Monitoramento da distribuição do tamanho das partículas do leite integral e desnatado durante os processos de coagulação ácida ou enzimática

O processo de coagulação do leite pode ocorrer, principalmente, por duas vias: coagulação ácida ou enzimática, sendo a característica do gel obtido dependente do tipo de coagulação, do tipo de processamento aplicado e da composição do leite inicial. Portanto, o objetivo deste trabalho foi avaliar as modificações microestruturais do leite integral e desnatado pasteurizados através da análise de distribuição de partículas (LS), mediante coagulações por via ácida, realizada via fermentação pela adição da cultura mista de Streptococcus thermophilus e Lactobacillus delbrueckii subsp. bulgaricus; e enzimática, por adição de quimosina microbiana. Durante todo o processo de coagulação, foram retiradas alíquotas de leite de ambas as coagulações até a obtenção dos géis. Os géis obtidos apresentaram diferenças no tempo de coagulação e na distribuição do tamanho das partículas, além disso, os tamanhos das partículas dos géis do leite desnatado apresentaram valores finais em todas as distribuições, maiores que o leite integral, possivelmente relacionados ao fato da caseína, proteína majoritária, estar mais disponível no meio reacional para que ocorra a desestabilização do sistema em ambos os tipos de coagulação. Ao comparar os dois processos de coagulação para a distribuição d90, os géis da coagulação enzimática apresentaram agregados 12 vezes maiores que os géis da coagulação ácida, isso demonstrando que as forças de interações para a formação dos agregados são muito maiores na coagulação enzimática.

Antimicrobial Susceptibility of Lactobacillus delbrueckii subsp. lactis from Milk Products and Other Habitats

As components of many cheese starter cultures, strains of Lactobacillus delbrueckii subsp. lactis (LDL) must be tested for their antimicrobial susceptibility to avoid the potential horizontal transfer of antibiotic resistance (ABR) determinants in the human body or in the environment. To this end, a phenotypic test, as well as a screening for antibiotic resistance genes (ARGs) in genome sequences, is commonly performed. Historically, microbiological cutoffs (MCs), which are used to classify strains as either ‘sensitive’ or ‘resistant’ based on the minimal inhibitory concentrations (MICs) of a range of clinically-relevant antibiotics, have been defined for the whole group of the obligate homofermentative lactobacilli, which includes LDL among many other species. This often leads to inaccuracies in the appreciation of the ABR status of tested LDL strains and to false positive results. To define more accurate MCs for LDL, we analyzed the MIC profiles of strains originating from various habitats by using the broth microdilution method. These strains’ genomes were sequenced and used to complement our analysis involving a search for ARGs, as well as to assess the phylogenetic proximity between strains. Of LDL strains, 52.1% displayed MICs that were higher than the defined MCs for kanamycin, 9.9% for chloramphenicol, and 5.6% for tetracycline, but no ARG was conclusively detected. On the other hand, all strains displayed MICs below the defined MCs for ampicillin, gentamycin, erythromycin, and clindamycin. Considering our results, we propose the adaptation of the MCs for six of the tested clinically-relevant antibiotics to improve the accuracy of phenotypic antibiotic testing.

Optimization of compatible solutes for improving survival of freeze-dried Lactobacillus delbrueckii subsp. bulgaricus using Box-Behnken design

Lactobacillus delbrueckii subsp. bulgaricus is widely used in yogurt as a starter. The freeze-drying process may cause bacteria death. In the present work, the effect of three solutes (NaCl, sorbitol, and sodium glutamate) in MRS on viability of L.bulgaricus during freeze-drying was investigated. The optimal combination of adequate solutes was chosen by Box-Behnken Design. The survival rate and viable counts in freeze-dried powder, as well as the viable counts in broth, were used as responses. The results revealed that the optimum combination of solutes in MRS broth were 0.50% NaCl, 0.19% sorbitol, and 0.06% sodium glutamate. Under these optimal conditions, the survival rate was 53.2±0.14%, the viable counts in freeze-dried powder was 8.51±0.23×1010 CFU/g, and the viable counts in broth was 6.05±0.19 ×108 CFU/mL, which were increased by 17.18%, 15.94%, and 17.31%, respectively, compared to the control. This research demonstrated the possibility of viability improvement of L.bulgaricus, which may provide a feasible reference for industrial development.

Oleate Hydratase in Lactobacillus delbrueckii subsp. bulgaricus LBP UFSC 2230 Catalyzes the Reversible Conversion between Linoleic Acid and Ricinoleic Acid

This study provides insight into the enzymatic mechanism of CLA synthesis in L. delbrueckii subsp. bulgaricus and broadens our understanding of the bioconversion of LA and RA by OleH. The impact of OleH on the production of the c 9, t 11 CLA isomer and stress tolerance by E. coli has been assisted.

Isolation of Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus from nature: Technological characterisation and antibiotic resistance

Yoghurt fermenting bacteria were isolated from natural sources including plants, dew, and rain samples (total of 300 samples) by the same methods nomadic peoples used for several centuries in Turkey. Inoculation into the reconstituted skim milk followed by planting on specific media and matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry (MALDI-TOF MS) analysis allowed for the identification of 18 Lactobacillus delbrueckii subsp. and 26 Streptococcus thermophilus. A multiplex polymerase chain reaction (PCR) assay applied to lactobacilli enabled the identification of 5 isolates as L. delbrueckii subsp. bulgaricus. The isolates showed a varying range of acidification rates and proteolytic activity in reconstituted skimmed milk (RSM). S. thermophilus isolates showed a broader range of resistance and the most frequent resistance was observed for streptomycin (69.2%), gentamycin (65.3%), clindamycin (61.5%), ampicillin (61.5%), kanamycin (53.8%), and erythromycin (50%). For L. delbrueckii subsp. the highest resistance was determined for vancomycin (38.8%), ciprofloxacin (33.3%), and penicillin (27.8%). The frequency of multiple resistance was tested on 14 different antimicrobials determining that 19 S. thermophilus (73%) and 3 L. delbrueckii subsp. (16.7%) demonstrated resistance to more than three different antibiotics. In contrast to this wide-ranging resistance, five isolates from each genus were found to be susceptible to all tested antibiotics. The present study indicates that lactic acid bacteria (LAB) isolated from nature may have broad-range of resistance to antibiotics and could be a source for the transfer of resistance.