The Th2 Response and Alternative Activation of Macrophages Triggered by Strongyloides venezuelensis Is Linked to Increased Morbidity and Mortality Due to Cryptococcosis in Mice.

Cryptococcosis is a systemic mycosis that causes pneumonia and meningoencephalitis. Strongyloidiasis is a chronic gastrointestinal infection caused by parasites of the genus Strongyloides. Cryptococcosis and strongyloidiasis affect the lungs and are more prevalent in the same world regions, i.e., Africa and tropical countries such as Brazil. It is undeniable that those coincidences may lead to the occurrence of coinfections. However, there are no studies focused on the interaction between Cryptococcus spp. and Strongyloides spp. In this work, we aimed to investigate the interaction between Strongyloides venezuelensis (Sv) and Cryptococcus gattii (Cg) in a murine coinfection model. Murine macrophage exposure to Sv antigens reduced their ability to engulf Cg and produce reactive oxygen species, increasing the ability of fungal growth intracellularly. We then infected mice with both pathogens. Sv infection skewed the host's response to fungal infection, increasing lethality in a murine coinfection model. In addition to increased NO levels and arginase activity, coinfected mice presented a classic Th2 anti-Sv response: eosinophilia, higher levels of alternate activated macrophages (M2), increased concentrations of CCL24 and IL-4, and lower levels of IL-1ß. This milieu favored fungal growth in the lungs with prominent translocation to the brain, increasing the host's tissue damage. In conclusion, our data shows that primary Sv infection promotes Th2 bias of the pulmonary response to Cg-infection and worsens its pathological outcomes.

Randomized, phase 1/2, double-blind pioglitazone repositioning trial combined with antifungals for the treatment of cryptococcal meningitis - PIO study.

BACKGROUND: Cryptococcosis affects more than 220,000 patients/year, with high mortality even when the standard treatment [amphotericin B (AMB), 5-flucytosin (5-FC) and fluconazole] is used. AMB presents high toxicity and 5-FC is not currently available in Brazil. In a pre-clinical study, pioglitazone (PIO - an antidiabetic drug) decreased AMB toxicity and lead to an increased mice survival, reduced morbidity and fungal burden in brain and lungs. The aim of this trial is to evaluate the efficacy and safety of PIO combined with standard antifungal treatment for human cryptococcosis. METHODS: A phase 1/2, randomized, double blind, placebo-controlled trial will be performed with patients from Belo Horizonte, Brazil. They will be divided into three groups (placebo, PIO 15 mg/day or PIO 45 mg/day) and will receive an additional pill during the induction phase of cryptococcosis' treatment. Our hypothesis is that treated patients will have increased survival, so the primary outcome will be the mortality rate. Patients will be monitored for survival, side effects, fungal burden and inflammatory mediators in blood and cerebrospinal fluid. The follow up will occur for up 60 days. CONCLUSIONS: We expect that PIO will be an adequate adjuvant to the standard cryptococcosis' treatment. TRIAL REGISTRATION: ICTRP/WHO (and International Clinical Trial Registry Plataform (ICTRP/WHO) (http://apps.who.int/trialsearch/Trial2.aspx?TrialID=RBR-9fv3f4), RBR-9fv3f4 (http://www.ensaiosclinicos.gov.br/rg/RBR-9fv3f4). UTN Number: U1111-1226-1535. Ethical approvement number: CAAE 17377019.0.0000.5149.

Efficacy of novel antiseptic product containing essential oil of Lippia origanoides to reduce intramammary infections in cows.

BACKGROUND AND AIM: The use of antimicrobials in the control of mastitis is of concern in public health due to their inefficiency in targeting microorganisms. Studies with medicinal plants have risen as an alternative to the use of conventional products. The objective of this study was to evaluate the efficacy of an experimental disinfectant based on the essential oil (EO) from Lippia origanoides in preventing the development of new intramammary infections (IMI) in Holstein cows. MATERIALS AND METHODS: The conventional protocol of pre- and post-milking was used and the control (Conventional treatment [CNV]) and experimental (Experimental treatment [PEX]) products containing EO at 120 µL/mL were applied by immersion. Individual milk samples were analyzed using sheep blood agar methodologies and biochemical tests. The efficiency of the treatment was defined by the presence or absence of Staphylococcus aureus, coagulase-negative Staphylococcus, and Streptococcus spp. RESULTS: There were no clinical and subclinical mastitis cases, no lesions in the mucosal of teats, nor dirt score between groups in this study. Both treatments did not influence the occurrence of IMI. CONCLUSION: The results revealed that PEX acts efficiently against microorganisms compared to the disinfection by the conventional product demonstrating the efficacy of the alternative product on the prevention of new IMIs in dairy cows.

Characterization and molecular epidemiology of Staphylococcus aureus strains resistant to beta-lactams isolated from the milk of cows diagnosed with subclinical mastitis.

BACKGROUND AND AIM: The term ESKAPE, recognized by the WHO, is an acronym, which refers to the pathogens Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter spp., which is extremely virulent and multidrug-resistant. Although the term is used to designate nosocomial pathogens, in a milking environment, strains of Methicillin-resistant S. aureus have been isolated from cattle diagnosed with clinical and subclinical mastitis. Resistant strains may be involved in the transfer of genes conferring resistance to beta-lactam antimicrobials among the species of microorganisms related to mastitis etiology. This study aimed to trace the phenotypic and genotypic profiles of susceptibility to beta-lactams in S. aureus isolated from milk of cattle diagnosed with subclinical mastitis obtained from different rural properties located in the North of Minas Gerais State, Brazil. MATERIALS AND METHODS: Sixteen microorganisms previously identified as S. aureus isolated from milk of cattle diagnosed with subclinical mastitis were submitted to matrix-assisted laser desorption/ionization-time-of-flight (MALDI-TOF), mass spectrometry, and polymerase chain reaction (PCR) analysis for microbial species confirmation. The S. aureus beta-lactams antimicrobial phenotypic resistance profile was investigated by disk diffusion method. PCR methods were also performed to investigate the S. aureus genotypic beta-lactams resistance profile. For this purpose, bla Z, mec A, mec ALGA251, bla Oxa23, and bla KPC genes were screened among S. aureus isolates. The genetic diversity of S. aureus by fingerprint random amplified polymorphic DNA (RAPD)-PCR was also performed in this study. RESULTS: All isolates showed phenotypic resistance to at least three beta-lactams, among which was meropenem. None of the isolates tested positive for the genes mec ALGA251, bla Oxa23, and bla KPC; however, the presence of the genes bla Z and mecA was detected among the isolates. The fingerprint analysis divided isolates into two distinct groups and 15 different subgroups. Despite the presence of clonality among the isolates, the PCR-RAPD analysis unveiled a heterogeneous profile with genetic diversity among the S. aureus isolates. CONCLUSION: In this study, we identified beta-lactams resistant S. aureus strains isolated from the milk of cows diagnosed with subclinical mastitis. The S. aureus beta-lactams resistance was investigated using a phenotypic and genotypic approach. We believe that molecular epidemiology, improved knowledge, and genetic basis of resistance to beta-lactams might assist in asserting guidelines for better management practices of dealing with subclinical mastitis and mapping of origin of resistant pathogens in the studied Brazilian area.

Atividade antisséptica do óleo essencial de Lippia origanoides Cham. (Alecrim-pimenta) na presença de leite bovino / Antiseptic activity of the essential oil of Lippia origanoides in the presence of bovine milk

Objetivou-se determinar atividade antisséptica do óleo essencial de Lippia origanoides na presença de leite bovino. A composição química do óleo essencial de alecrim pimenta foi determinada por cromatografia gasosa acoplada à espectrometria de massas (CG-EM), sendo detectados 53 compostos, dos quais 16 foram identificados (>0,1% área total). O carvacrol (32,7%), p-cimeno (23%), timilmetil éter, cariofileno (7,98%) e o γ-terpineno (5,40%) foram os componentes mais abundantes. A concentração inibitória mínima para Staphylococcus aureus ATCC 25923 e E. coli ATCC 8739 foi de 60µL/mL, enquanto para Salmonella Choleraesuis ATCC 35640 foi de 90µL/mL. A concentração bactericida mínima foi de 120µL/mL para as três bactérias. Avaliou-se a atividade antisséptica do óleo essencial na concentração de 120µL/mL na presença de leite bovino sobre as mesmas cepas bacterianas. O óleo em estudo apresentou efeito inibitório do crescimento das cepas em diferentes tempos de ação (p<0,05). S. aureus apresentou maiores índices de inibição após 5 min de contato e Escherichia coli e Salmonella Choleraesuis após 15 min. O óleo essencial de alecrim-pimenta revelou ser um potencial antimicrobiano natural, mesmo na presença de matéria orgânica constituída de uma matriz nutricional complexa que é o leite bovino.(AU)

This study aimed to determine the antiseptic activity of the essential oil of Lippia origanoides in the presence of bovine milk. The essential oil chemical composition was determined by gas chromatography-mass spectrometry (CG EM) 53 compounds were detected, of which 16 were identified (>0.1% total area). The carvacrol (32.7%), p-cymene (23%), thymil methyl ether (10.03%), caryophyllene (7.98 %) and γ-terpinene (5.40%) were the most abundant components. The minimum inhibitory concentration for Staphylococcus aureus ATCC 25923 and Escherichia coli ATCC 8739 was 60uL/mL, as for this Salmonella Choleraesuis ATCC 35640 was 90uL/mL. The minimum bactericidal concentration was 120µL/mL for all three bacteria. We evaluated the antiseptic activity of the essential oil in the concentration of 120µL/mL in the presence of bovine milk for the same bacterial strains. The oil under study It showed an inhibitory effect of growth of the strains in different action times (p <0.05). S. aureus showed higher inhibition rates after 5 min of contact and E. coli and Salmonella Choleraesuis after 15 min. The essential oil Lippia origanoides proved to be a natural antimicrobial potential even in the presence of organic matter consists of a complex nutritional matrix is bovine milk.(AU)