Effect of temperature on the lipolytic and proteolytic activity of Bacillus cereus isolated from dairy products

Bacillus cereus is a bacterium with deteriorating potential for dairy products, by being a psychrotrophic organism producer of lipases and proteases. This study evaluated the psychrotrophic behavior, lipolytic and proteolytic activity at 30°C, 10°C and 7°C of 86 strains of B. cereus lato sensu isolated from dairy products, marketed in Southern Brazil. It was also evaluated the optimal temperature for protease production. No strain grew at 7°C; but at 10°C, 84.9% of strains have grown. Only one strain had lipolytic activity at 30°C, and none at 7°C. At 10°C, 16.3% of strains produced lipases. All the strains presented proteolytic activity at 30°C; and at 10°C, 72.1% had this activity, and at 7°C, only 4.6%, an amount significantly lower (p < 0.05). The temperature of 20°C promoted the highest proteolytic activity, and at 10°C, the lowest activity. B. cereus can produce lipases and proteases at room and marginal chilling temperatures, causing technological defects in dairy products stored under these conditions. © 2008 IFRJ.

Molecular identification and antimicrobial susceptibility of Nocardia spp. isolated from bovine mastitis in Brazil

Nocardia spp. infections can cause severe damage to the mammary gland due to suppurative pyogranulomatous lesions and lack of clinical cure in response to conventional antimicrobial therapy. Although Nocardia infections are considered relatively uncommon in cows, there has been an apparent worldwide increase in the incidence of bovine mastitis caused by Nocardia spp, perhaps due to environmental transmission of this ubiquitous pathogen. The objectives of present study were to determine: (i) species distribution of 80 Nocardia isolates involved in bovine mastitis (based on molecular methods); and (ii) antimicrobial susceptibility pattern of all isolates from three geographical areas in Brazil. In this study, Nocardia nova (80%) was the most frequently isolated species, followed by Nocardia farcinica (9%). Additionally, Nocardia puris, Nocardia cyriacigeorgica, Nocardia veterana, Nocardia africana, and Nocardia arthritidis were detected using 16S rRNA sequencing. This is apparently the first report of N. puris, N. veterana, N. cyriacigeorgica, N. arthritidis and N. africana in association with bovine mastitis. Based on the disk diffusion test, isolates were most frequently resistant to cloxacillin (75%), ampicillin (55%) and cefoperazone (47%), whereas few Nocardia spp. were resistant to amikacin, cefuroxime or gentamicin. © 2013 Elsevier B.V. All rights reserved.

Molecular epidemiology and extended-spectrum β-lactamases production of klebsiella pneumoniae isolated from three dairy herds

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Effect of divalent metal ions on the activity and stability of β-galactosidase isolated from Kluyveromyces lactis

In this study, it was demonstrated that β-galactosidase can be deactivated and reactivated with EDTA and divalent metal ions. The enzyme was deactivated after 20 minutes in EDTA solution. Maximal deactivation at the lowest EDTA concentration (10-3 mol.L-1) occurred in the presence of Tris-HCl buffer (pH 7.0). The enzyme recovered 50% of its initial activity after 10 minutes at Mg2+concentrations higher than 0.1 mmol.L-1. Experimental concentrations of 0.1 mmol.L-1 Mn2+ and 1.0 mmol.L-1 Co2+ were sufficient to reactivate the enzyme to around 300% of the control activity for the Mn2+ ion and nearly 100% for the Co2+ ion. The enzyme gradually lost its activity when the Co2+ concentration was 10-2 mol.L-1. Ni2+ and Zn2+ were unable to restore the catalytic activity. Km app and Vmax app were 1.95 ± 0.05 mmol.L-1 and 5.40 ± 0.86x10-2 mmol.min-1.mg-1, with o-NPG as substrate. Optimal temperature and pH were 34oC and 7.5. The half-life (t1/2) at 30°C was 17.5 min for the holoenzyme and 11.0 min for the apoenzyme. With respect to pH variation, the apoenzyme proved to be more sensitive than the holoenzyme. Keywords: β-galactosidase. Divalent metallic ions. Enzyme activity. Stability. RESUMO Efeito de íons metálicos divalentes na atividade e estabilidade da β-galactosidase isolada de Kluyveromyces lactis Este estudo demonstra como a β-galactosidase pode ser desativada e reativada usando EDTA e íons metálicos divalentes. A enzima foi desativada após 20 minutos na presença de EDTA. Desativação máxima para a menor concentração de EDTA (10-3 mol.L-1) ocorreu na presença do tampão Tris-HCl. A enzima recuperou 50% de sua atividade inicial após 10 minutos na presença de Mg2+ em concentrações superiores a 0,1mmol.L-1. Concentrações de 10-4 e 10-3mol.L-1 de Mn2+ e Co2+ foram suficientes para reativar a enzima em 300% comparado ao controle de íons Mn2+ e aproximadamente 100% para íons Co2+. A enzima perdeu gradualmente a sua atividade quando a concentração foi de 10-2 mol.L-1. Ni2+ e Zn2+ foram incapazes de restabelecer a atividade catalítica. Km app e Vmax app foram 1,95 ± 0,05 mmol.L-1 e 5,40 ± 0,86 x 10-2 mmol.min-1.mg-1. A temperatura e pH ótimos foram 34ºC e 7,5. A meia vida da holoenzima foi de 17,5 min a 30ºC e para a apoenzima foi de 11,0 min a 30ºC. Quanto à variação de pH, a apoenzima provou ser mais sensível que a holoenzima. Palavras-chave: β-galactosidase. Íons metálicos divalentes. Atividade enzimática. Estabilidade.

NaOCl effect on biofilm produced by Staphylococcus aureus isolated from the milking environment and mastitis infected cows

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Beta-lactamase detection in Staphylococcus aureus and coagulase-negative Staphylococcus isolated from bovine mastitis

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Prevalence and phenotypic characterization of Enterococcus spp. isolated from food in Brazil

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Experimental infection of one-day-old chicks with Salmonella Serotypes Previously isolated from poultry facilities, wild birds, and swine

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Characterization, subsite mapping and N-terminal sequence of miliin, a serine-protease isolated from the latex of Euphorbia milii

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Effects of P-MAPA immunomodulator on Toll-like receptor 2, ROS, nitric oxide, MAPKp38 and IKK in PBMC and macrophages from dogs with visceral leishmaniasis

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Electrocardiographic evaluation of the degree of sedation and the isolated use of methadone in healthy dogs

Objective: The present study aimed to investigate the influence of methadone on cardiorespiratory parameters, electrocardiogram and clinical sedation in dogs. Further possible side effects are reported.Study designProspective experimental cross-over study.DogsEight, 1-4-year-old, various breeds of dogs of both genders weighing 9-36kg.MethodsEach dog was treated three times: methadone 0.3mgkg(-1) (M0.3), 0.5mgkg(-1) (M0.5) and 1.0mgkg(-1) (M1.0) intramuscularly. Respiratory rate, heart rate and arterial blood pressure were recorded as well as electrocardiographic evaluation of lead II. Clinical sedation in each treatment received a score (0-3) after drug administration and at 30minute intervals until scores and measurements returned to baseline values.ResultsA significant decrease in heart rate was seen with each dose of methadone and bradycardia (HR<60bpm) was noted in a few dogs at each dose. A clinically significant arrhythmia occurred in one dog at 1mgkg(-1) that required reversal with butorphanol. There was no significant difference in SAP, MAP and DAP between treatments. Some side effects such as salivation, defecation, vocalization and panting, after administration of methadone were observed. There were no differences in mean values of heart rate, P-wave and QRS complex duration and QT interval between treatments.Conclusion and clinical relevanceMethadone administration was associated with panting and a decrease in heart rate at all doses tested in this study. The cardiac rhythm should be monitored carefully in dogs when methadone is administered on its own, especially at higher doses.

Enzymatic activity profile of a Brazilian culture collection of Candida albicans isolated from diabetics and non-diabetics with oral candidiasis

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Diagnostic accuracy of semi-quantitative and quantitative culture techniques for the diagnosis of catheter-related infections in newborns and molecular typing of isolated microorganisms

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Diversity of Staphylococcus species and prevalence of enterotoxin genes isolated from milk of healthy cows and cows with subclinical mastitis

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)