess the effect of low intensity laser on the rate of bacterial contamination in infected wounds in the skin of rats. METHODS: An experimental study using 56 male Wistar rats. The animals were randomly divided into eight groups of seven each. Those in the "infected" groups were infected by Staphylococcus aureus MRSA in the dorsal region. Red laser diode (AlGaInP) 658nm, 5J/cm2 was used to treat the animals in the "treated" groups in scan for 3 consecutive days. Samples were drawn before inoculating bacteria and following laser treatment. For statistical analysis we used the nonparametric Wilcoxon (paired data) method with a significance level of p RESULTS: The statistical analysis of median values showed that the groups submitted to laser treatment had low bacterial proliferation. CONCLUSION: The laser (AlGaInP), with a dose of 5J/cm2 in both intact skin and in wounds of rats infected with Staphylococcus aureus MRSA, is shown to reduce bacterial proliferation.FUNDAMENTOS: Fundamentos: A terapia a laser é um procedimento de baixo custo, não invasiva e com bom desempenho na cicatrização. As dúvidas existentes quanto a sua ação sobre microrganismos justifica a realização de pesquisas visando investigar os possíveis efeitos em feridas infectadas por bactérias. OBJETIVO: Avaliar o efeito do laser de baixa intensidade sobre a taxa de contaminação bacteriana em feridas infectadas na pele de ratos.MÉTODOS:Estudo experimental, utilizando 56 ratos machos Wistar. Os animais foram distribuídos aleatoriamente em oito grupos de sete animais. Nos animais dos grupos lesionados foi realizada uma incisão na região dorsal.Os animais dos grupos infectados foram infectados por Staphylococcus aureus MRSA. os animais dos grupos tratados foram tratados com laser de Diodo vermelho (AlGaInP) 658nm, 5J/cm2 em varredura, durante 3 dias consecutivos. Foi colhida uma amostra antes de inocular as bactérias e outra após o tratamento com laser. Para a análise estatística foram utilizados os testes não paramétricos de Wilcoxon (dados pareados). Considerando como significante pRESULTADOS: Através da análise estatística das medianas, observou-se que os grupos submetidos ao laser apresentavam uma proliferação bacteriana menor. CONCLUSÃO:O laser (AlGaInP), com uma dose de 5J/cm2, tanto em feridas quanto em pele íntegra de ratos infectados por Staphilococcus aureus MRSA, se mostrou capaz de reduzir a proliferação bacteriana.

Caseous lymphadenits (CL) is a chronic and subclinical disease that affects goats and sheep and, consequently, causes economic losses, especially to small producers. The purpose of this study, through use of Polymerase Chain Reaction (PCR), was to verify the presence of virulence genes of phospholipase D (PLD), integral membrane protein (FagA), iron enterobactin transporter (FagB), ATP binding cytoplasmic membrane protein (FagC) and iron siderophore binding protein (FagD) in 168 isolates of C. pseudotuberculosis obtained from cases of caseous lymphadenitis in goats and sheep. FagA, FagB and PLD genes were detected in all 145 strains isolated from abscesses in superficial lymph nodes and in 23 strains isolated from viscera. The FagC gene was positive in 167 (99.40%) isolates. The FagD gene was detected in 160 (95.23%) isolates. All virulence factors analyzed were found more frequently among isolates collected in the viscera of animals with CL, indicating a multifactorial nature, as well as variations, in the invasive potential of C. pseudotuberculosis strains.

Neste estudo, objetivou-se avaliar a composição química do leite após a infecção e tratamento da mastite induzida por S. Aureus. Foram utilizadas 10 fêmeas caprinas da raça Saanen. Fez-se a infecção experimental em 20 glândulas mamárias, formando-se quatro grupos experimentais, sendo cada um composto por cinco glândulas mamárias. Um grupo foi tratado com antimicrobiano comercial e outro com pomada à base de extrato natural de planta. Os demais grupos foram constituídos controles. Os valores obtidos para gordura, lactose, proteína, sólidos totais e produção leiteira foram analisados ao longo dos momentos experimentais, comparando-os entre si e entre os grupos experimentais. A mastite foi diagnosticada 24 horas pós inoculação, utilizando-se a lactocultura. Os animais não apresentaram alterações nos parâmetros clínicos avaliados. Para os percentuais de gordura, proteína e sólidos totais, observou-se uma elevação ao longo dos momentos experimentais, sem variação entre os grupos experimentais. Tendo em vista a importância da terapia antimicrobiana convencional e a crescente utilização de antimicrobianos naturais, os resultados obtidos mostram igualdade entre as duas opções terapêuticas, principalmente no aspecto de manutenção dos principais componentes do leite. Palavras-chave: cabras; composição do leite; terapia; mastite.

The purpose of the present study was to evaluate the antimicrobial activity of propolis extracts diluted in different solvents against bacteria from Staphylococcus genus. The study was performed in the Immunology and Microbiology Laboratory from Universidade Federal do Vale do São Francisco. The propolis extracts were prepared using brown propolis diluted in different solvents such as chloroform, methanol, ethyl acetate and grain alcohol. In order to determine the antimicrobial potential of extracts, agar well diffusion method was used, with controls for each diluent. After that, Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) methods were used. All tests were performed in triplicate. In the agar well diffusion test, the measurements of the inhibition zone for propolis extract were as follows: grain alcohol and propolis (2.88mm), methanol and propolis (2.41mm), chloroform and propolis (2.40mm) and ethyl acetate and propolis (0.83mm). The MBC of propolis extracts in different solvents were 93.75 μg/mL for grain alcohol, 375 μg/mL for chloroform and methanol and 3,000 μg/ml for ethyl acetate. Statistically significant differences were achieved comparing the inhibition zones of propolis diluted in grain alcohol and ethyl acetate (2.88 and 0.83 mm, respectively). Considering the low cost of therapy and the activity of the propolis against caprine mastitis pathogens, other studies regarding in vivo activity and chemical characterization are necessary, in addition to evaluation of the toxicological aspects of propolis extracts.The purpose of the present study was to evaluate the antimicrobial activity of propolis extracts diluted in different solvents against bacteria from Staphylococcus genus. The study was performed in the Immunology and Microbiology Laboratory from Universidade Federal do Vale do São Francisco. The propolis extracts were prepared using brown propolis diluted in different solvents such as chloroform, methanol, ethyl acetate and grain alcohol. In order to determine the antimicrobial potential of extracts, agar well diffusion method was used, with controls for each diluent. After that, Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) methods were used. All tests were performed in triplicate. In the agar well diffusion test, the measurements of the inhibition zone for propolis extract were as follows: grain alcohol and propolis (2.88mm), methanol and propolis (2.41mm), chloroform and propolis (2.40mm) and ethyl acetate and propolis (0.83mm). The MBC of propolis extracts in different solvents were 93.75 μg/mL for grain alcohol, 375 μg/mL for chloroform and methanol and 3,000 μg/ml for ethyl acetate. Statistically significant differences were achieved comparing the inhibition zones of propolis diluted in grain alcohol and ethyl acetate (2.88 and 0.83 mm, respectively). Considering the low cost of therapy and the activity of the propolis against caprine mastitis pathogens, other studies regarding in vivo activity and chemical characterization are necessary, in addition to evaluation of the toxicological aspects of propolis extracts.

Dermatophytosis is one of the main diseases of soft tissue pets, especially dogs and cats, as the etiological agents are responsible for important zoonotic disease sometimes difficult to treat. The presence of pathogenic fungi on the skin lesions associated with alopecia, border demarcated, and itching of mild intensity characterizes dermatophytosis. This study aimed to determine the prevalence of fungal species involved in cases of dermatophytosis in companion animals showing lesions. We performed skin scrapings of skin lesions of 41 (forty-one) dogs and 7 (seven) cats. The samples were sent to the Laboratory of Microbiology UNOESC - Xanxerê for possible isolation. The hair of animals were subjected to direct examination under a microscope to detect the existence of possible changes. Subsequently, it was the cultivation of the samples in Sabouraud agar containing antibiotics. Fungi were identified by macroscopic morphology of the microculture technique. In dogs, the direct examination reported eleven (26.8%) positive by the visualization of lesions and arthroconidia. After cultivation of 41 samples collected from dogs, six (14.6%) showed a growth of dermatophytes. Of the positive samples, 50% (3/6) were identified as Microsporum canis, 33.30% (2/6) and M. gypseum and 16.67% (1/6) and M. nanum. In cats, the seven samples tested by direct examination, three (42.8%) were positive, however no positive cultures were obtained in cats, when the exam. Dermatophytosis was confirmed in a few animals, even upon the occurrence of lesions highly suggestive.A dermatofitose é uma das principais enfermidades tegumentares de animais de companhia, em especial cães e gatos, uma vez que seus agentes etiológicos são responsáveis por importante zoonose por vezes de difícil tratamento. A presença de fungos patogênicos na pele, associado às lesões alopécicas, de bordas delimitadas, e com prurido de discretas intensidades caracteriza a dermatofitose. Este estudo visou determinar a prevalência das espécies fúngicas envolvidas em casos de dermatofitose em animais de companhia apresentando lesões cutâneas. Foram realizados raspados cutâneos de lesões de pele de 41 (quarenta e um) cães e 7 (sete) gatos. As amostras foram remetidas ao Laboratório de Microbiologia da UNOESC – Xanxerê para eventual isolamento. Os pêlos dos animais foram submetidos ao exame direto em microscópio, para detectar a existência de possíveis alterações. Posteriormente, foi realizado o cultivo das amostras em Ágar Saboraud, contendo antibiótico. Os fungos foram identificados através da morfologia dos macroconídeos pela técnica de microcultivo. Nos cães, o exame direto indicou onze (26,8%) amostras positivas pela visualização de lesões e artroconídeos. Após o cultivo, das 41 amostras coletadas de cães, seis (14,6%) evidenciou-se crescimento de dermatófitos. Das amostras positivas, 50% (3/6) foram identificadas como Microsporum canis, 33,30% (2/6) como M. gypseum e 16,67% (1/6) como M. nanum. Nos gatos, das sete amostras submetidas ao exame direto, três (42,8%) foram consideradas positivas, contudo não foram obtidos cultivos positivos nos gatos, quando da realização do exame. A dermatofitose foi confirmada em poucos animais, mesmo quando da ocorrência de lesões bastante sugestivas.

A discutida questão da substituição do uso de antibacterianos em rações (promotores de crescimento) requer urgentes alternativas. Face às necessidades de inibidores microbianos nesses alimentos, os óleos essenciais (OES) se constituem em alternativa, sob avaliação. Neste estudo, avaliou-se a atividade antimicrobiana dos OES de Origanum vulgare (orégano), Thymus vulgaris (tomilho), Cinnamomum zeylanicum (canela), Lippia graveolens (orégano mexicano), Zingiber officinale (gengibre), Salvia officinalis (sálvia), Rosmarinus officinalis (alecrim) e Ocimum basilicum (manjericão) frente a amostras de Escherichia coli isoladas de fezes de aves (n=43) e de bovinos (n=36). A concentração inibitória mínima (CIM) e a concentração bactericida mínima (CBM) foram determinadas para cada isolado através da técnica de microdiluição em caldo, a partir da máxima concentração de 6400µg mL-1 de cada OE testado. Observou-se atividade antimicrobiana para os OES de orégano, orégano mexicano, tomilho, canela. Para todas as amostras testadas, independente de sua origem, os OES mais e menos efetivos quanto à atividade antimicrobiana foram o orégano e a canela, respectivamente. Esses resultados confirmaram o potencial antibacteriano de alguns OES, os quais merecem novas investigações abordando sua adição na alimentação de aves e bovinos.The discussed issue about replacing the use of antibiotics in animal feed (growth promoters) requires emerging alternatives. To meet the needs of microbial inhibitors in these foods, the essentials oils (EOS) constitute potential alternatives under evaluation. In this study it was evaluated the antimicrobial activities of EOs from Oreganum vulgare (oregano), Thymus vulgaris (thyme), Lippia graveolens (Mexican oregano), Cinnamomum zeylanicum (cinnamon), Zingiber officinale (ginger), Salvia officinalis (sage), Rosmarinus officinalis (rosemary) and Ocimum basilicum (basil) against Escherichia coli strains isolated from poultry (n=43) and cattle faeces (n=36). The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were determined for each isolate by using the broth microdilution technique, from the maximum concentration of 6400µg mL-1 of each OE tested. Antimicrobial activity was observed on the essential oils of oregano, mexican oregano, thymus and cinnamon. For all strains tested, regardless of their origin, the OES more and less effective as antimicrobial activity were oregano and cinnamon, respectively. These results confirm the antimicrobial potential of some EOs, which deserve further research, addressing the addition of essential oils in poultry and cattle feeding.

Avaliou-se a atividade antimicrobiana dos óleos essenciais (OEs) de Origanum vulgare (orégano), Thymus vulgaris (tomilho), Lippia graveolens (lípia), Zingiber officinale (gengibre), Salvia officinalis (sálvia), Rosmarinus officinalis (alecrim) e Ocimum basilicum (manjericão), bem como de frações majoritárias carvacrol, timol, cinamaldeído e cineol frente a 33 isolados de Staphylococcus spp oriundos de rebanhos leiteiros caprinos. A concentração inibitória mínima (CIM) e a concentração bactericida mínima (CBM) foram determinadas por meio da técnica de microdiluição em caldo. Observou-se atividade antimicrobiana para os OEs de orégano, lípia e tomilho, bem como para as frações majoritárias de carvacrol, timol e cinamaldeído. A ordem decrescente de atividade foi orégano = tomilho > lípia. As frações majoritárias carvacrol, timol e cinamaldeído evidenciaram melhor atividade do que os óleos essenciais e, dentre elas, carvacrol e cinamaldeído foram mais ativas que o timol.The antimicrobial activity of some essencial oils was evaluated as follows: Origanum vulgare (oregano), Thymus vulgaris (thyme), Lippia graveolens (Mexican oregano), Zingiber officinale (ginger), Salvia officinalis (sage), Rosmarinus officinalis (rosemary) and Ocimum basilicum (basil), as well as the majority constituents carvacrol, thymol, cinnamaldehyde and cineole against 33 Staphylococcus spp isolates from herds of dairy goats. The minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC) were determined for each isolate by using broth microdilution method. Antimicrobial activity observed on the essencial oils of oregano, mexican oregano, thymus, well as to majoritary constituents of carvacrol, thymol and cinnamaldehyde. The descending order of antimicrobial activity were oregano = thyme > mexican oregano. The majority constituents carvacrol, thymol, cinnamaldehyde presented themselves more active than the verified by the essencial oils. The majority constituents, carvacrol and cinnomaldehyde were equally more active than thymol.