Activity of biogenic silver nanoparticles against isolates of Prototheca species from bovine mastitis.

Prototheca spp. cause numerous infections in a wide variety of species, including treatment-unresponsive mastitis. Thus, the search for an effective therapy is essential. Silver nanoparticles are compounds with high therapeutic potential. This study aimed to evaluate the susceptibility profile and morphological changes in Prototheca spp. treated with biogenic silver nanoparticles (Bio-AgNP). The algaecide activity was evaluated in microplates by microdilution method, resulting in a MIC50 of 30 µg ml-1 and a MIC90 of 60 µg ml-1 . Scanning electron microscopy demonstrated changes in the surface of Prototheca bovis cells following treatment. The algaecide activity of Bio-AgNP suggests a therapeutic potential as a novel approach for the control of Prototheca spp. in bovine mastitis.

In vitro algicidal effect of guanidine on Prototheca zopfii genotype 2 strains isolated from clinical and subclinical bovine mastitis.

Prototheca species have increasingly been reported to be opportunistic pathogens that cause mastitis in dairy herds, and it poses an emergent problem because at present, there are no effective therapies for the treatment of protothecal mastitis. This study investigated the in vitro algicidal effect of guanidine on 75 Prototheca zopfii genotype 2 strains isolated from 75 cases of clinical and subclinical bovine mastitis. All strains were susceptible to guanidine in vitro with minimal algaecide concentrations ranging from 0·001 to 0·035%. Guanidine is known to have a high microbicidal effect and is considered to be a new generation microbicidal compound. It is not toxic to human mucous membranes and conjunctivas at low concentrations and has been used as a disinfectant in swimming pools and as an antiseptic for human wounds. The algicidal action of guanidine at low concentrations indicates that it could be an alternative disinfectant or antiseptic for cleaning of the dairy environment and milking equipment, in pre- and postdipping solutions, in the chemical dry therapy of bovine teats and even in the intramammary therapy of P. zopfii infections. This is the first report of the in vitro algicidal effect of guanidine on P. zopfii strains of animal origin. SIGNIFICANCE AND IMPACT OF THE STUDY: Prototheca zopfii genotype 2 is an opportunistic pathogen of bovine mastitis. To date, no effective therapies against protothecal mastitis have been developed. The in vitro algicidal effect of guanidine on 75 P. zopfii genotype 2 strains isolated from cows revealed that all of the isolates were susceptible to the compound at low concentrations, which indicates that guanidine may be used as an antiseptic/disinfectant for dairy milking equipment, in pre- and postdipping solutions, and as a chemical dry therapy or an intramammary therapy. This study describes the in vitro algicidal effect of guanidine on P. zopfii for the first time.

Fluorescence spectroscopy in renal ischemia and reperfusion: noninvasive evaluation of organ viability.

BACKGROUND: Damage provoked by ischemia in renal transplants is difficult to quantify. To determine whether a donated organ is fit for transplantation. We sought to correlate the findings of fluorescence spectroscopy (FS) with histologic evidence of ischemic injury and organ viability. METHODS: Kidneys of 33 rats were submitted to FS of the upper and lower poles as well as the middle third. Excitation was generated by the laser's wavelengths of 408, 442, and 532 nm. Rats were randomized into groups with the 30, 60, and 120 minutes warm ischemia before analysis by FS, that was repeated at 5 minutes after reperfusion. RESULTS: FS results in the reperfusion phase correlated with ischemia time and degree of histologic injury. After 60 or 120 minutes of ischemia, the excitation lasers of 532 and 442 nm resented a significant negative correlation coefficient with the histological grade (r = -0.61 and r = -0.73, respectively). CONCLUSIONS: There was a strong correlation between FS and histologic changes only in the reperfusion phase after renal ischemia. The method was thus unable to assess the viability of organs before transplantation.

Evaluation by fluorescence spectroscopy of the most appropriate renal region for obtaining biopsies: a study in the rat.

INTRODUCTION: Renal puncture biopsies are directed at the lower poles of the organ to decrease the risk of hemorrhage and complications. OBJECTIVES: To evaluate by fluorescence spectroscopy (FS) the most appropriate renal region (in terms of metabolic changes) to obtain a biopsy. MATERIALS AND METHODS: The kidneys of 33 Rattus norvegicus rats were submitted to FS detection in the upper and lower poles and in the middle third. Excitations were generated with lasers at wavelengths of 408, 442, and 532 nm. Animals were divided at random into groups of warm ischemia (30, 60, and 120 minutes), whose kidneys were again analyzed by FS, as well as after 5 minutes of reperfusion using the same excitation beams in the same renal regions. Then the kidneys underwent histologic preparation and examination. RESULTS: The middle third area of the rat's kidneys proved to be significantly more sensitive to ischemic and reperfusion changes than the renal poles, as determined by FS (P < .001). CONCLUSIONS: The middle third of the kidney was the most appropriate site for a renal biopsy to monitor a transplanted organ.

Prion disease (PrP-A117V) presenting with ataxia instead of dementia.

Gerstmann-Sträussler-Scheinker disease (GSS) is caused by several different point mutations of the prion protein (PrP) gene, each of which generally produces a distinct clinical phenotype. An ataxic form of GSS is genetically linked to a mutation at codon 102 (CCG-->CTG) leading to the substitution of leucine for proline, while a "telencephalic" variant of GSS, in which dementia is the predominant symptom and ataxia is minimal, has been described in two kindreds with a mutation at codon 117 (GCA-->GTG) resulting in the substitution of valine for alanine. In this report, we present a family with ataxic GSS that has, however, the same mutation at codon 117 as is present in the telencephalic variant of GSS. Other than an additional silent mutation (GCA-->GCG) at codon 117 on the normal allele, there were no other mutations detected. At the polymorphic codon 129, valine was encoded by both alleles in the proband that we studied. Why this family with prion disease (PrP-A117V) should present with ataxia instead of dementia, which was found in two previously identified families with the same PrP gene mutation, remains to be established.

Metabolism of glucose, glutamine and pyruvate in lymphocytes from Walker 256 tumor-bearing rats.

This study examined the effect of Walker 256 tumor growth in vivo on the metabolism of glucose, glutamine and pyruvate in lymphocytes. A comparison between the metabolism of Walker 256 tumor cells obtained in vivo with that of lymphocytes was also carried out. Lymphocytes and tumor cells were isolated and incubated for 1 h for the following measurements: lactate production from glucose (5.6 mM) and pyruvate (3 mM), glutamate and aspartate formation from glutamine (3 mM) and decarboxylation of [U-14C]-glucose, [U-14C]-glutamine, [1-14C]-pyruvate and [3-14C]-pyruvate. The presence of the tumor increased lactate production (2.7-fold from glucose and 2-fold from pyruvate), decarboxylation of [U-14C]-glucose (3.7-fold) and [1-14C]-pyruvate (4.4-fold) and the formation of aspartate (6.3-fold) and glutamate (4.6-fold) from glutamine. The conversion of glucose to lactate and CO2 was higher in tumor cells as compared to lymphocytes. Tumor cells also showed a higher production of glutamate and an 8-fold increased decarboxylation rate of [U-14C]-glutamine in tumor cells, which was more active than that of lymphocytes even from tumor-bearing rats. Tumor growth stimulated glucose and glutamine metabolism in lymphocytes; however, the importance of this fact for the function of these cells remains to be elucidated.