Clinical and microbiological characteristics of dogs in sepsis in an academic veterinary hospital in the north of Paraná / Características clínicas e microbiológicas de cães em sepse de um hospital veterinário escola do norte do Paraná

Sepsis is a life-threatening organ dysfunction caused by a patient's unregulated response to an infectious process. In veterinary medicine, the exact incidence of sepsis is unknown. Early recognition of sepsis in critically ill patients is essential for rapid and effective therapeutic intervention. The present study aimed to apply the criteria of an adapted sepsis assessment protocol based on the Second International Consensus Definition for Sepsis and Septic Shock or Sepsis-2 of human medicine, in canine patients with suspected systemic inflammatory response syndrome (SIRS) and/or organ dysfunction, and to identify infectious agents as well as their antimicrobial resistance profile in the focus of infection, in the bloodstream and colonizing the rectal mucosa. Patients were evaluated for survival and severity of sepsis. Of the 37/42 dogs that met the sepsis criteria, six presented septic shock, 26 (70.2%) had at least two signs of SIRS, and sepsis with organ dysfunction was diagnosed in 27 (73%) dogs. The primary dysfunctions observed were decreased level of consciousness in 21/37 (56.8%), hyperlactatemia in 19/37 (51.4%), and hypoalbuminemia in 18/37 (48.6%). Two or more SIRS signs associated with hypotension and hypoalbuminemia were related to more than half of the deaths. The most frequent infectious focus was skin and soft tissue in 20/37 (54%), followed by organs and cavities in 8/37 (21.6%). The survival rate was 56.7%. Blood culture confirmed bacteremia in nine patients (24.3%), with a predominance of Gram-positive microorganisms (Staphylococcus intermedius, Streptococcus spp.) in 66.6% of dogs and one yeast (Candida glabrata). The most frequent bacteria in the focus of infection were gram-negative bacteria (46.2%), mainly Escherichia coli, Klebsiella pneumoniae, and Pseudomonas aeruginosa, in 19.5%, 14.6%, and 12.1%, respectively. We observed colonization by gram-negative bacteria such as E. coli-ESBL (31.5%), K. pneumoniae-ESBL (15.7%), and P. aeruginosa (15.7%), and the presence of ESBL bacteria was more associated with death when compared with other microorganisms. Vancomycin-resistant Enterococcus (VRE) were isolated from rectal mucosa in four dogs. Gram-negative microorganisms were the most frequent in both infections and colonization, and most of them were resistant to fluoroquinolones, sulfonamides, tetracyclines, and cephalosporins. Based on this information, it can be concluded that mortality due to sepsis in dogs was high. Due to the presence of multi-resistant bacteria, the use of antimicrobials should be judicious, suggesting the implementation of the same precautions used in human hospitals to prevent the spread of multi-resistant microorganisms.(AU)

A sepse é uma disfunção orgânica ameaçadora à vida, causada por uma resposta desregulada do hospedeiro à infecção e na medicina veterinária sua incidência exata é desconhecida. O reconhecimento precoce da sepse nos pacientes críticos é essencial para que a intervenção terapêutica seja rápida e eficaz. Assim, os objetivos do presente estudo foram aplicar os critérios de um protocolo de avaliação da sepse adaptado com base no Segundo Consenso Internacional para Sepse e Choque Séptico, ou Sepse-2, da medicina humana, em pacientes caninos com suspeita de infecção e/ou Síndrome da Resposta Inflamatória Sistêmica e/ou disfunção orgânica e identificar os agentes infecciosos bem como seu perfil de resistência a antimicrobianos no foco de infecção, na corrente sanguínea e colonizando a mucosa retal. Os pacientes foram avaliados quanto à sobrevivência e severidade da sepse. Dos 37/42 cães que se enquadraram nos critérios de sepse, seis estavam em choque séptico, 26 (70,2%) apresentaram pelo menos dois sinais de SIRS, e a sepse com disfunção orgânica foi diagnosticada em 27 (73%) cães. As principais disfunções verificadas foram diminuição do nível de consciência em 21/37 (56,8%), hiperlactatemia em 19/37 (51,4%) e hipoalbuminemia em 18/37 (48,6%). A presença de dois ou mais sinais de SIRS associados com hipotensão e hipoalbuminemia estiveram relacionadas com mais da metade dos óbitos. O foco infeccioso mais frequente foi pele e partes moles em 20/37 (54%) seguido por órgãos e cavidades em 8/37 (21,6%). A taxa de sobrevivência foi de 56,7%. Na hemocultura confirmou-se bacteremia em nove pacientes (24,3%), com predominância de microrganismos gram-positivos (Staphylococcus intermedius, Streptococcus spp.) em 66,6% dos cães e uma levedura (Candida glabrata). As bactérias mais frequentes no foco de infecção foram as gram-negativas (46,2%) principalmente Escherichia coli, Klebsiella pneumoniae e Pseudomonas aeruginosa, em 19,5%, 14,6% e 12,1% respectivamente. Foi constatada colonização por bactérias gram-negativas como E. coli-ESBL (31,5%), K. pneumoniae-ESBL (15,7%) e P. aeruginosa (15,7%), sendo que a colonização de cães por bactérias ESBL foi associada ao óbito quando comparada com outros microrganismos. Foram também isolados da mucosa retal Enterococcus resistentes à vancomicina (VRE) em quatro cães. Os microrganismos gram-negativos foram os mais frequentes, tanto nas infecções quanto nas colonizações e a maioria apresentava resistência à fluorquinolonas, sulfonamidas, tetraciclinas e cefalosporinas. Com base nestas informações, conclui-se que a mortalidade em decorrência da sepse em cães foi alta, e devido à presença de bactérias multirresistentes, o uso de antimicrobianos deve ser criterioso, sugerindo-se ainda a implantação das mesmas precauções utilizadas em hospitais humanos para evitar disseminação de microrganismos multirresistentes.(AU)

Deep tissue culture and hemoculture in dogs with wounds and sepsis / Cultura de tecido profundo e hemocultura em cães com feridas e sepse

Contaminated and infected wounds occur very frequently in veterinary medicine and can cause systemic inflammatory response syndrome, sepsis, and death. This study aimed to test the feasibility of collecting wound material by deep-tissue or punch biopsy for microbial culture, determine the frequency of bacteria in the wound(s) and blood cultures and the susceptibility of these microbes to antimicrobials, and evaluate clinical parameters that could be related to prognosis. Thirty dogs with wounds and signs of SIRS/sepsis were included in this study. Bacteria were isolated from all wounds and 41 bacterial isolates could be identified based on culture of the materials collected by punch biopsy; 53.66% of the isolates were gram-negative, mainly involving Pseudomonas aeruginosa, Klebsiella pneumoniae, and Enterococcus spp., and 46.34% were gram-positive bacteria such as Streptococcus spp., Enterococcus spp., and Staphylococcus spp. The survival rate was 66.67%. Based on blood culture analysis, we identified bacteremia in seven patients, predominantly of gram-negative bacteria, which negatively affected patient survival, as six dogs died. Hypoglycemia (≤60mg/dL) and severe hyperglycemia (≥180mg/dL) also negatively affected survival as 23.33% of the hypo/hyperglycemic dogs died. Factors such as blood lactate level at admission and hematocrit levels, and mean arterial pressure were not significantly correlated with death or survival of the dogs.(AU)

As feridas contaminadas e infectadas em cães ocorrem com grande frequência na medicina veterinária e podem causar síndrome da resposta inflamatória sistêmica, sepse e morte. Os objetivos do presente trabalho foram verificar a viabilidade da técnica de coleta de material da ferida por biópsia para realização de cultura microbiana, determinar a frequência das bactérias nas culturas das feridas e hemoculturas e a susceptibilidade destes agentes aos antimicrobianos, bem como avaliar parâmetros clínicos que pudessem ser relacionados ao prognóstico em 30 cães com feridas e sinais de SIRS/sepse. Foram isoladas bactérias de todas as feridas e a técnica de coleta de material para cultura por biópsia permitiu a obtenção de 41 agentes microbianos, sendo isoladas 53,66% bactérias Gram negativas e 46,34% Gram positivas, principalmente Pseudomonas aeruginosa, Klebsiella pneumoniae e Enterococcus spp. As bactérias gram positivas isoladas foram Streptococcus spp., Enterococcus spp. e Staphylococcus spp. A taxa de sobrevivência foi 66,67%. Na hemocultura constatou-se bacteremia em sete pacientes, com predominância de bactérias Gram negativas, o que influenciou negativamente na sobrevivência dos pacientes, pois seis cães vieram a óbito. A hipoglicemia (≤60mg/dL) ou hiperglicemia severa (≥180mg/dL), também influenciaram negativamente a sobrevivência, pois 23,33% dos pacientes hipo/hiperglicêmicos vieram a óbito. Já fatores como nível sérico de lactato na admissão do paciente, pressão arterial média (PAM) e hematócrito não apresentaram correlação estatística com o óbito ou sobrevivência destes pacientes.(AU)

Antibacterial Combination of Oleoresin from Copaifera multijuga Hayne and Biogenic Silver Nanoparticles Towards Streptococcus agalactiae.

BACKGROUND: Streptococcus agalactiae (group B Streptococcus - GBS) remains a leading cause of neonatal infections and an important cause of invasive infections in adults with underlying conditions. METHODS: This study evaluated for the first time the effect of an oleoresin collected from Copaifera multijuga Hayne (copaiba oil) alone or in combination with silver nanoparticles produced by green synthesis using Fusarium oxysporum (AgNPbio) against planktonic and sessile cells of GBS isolated from colonized women. RESULTS: Copaiba oil showed a dose-dependent bactericidal activity against planktonic GBS strains, including those resistant to erythromycin and/or clindamycin. Scanning and transmission electron microscopy of GBS treated with copaiba oil revealed morphological and ultrastructural alterations, displaying disruption of the cell wall and decreased electron density due to leakage of cytoplasmic content. Copaiba oil also exhibited antibacterial activity against biofilms of GBS strains, inhibiting their formation as well as the viability of mature biofilms. In addition, the combination of copaiba oil with AgNPbio resulted in a synergistic effect against planktonic cells and biofilm formation, reducing the minimal inhibitory concentration values of both compounds. No hemolytic activity was detected for both compounds. CONCLUSION: These results indicate the potential of copaiba oil, alone or in combination with AgNPbio, for the development of new alternative strategies for controlling GBS infections.

Antibacterial activity of extracellular compounds produced by a Pseudomonas strain against methicillin-resistant Staphylococcus aureus (MRSA) strains.

BACKGROUND: The emergence of multidrug-resistant bacteria is a world health problem. Staphylococcus aureus, including methicillin-resistant S. aureus (MRSA) strains, is one of the most important human pathogens associated with hospital and community-acquired infections. The aim of this work was to evaluate the antibacterial activity of a Pseudomonas aeruginosa-derived compound against MRSA strains. METHODS: Thirty clinical MRSA strains were isolated, and three standard MRSA strains were evaluated. The extracellular compounds were purified by vacuum liquid chromatography. Evaluation of antibacterial activity was performed by agar diffusion technique, determination of the minimal inhibitory concentration, curve of growth and viability and scanning electron microscopy. Interaction of an extracellular compound with silver nanoparticle was studied to evaluate antibacterial effect. RESULTS: The F3 (ethyl acetate) and F3d (dichloromethane- ethyl acetate) fractions demonstrated antibacterial activity against the MRSA strains. Phenazine-1-carboxamide was identified and purified from the F3d fraction and demonstrated slight antibacterial activity against MRSA, and synergic effect when combined with silver nanoparticles produced by Fusarium oxysporum. Organohalogen compound was purified from this fraction showing high antibacterial effect. Using scanning electron microscopy, we show that the F3d fraction caused morphological changes to the cell wall of the MRSA strains. CONCLUSIONS: These results suggest that P. aeruginosa-produced compounds such as phenazines have inhibitory effects against MRSA and may be a good alternative treatment to control infections caused by MRSA.