To verify the occurrence of caseous lymphadenitis in sheep and goats on farms of Pernambuco, Brazil, and in animals slaughtered in two Brazilian cities (Petrolina/PE and Juazeiro/BA), and to characterize the susceptibility profile of Corynebacterium pseudotuberculosis to disinfectants and antimicrobials, and its relationship with biofilm production were the objectives of this study. 398 samples were tested for sensitivity to antimicrobial drugs, disinfectants, and biofilm production. Among the 108 samples collected on the properties, 75% were positive for C. pseudotuberculosis. Slaughterhouse samples indicated an occurrence of caseous lymphadenitis in 15.66% and 6.31% for animals slaughtered in Petrolina and Juazeiro respectively. With respect to antimicrobials, the sensitivity obtained was 100% for florfenicol and tetracycline; 99.25% for enrofloxacin, ciprofloxacin and lincomycin; 98.99% for cephalothin; 98.74% for norfloxacin and sulfazotrim; 97.74% for gentamicin; 94.22% for ampicillin; 91.71% for amoxicillin; 91.21% for penicillin G; 89.19% for neomycin and 0% for novobiocin. In analyzes with disinfectants, the efficiency for chlorhexidine was 100%, 97.20% for quaternary ammonium, 87.40% for chlorine and 84.40% for iodine. 75% of the isolates were weak or non-biofilm producers. For the consolidated biofilm, found that iodine decreased biofilm formation in 13 isolates and quaternary ammonia in 11 isolates. The reduction of the biofilm formation was observed for iodine and quaternary ammonium in consolidated biofilm formation in 33% and 28% of the isolates, respectively. The results of this study highlight the importance of establishing measures to prevent and control the disease.

As antibiotics are ineffective when used to treat caseous lymphadenitis, the surgical excision of lesions is often required. Iodine solution (10%) is currently the choice for the post-surgical treatment; however, it may cause histotoxicity. Propolis are resinous substances composed by a mixture of different plants parts and molecules secreted by bees. As green propolis has already proven to possess anti-bacterial and wound healing properties, this study aimed to evaluate the use of a green propolis-based ointment as a therapeutic agent for the post-surgical treatment of caseous lymphadenitis. The caseous lesions of 28 sheep were surgically excised before dividing animals into two groups: (1) iodine-treated animals and (2) sheep treated with an ointment made with a previously characterized green propolis extract. Clinical data of animals, size of the scar area, the presence of moisture and secretion in the surgical wound, the humoral immune response against the bacterium and the susceptibility of C. pseudotuberculosis clinical isolates to the green propolis extract were analyzed. The green propolis-treated group presented complete healing of the surgical wound 1 week before the iodine-treated group. Additionally, animals treated with the green propolis ointment had fewer cases of wound secretion, but it was not statistically different from the iodine-treated group. No clinical signs indicating green propolis toxicity or other side effects were found, associated with a faster and more organized hair recovery by propolis use. The green propolis extract was able to inhibit the growth of 23 from the 27 C. pseudotuberculosis clinical isolates, with minimum inhibitory and minimum bactericide concentrations ranging from 01 to 08 mg/mL, and did not interfere with the humoral immune response against the bacterium. In addition, green propolis was able to inhibit biofilm formation by four of the C. pseudotuberculosis clinical isolates. We concluded that green propolis is a promising therapeutic agent to be used in the post-surgical treatment of caseous lymphadenitis in small ruminants due to its effects on surgical wound healing, hair recovery, inhibition of wound contamination and bacterial growth.

A busca por tratamentos fitoterápicos intensificou-se nas últimas décadas. O uso abusivo das drogas antimicrobianas, a seleção de bactérias resistentes e as condições de manejo inadequadas são temas de impacto na medicina veterinária. Tendo em vista o acima descrito, objetivou-se avaliar a atividade antibacteriana de extratos etanólicos de plantas do bioma caatinga, contra patógenos de interesse veterinário. Foram utilizados seis extratos etanólicos de Amburana cearensis A.C.Smith, a Selaginella convoluta Arn.(Spring), a Hymenaea courbaril L., a Neoglaziovia variegata (Arruda) Mez., Bromelia laciniosa Mart. e Encholirium spectabile Mart. A atividade antibacteriana destes extratos foi testada contra bactérias gramnegativas e positivas. Determinou-se a concentração bactericida mínima (CBM) para cada extrato. Os ensaios foram realizados em triplicata. Para os extratos avaliados foram encontradas atividades antibacterianas nos gêneros estudados com exceção de Proteus spp., Nocardia spp., Staphylococcus caprae e Streptococcus agalactiae. A umburana (A. cearensis) e o caroá (Neoglaziovia variegata) mostraram os menores valores de CBM. Considerando o baixo custo da fitoterapia e a atividade das plantas do bioma caatinga, outros estudos acerca da atividade in vivo e da caracterização fitoquímica tornam-se necessários.

BACKGROUNDLaser 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. OBJECTIVETo assess the effect of low intensity laser on the rate of bacterial contamination in infected wounds in the skin of rats. METHODSAn 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 <0.05. RESULTSThe statistical analysis of median values showed that the groups submitted to laser treatment had low bacterial proliferation. CONCLUSIONThe 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.

RESUMOAs bactérias do gênero Aeromonas spp. são considerados como patógenos oportunistas carreadores de múltiplos fatores de virulência. O fenômeno da resistência aos antimicrobianos e metais pesados constitui outro problema, podendo ocorrer por diferentes fatores, dentre eles o uso indiscriminado de agentes antimicrobianos, poluição ambiental e a presença de mecanismos de resistência, como bombas de efluxo, sendo que muitos destes podem ser transmitidos por elementos genéticos móveis como os plasmídeos. O objetivo desse artigo é fazer uma revisão bibliográfica sobre os fatores de virulência, resistência a antimicrobianos e metais pesados, bem como os mecanismos que podem intervir nessa resistência e sua transferência entre bactérias do gênero Aeromonas. Os estudos dos fatores envolvidos no mecanismo de surgimento da resistência, aliado aos estudos de biologia molecular, fornecem subsídios para elaboração de métodos de controle e profilaxia dessas enfermidades de impacto aos organismos aquáticos, seres humanos e meio ambiente. PALAVRAS-CHAVE: Micro-organismos, fatores de virulência, plasmídeo, bomba de efluxo.ABSTRACT AEROMONAS SPP.: VIRULENCE FACTORS AND RESISTANCE PATTERNS TO ANTIMICROBIAL AND HEAVY METALS. Bacteria from Aeromonas spp. genus are considered as opportunistic pathogens that can carry many virulence factors. The resistance phenomenon to antimicrobial drugs and heavy metals is another problem and may occurs by several factors, as the indiscriminate usage, environmental pollution and the presence of resistance determinants as efflux pumps, being many of them transmitted by genetic mobile elements as plasmid. The purpose of this article is to review about the virulence factors, antimicrobial and heavy metal resistance, as well as the mechanisms that can help its transfer among Aeromonas genus bacteria. Studies of the resistance mechanisms associated to molecular biology could produce information about control and prophylaxis methods to this disease of impact on aquatic organisms, man and environment.

ObjectivePrevious works defining antigens that might be used as vaccine targets against Corynebacterium pseudotuberculosis, which is the causative agent of sheep and goat caseous lymphadenitis, have focused on secreted proteins produced in a chemically defined culture media. Considering that such antigens might not reflect the repertoire of proteins expressed during infection conditions, this experiment aimed to investigate the membrane-associated proteins with pathogenic potential expressed by C. pseudotuberculosis grown directly in animal serum.ResultsIts membrane-associated proteins have been extracted using an organic solvent enrichment methodology, followed by LC–MS/MS and bioinformatics analysis for protein identification and classification. The results revealed 22 membrane-associated proteins characterized as potentially pathogenic. An interaction network analysis indicated that the four potentially pathogenic proteins ciuA, fagA, OppA4 and OppCD were biologically connected within two distinct network pathways, which were both associated with the ABC Transporters KEGG pathway. These results suggest that C. pseudotuberculosis pathogenesis might be associated with the transport and uptake of nutrients; other seven identified potentially pathogenic membrane proteins also suggest that pathogenesis might involve events of bacterial resistance and adhesion. The proteins herein reported potentially reflect part of the protein repertoire expressed during real infection conditions and might be tested as vaccine antigens.Electronic supplementary materialThe online version of this article (10.1186/s13104-018-3180-5) contains supplementary material, which is available to authorized users.

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.

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.