RESUMO
Objective.To present an innovative approach for the design of a 3D mammographic phantom for medical equipment quality assessment, estimation of the glandular tissue percentage in the patient's breast, and emulation of microcalcification (µC) breast lesions.Approach.Contrast-to noise ratio (CNR) measurements, as well as spatial resolution and intensity-to-glandularity calibrations under mammography conditions were performed to assess the effectiveness of the phantom. CNR measurements were applied to different groups of calcium hydroxyapatite (HA) and aluminum oxide (AO)µCs ranging from 200 to 600µm. Spatial resolution was characterized using an aluminum plate contained in the phantom and standard linear figures of merit, such as the line spread function and modulation transfer function (MTF). The intensity-to-glandularity calibration was developed using an x-ray attenuation matrix within the phantom to estimate the glandular tissue percentage in a breast with a compressed thickness of 4 cm.Main results.For the prototype studied, the minimum confidence level for detecting HAµCs is 95.4%, while for AOµCs is above 68.3%. It was also possible to determine that the MTF of the commercial mammography machine used for this study at the Nyquist frequency is 41%. Additionally, a one-to-one intensity-to-glandularity calibration was obtained and verified with Monte-Carlo simulation results.Significance.The phantom provides traditional arrangements presented in accreditation phantoms, which makes it competitive with available devices, but excelling in regarding affordability, modularity, and inlays distribution. Moreover, its design allows to be positioned in close proximity to the patient's breast during a medical screening for a simultaneous x-ray imaging, such that the features of the phantom can be used as reference values to specify characteristics of the real breast tissue, such as proportion of glandular/adipose composition and/orµC type and size lesions.
Assuntos
Mama , Mamografia , Humanos , Mamografia/métodos , Mama/diagnóstico por imagem , Simulação por Computador , Imagens de Fantasmas , Raios XRESUMO
The use of yeasts as a feed supplement for cattle can promote animal development and performance. However, for the positive results to be consistent, strains with probiotic properties must be selected. The objective of this study was to isolate and identify yeasts present in the bovine feces and evaluate their probiotic potential together with strains previously isolated from the rumen (preliminary study). A total of 193 isolates were studied, including 139 isolates (19 species) from fecal samples from 11 different animals (Bos taurus and Bos indicus) and 54 strains previously isolated from rumen fluid (Bos taurus). The yeast population in the feces ranged from 3.51 to 4.99 log CFU/g, with Candida pararugosa being the most abundant (isolated from the feces of six samples analysed). Isolates were selected that had negative results in the safety tests (hemolytic activity, DNAse, and gelatinase) and had percentages greater than 35 and 70% for hydrophobicity and auto-aggregation, respectively. In addition, selected isolates had percentages greater than 77.7 and 74.7% for coaggregation with pathogenic strains of Escherichia coli and Clostridium perfringens, respectively. The isolates with percentage growth at 39 °C greater than 64.6% and viability greater than 96.7% were selected for survival testing under bovine gastrointestinal conditions. After the tests, the seven best isolates were selected, belonging to the species Candida pararugosa (L60, CCMA 928 and CCMA 930) and Pichia kudriavzevii (L97, L100, CCMA904, CCMA 907). The selected isolates were exopolysaccharide producers. Based on the results of the evaluated properties, the seven selected isolates were classified as potential probiotics for cattle.
Assuntos
Probióticos , Saccharomyces cerevisiae , Bovinos , Animais , Trato Gastrointestinal , Fezes , Escherichia coliRESUMO
Cellulolytic micro-organisms are potent silage inoculants that decrease the fibrous content in silage and increase the fibre digestibility and nutritional value of silage. This study aimed to evaluate the effects of Bacillus subtilis CCMA 0087 and its enzyme ß-glucosidase on the nutritional value and aerobic stability of corn silage after 30 and 60 days of storage. We compared the results among silage without inoculant (SC) and silages inoculated with B. subtilis 8 log10 CFU per kg forage (SB8), 9 log10 CFU per kg forage (SB9) and 9·84 log10 CFU per kg forage + ß-glucosidase enzyme (SBE). No differences were observed in the levels of dry matter, crude protein and neutral detergent fibre due to the different treatments or storage times of the silos. Notably, the population of spore-forming bacteria increased in the SB9-treated silage. At 60 days of ensiling, the largest populations of lactic acid bacteria were found in silages treated with SB8 and SBE. Yeast populations were low for all silages, irrespective of the different treatments, and the presence of filamentous fungi was observed only in the SBE-treated silage. Among all silage treatments, SB9 treatment resulted in the highest aerobic stability.
Assuntos
Ração Animal/microbiologia , Bacillus subtilis/metabolismo , Silagem/microbiologia , Silagem/normas , Zea mays/microbiologia , Ração Animal/análise , Ração Animal/normas , Fibras na Dieta/metabolismo , Fungos/isolamento & purificação , Lactobacillus/metabolismo , Leveduras/isolamento & purificaçãoRESUMO
The main challenge of ensiling is conserving the feed through a fermentative process that results in high nutritional and microbiological quality while minimizing fermentative losses. This challenge is of growing interest to farmers, industry and research and involves the use of additives to improve the fermentation process and preserve the ensiled material. Most studies involved microbial additives; lactic acid bacteria (LAB) have been the focus of much research and have been widely used. Currently, LABs are used in modern and sustainable agriculture because of their considerable potential for enhancing human and animal health. Although the number of studies evaluating LABs in silages has increased, the potential use of these micro-organisms in association with silage has not been adequately studied. Fermentation processes using the same strain produce very different results depending on the unique characteristics of the substrate, so the choice of silage inoculant for different starting substrates is of extreme importance to maximize the nutritional quality of the final product. This review describes the current scenario of the bioprospecting and selection process for choosing the best LAB strain as an inoculant for ensiling. In addition, we analyse developments in the fermentation process and strategies and methods that will assist future studies on the selection of new strains of LAB as a starter culture or inoculant.
Assuntos
Lactobacillales/isolamento & purificação , Valor Nutritivo , Silagem/microbiologia , Silagem/normas , Animais , Bioprospecção , Fermentação , Lactobacillales/classificação , Lactobacillales/metabolismoRESUMO
Advances in micro-organism identification techniques have resulted in increased knowledge of the diversity of prokaryotes and eukaryotes in silage. Such knowledge has enhanced the understanding of how fermentation occurs in forage crops with different characteristics and how the process can be improved to enhance silage quality. Undesirable micro-organisms can grow in silage when fermentation does not occur properly. Such micro-organisms may be pathogenic and/or produce toxic metabolic compounds; however, information on the consequences of these metabolites on the health of animals that consume silage is still lacking. The major challenge of ensilage is to produce high-quality feed that is nutritional, sanitary and stable, with a high dry matter recovery rate, in a process involving no interventions during fermentation and considerable variation in the characteristics of the substrates. It is important to note that each substrate has particularities and that we can only improve fermentation if we fully understand microbial diversity. This review is intended to update information related to the fermentation profile of silage, focusing on microbial diversity.
Assuntos
Microbiota , Silagem/microbiologia , Animais , Fermentação , Microbiota/genética , Silagem/análise , Silagem/normasRESUMO
AIMS: The aim was to isolate, identify and characterize yeasts present in rumen fluid and to select strains showing potential as probiotics. METHODS AND RESULTS: Rumen fluid was sampled from 4 herds of dairy and beef cattle and 77 yeast isolates were identified. Initial screening was based on the capacity to maintain viability in a medium with different ruminal conditions. A second screening in fresh rumen fluid to assess the growth of inoculated yeasts and evaluate in vitro neutral detergent fibre digestibility (NDF-D), pH and acid accumulation was conducted. The yeast population ranged from 3·84 to 6·76 log10 CFU per ml. The main species of yeast found were Pichia kudriavzevii, Candida rugosa, C. pararugosa, C. ethanolica and Magnusiomyces capitatus. Strains CCMA 933 (C. rugosa) and CCMA 970 (C. pararugosa) showed greater ability to survive in ruminal fluid and stimulated the production of acids. Isolate CCMA 967 (C. ethanolica) survived and improved the NDF-D. CONCLUSION: Pichia kudriavzevii was the dominant yeast found in the cattle herds. Strains CCMA 933, CCMA 970 and CCMA 967 showed properties that could be useful as potential probiotics for cattle. SIGNIFICANCE AND IMPACT OF THE STUDY: This study was the first to select yeasts from the rumen fluid, with the potential to be used as probiotic, based on the ruminal conditions.
Assuntos
Probióticos/isolamento & purificação , Rúmen/microbiologia , Leveduras/classificação , Ração Animal , Animais , Bovinos , Feminino , Fermentação , Rúmen/metabolismo , Leveduras/isolamento & purificação , Leveduras/fisiologiaRESUMO
The objective of this study was to evaluate 2 systems for covering corn silage in bunker silos. The first system consisted of a sheet of 45-µm-thick oxygen barrier film (OB, polyethylene + ethylene-vinyl alcohol) placed along the length of the sidewall before filling. After filling, the excess film was pulled over the wall on top of the silage, and a sheet of polyethylene was placed on top. The second system involved using a standard sheet (ST) of 180-µm-thick polyethylene film. Eight commercial bunker silos were divided into 2 parts lengthwise so that one-half of the silo was covered with OB and the other half with a ST system. During the filling, 3 net bags with chopped corn were buried in the central part (halfway between the top and bottom of the silo) of the bunkers (CCOR) in 3 sections 10 m apart. After filling, 18 net bags (9 per covering system) were buried 40 cm below the top surface of the 3 sections. These bags were placed at 3 distances from the bunker walls (0 to 50 cm, 51 to 100 cm, and 101 to 150 cm). During unloading, the bags were removed from the silos to determine the dry matter (DM) losses, fermentation end products, and nutritive value. The Milk2006 spreadsheet was used to estimate milk per tonne of DM. The model included the fixed effect of treatment (7 different locations in the bunker) and the random effect of the silo. Two contrasts were tested to compare silages in the top laterals (shoulders) with that in the CCOR (CCOR vs. OB and CCOR vs. ST). Three contrasts compared the corresponding distances of the silage covered by the 2 systems (OB50 vs. ST50, OB100 vs. ST100 and OB150 vs. ST150). Variables were analyzed with the PROC MIXED procedure of the SAS at the 5% level. The OB method produced well-fermented silages, which were similar to CCOR, whereas the OB system showed less lactic acid and greater pH and mold counts compared with CCOR. The ST method had 116.2 kg of milk/t less than the CCOR, as the OB system and the CCOR were similar (1,258.3 and 1,294.0 kg/t, respectively). Regarding the distances from the walls, the effects were more pronounced from 0 to 101 cm. The OB50 and OB100 silages had better quality and lower mold counts and DM losses than ST50 and ST100. The OB system reduced DM and nutrient losses at the shoulders in farm bunker corn silages compared with no sidewall plastic. The OB film should lap onto the crop for at least 200 cm so that 150 cm are covered outward from the wall.
Assuntos
Fermentação , Armazenamento de Alimentos/métodos , Valor Nutritivo , Oxigênio , Plásticos/química , Silagem , Zea mays , Criação de Animais Domésticos , Animais , Etanol , Armazenamento de Alimentos/instrumentação , Concentração de Íons de Hidrogênio , Ácido Láctico/análise , Polietileno , Compostos de VinilaRESUMO
AIMS: The aim of this study was to evaluate the chemical and microbiological characteristics and to identify the lactic acid bacteria (LAB) and yeasts involved in rehydrated corn kernel silage. METHODS AND RESULTS: Four replicates for each fermentation time: 5, 15, 30, 60, 90, 150, 210 and 280 days were prepared. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and PCR-based identification were utilized to identify LAB and yeasts. Eighteen bacteria and four yeast species were identified. The bacteria population reached maximum growth after 15 days and moulds were detected up to this time. The highest dry matter (DM) loss was 7·6% after 280 days. The low concentration of water-soluble carbohydrates (20 g kg-1 of DM) was not limiting for fermentation, although the reduction in pH and acid production occurred slowly. Storage of the rehydrated corn kernel silage increased digestibility up to day 280. CONCLUSIONS: This silage was dominated by LAB but showed a slow decrease in pH values. This technique of corn storage on farms increased the DM digestibility. SIGNIFICANCE AND IMPACT OF THE STUDY: This study was the first to evaluate the rehydrated corn kernel silage fermentation dynamics and our findings are relevant to optimization of this silage fermentation.
Assuntos
Bactérias/classificação , Fermentação , Silagem/microbiologia , Bactérias/isolamento & purificação , Bactérias/metabolismo , Ácido Láctico/metabolismo , Reação em Cadeia da Polimerase , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Leveduras/metabolismo , Zea mays/microbiologiaRESUMO
Accurate and rapid determination of trypanosomatids is essential in epidemiological surveillance and therapeutic studies. Matrix-assisted laser desorption ionization/time of flight mass spectrometry (MALDI-TOF MS) has been shown to be a useful and powerful technique to identify bacteria, fungi, metazoa and human intact cells with applications in clinical settings. Here, we developed and optimized a MALDI-TOF MS method to profile trypanosomatids. trypanosomatid cells were deposited on a MALDI target plate followed by addition of matrix solution. The plate was then subjected to MALDI-TOF MS measurement to create reference mass spectra library and unknown samples were identified by pattern matching using the BioTyper software tool. Several m/z peaks reproducibly and uniquely identified trypanosomatids species showing the potentials of direct identification of trypanosomatids by MALDI-TOF MS. Moreover, this method discriminated different life stages of Trypanosoma cruzi, epimastigote and bloodstream trypomastigote and Trypanosoma brucei, procyclic and bloodstream. T. cruzi Discrete Typing Units (DTUs) were also discriminated in three clades. However, it was not possible to achieve enough resolution and software-assisted identification at the strain level. Overall, this study shows the importance of MALDI-TOF MS for the direct identification of trypanosomatids and opens new avenues for mass spectrometry-based detection of parasites in biofluids. Copyright © 2016 John Wiley & Sons, Ltd.
Assuntos
Parasitologia/métodos , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos , Trypanosoma/química , Trypanosoma/isolamento & purificação , Animais , Linhagem Celular , Haplorrinos , Humanos , Microscopia , Tripanossomíase/parasitologiaRESUMO
Forty-two enrofloxacin-resistant Escherichia coli strains isolated from eggs and first-week mortality associated with yolk sac infection of two vertically integrated poultry companies of Central Mexico in 1997 and 2005 were characterised. E. coli resistance to 19 antibiotics was determined, as well as the minimum inhibitory concentrations (broth dilution) for ciprofloxacin. The presence of gyrA,B, parC,E chromosomal point mutations, qnrA,B,S plasmid genes and the aminoglycoside acetyltransferase aac(6')-Ib-cr were determined by PCR and sequencing. Resistance to ampicillin (95%), piperacillin (95%), gatifloxacin (95%), levofloxacin (95%), ampicillin/sulbactam (90%), cefazolin (85%), trimethoprim/sulfamethoxazole (80%), amoxicillin/clavulanic acid (80%), aztreonam (80%), cefepime (80%), cefotaxime (80%), ceftazidime (80%), ceftriaxone (80%) and cefoxitin (75%) was high in the 2005 strains and 19 (95%) strains were resistant to 7 or more antimicrobials. The strains from 1997 expressed high rates of resistance only to the fluoroquinolones and 4 strains (18%) expressed resistance to 7 or more antimicrobials. All strains had a gyrA mutation (Ser83Leu) and a parC mutation (Ser80Ile or Ser80Arg) and 41 (97.6%) strains had a second gyrA mutation (Asp87Asn, Asp87Tyr or Asp87Gly). Only two (4.7%) strains had a parE mutation (Ser458Ala). A total of 10 strains were positive for the aac(6')-Ib wild-type gene, 6 strains for the aac(6')-Ib-cr variant and 6 strains possessed both the wild type and the variant. No gyrB mutations or qnrA,B,S genes were detected. This is the first report in Latin America of chromosomal and plasmid quinolone resistance genes in E. coli strains recovered from poultry.