Susceptibility of Bifidobacteria of Animal Origin to Selected Antimicrobial Agents

Strains of the genus Bifidobacterium are frequently used as probiotics, for which the absence of acquired antimicrobial resistance has become an important safety criterion. This clarifies the need for antibiotic susceptibility data for bifidobacteria. Based on a recently published standard for antimicrobial susceptibility testing of bifidobacteria with broth microdilution method, the range of susceptibility to selected antibiotics in 117 animal bifidobacterial strains was examined. Narrow unimodal MIC distributions either situated at the low-end (chloramphenicol, linezolid, and quinupristin/dalfopristin) or high-end (kanamycin, neomycin) concentration range could be detected. In contrast, the MIC distribution of trimethoprim was multimodal. Data derived from this study can be used as a basis for reviewing or verifying present microbiological breakpoints suggested by regulatory agencies to assess the safety of these micro-organisms intended for the use in probiotics

Uporaba lokalnih sojeva bakterije Lactococcus lactis kao starter kultura u proizvodnji tradicionalnih crnogorskih sireva

The aim of this study is to characterise and examine the biochemical properties of 40 Lactococcus lactis strains isolated from indigenous Montenegrin dairy products in order to explore their potential to be used as starter cultures for producing typical Montenegrin cheese, such as ‘bijeli sir’, ‘masni sir’ and ‘njeguški sir’. Their safety regarding the production of biogenic amines, the presence of antimicrobial resistance and the antibacterial activity against relevant pathogens and spoilage microorganisms has also been tested. Based on the characterisation, all strains belong to L. lactis ssp. lactis. Out of these 40 strains, 23 displayed rapid acidification ability and proteolysis. However, none of the strains exhibited the ability of lipid degradation. Most of the strains were not associated with any health risk investigated. Summing up, a large percentage (27.5 %) of the tested strains showed good properties. These strains should be further examined for their possible application as specific starter cultures in the production of indigenous cheese in MontenegroSvrha je ovoga rada bila okarakterizirati i ispitati biokemijska svojstva 40 sojeva bakterije Lactococcus lactis, izoliranih iz autohtonih crnogorskih mliječnih proizvoda, te istražiti mogućnost njihove uporabe kao starter kultura u proizvodnji tipičnih crnogorskih sireva, kao što su bijeli sir, masni sir i njeguški sir. Također je ispitana sigurnost primjene tih sojeva, tj. sposobnost proizvodnje biogenih amina, antimikrobna rezistencija i sposobnost suzbijanja rasta važnih patogenih mikroorganizama i uzročnika kvarenja hrane. Karakterizacijom je utvrđeno da svih 40 sojeva pripadaju podvrsti L. lactis ssp. lactis. Njih 23 imala su sposobnost brzog zakiseljavanja i proteolize. Međutim, niti jedan soj nije pokazao sposobnost razgradnje lipida. Većina sojeva nije predstavljala nikakav rizik za zdravlje. Velik postotak sojeva (27,5 %) pokazao je dobra proizvodna svojstva, pa treba dodatno ispitati mogućnost njihove uporabe kao specifičnih starter kultura u proizvodnji autohtonih crnogorskih sireva

Ropiness in Bread—A Re-Emerging Spoilage Phenomenon

As bread is a very important staple food, its spoilage threatens global food security. Ropy bread spoilage manifests in sticky and stringy degradation of the crumb, slime formation, discoloration, and an odor reminiscent of rotting fruit. Increasing consumer demand for preservative-free products and global warming may increase the occurrence of ropy spoilage. Bacillus amyloliquefaciens, B. subtilis, B. licheniformis, the B. cereus group, B. pumilus, B. sonorensis, Cytobacillus firmus, Niallia circulans, Paenibacillus polymyxa, and Priestia megaterium were reported to cause ropiness in bread. Process hygiene does not prevent ropy spoilage, as contamination of flour with these Bacillus species is unavoidable due to their occurrence as a part of the endophytic commensal microbiota of wheat and the formation of heat-stable endospores that are not inactivated during processing, baking, or storage. To date, the underlying mechanisms behind ropy bread spoilage remain unclear, high-throughput screening tools to identify rope-forming bacteria are missing, and only a limited number of strategies to reduce rope spoilage were described. This review provides a current overview on (i) routes of entry of Bacillus endospores into bread, (ii) bacterial species implicated in rope spoilage, (iii) factors influencing rope development, and (iv) methods used to assess bacterial rope-forming potential. Finally, we pinpoint key gaps in knowledge and related challenges, as well as future research questions

Clostridium strain FAM25158, a unique endospore-forming bacterium related to Clostridium tyrobutyricum and isolated from Emmental cheese shows low tolerance to salt

The genus Clostridium is a large and diverse group of species that can cause food spoilage, including late blowing defect (LBD) in cheese. In this study, we investigated the taxonomic status of strain FAM25158 isolated from Emmental cheese with LBD using a polyphasic taxonomic and comparative genomic approach. A 16S rRNA gene sequence phylogeny suggested affiliation to the Clostridium sensu stricto cluster, with Clostridium tyrobutyricum DSM 2637T being the closest related type strain (99.16% sequence similarity). Average Nucleotide Identity (ANI) analysis revealed that strain FAM25158 is at the species threshold with C. tyrobutyricum, with ANI values ranging from 94.70 to 95.26%, while the digital DNA-DNA hybridization values were below the recommended threshold, suggesting that FAM25158 is significantly different from C. tyrobutyricum at the genomic level. Moreover, comparative genomic analysis between FAM25158 and its four closest C. tyrobutyricum relatives revealed a diversity of metabolic pathways, with FAM25158 differing from other C. tyrobutyricum strains by the presence of genes such as scrA, srcB, and scrK, responsible for sucrose utilization, and the absence of many important functional genes associated with cold and osmolality adaptation, which was further supported by phenotypic analyses. Surprisingly, strain FAM25158 exhibited unique physiologic traits, such as an optimal growth temperature of 30°C, in contrast to its closest relatives, C. tyrobutyricum species with an optimal growth temperature of 37°C. Additionally, the growth of FAM25158 was inhibited at NaCl concentrations higher than 0.5%, a remarkable observation considering its origin from cheese. While the results of this study provide novel information on the genetic content of strain FAM25158, the relationship between its genetic content and the observed phenotype remains a topic requiring further investigation

Ropiness in bread : a re-emerging spoilage phenomenon

As bread is a very important staple food, its spoilage threatens global food security. Ropy bread spoilage manifests in sticky and stringy degradation of the crumb, slime formation, discoloration, and an odor reminiscent of rotting fruit. Increasing consumer demand for preservative-free products and global warming may increase the occurrence of ropy spoilage. Bacillus amyloliquefaciens, B. subtilis, B. licheniformis, the B. cereus group, B. pumilus, B. sonorensis, Cytobacillus firmus, Niallia circulans, Paenibacillus polymyxa, and Priestia megaterium were reported to cause ropiness in bread. Process hygiene does not prevent ropy spoilage, as contamination of flour with these Bacillus species is unavoidable due to their occurrence as a part of the endophytic commensal microbiota of wheat and the formation of heat-stable endospores that are not inactivated during processing, baking, or storage. To date, the underlying mechanisms behind ropy bread spoilage remain unclear, high-throughput screening tools to identify rope-forming bacteria are missing, and only a limited number of strategies to reduce rope spoilage were described. This review provides a current overview on (i) routes of entry of Bacillus endospores into bread, (ii) bacterial species implicated in rope spoilage, (iii) factors influencing rope development, and (iv) methods used to assess bacterial rope-forming potential. Finally, we pinpoint key gaps in knowledge and related challenges, as well as future research questions

The Influence of Meat Batter Composition and Sausage Diameter on Microbiota and Sensory Traits of Artisanal Wild Boar Meat Sausages

U ovom radu istražen je utjecaj sastava mesnog nadjeva i promjera kobasica na razvoj mikrobiote i senzornih osobina tradicijskih, spontano fermentiranih kobasica od mesa divlje svinje. Ovo istraživanje pokazuje kako se analizom glavnih komponenata mogu povezati senzorna svojstva proizvoda s određenim sojem bakterija, te za selekciju potencijalnih starter kultura. Općenito govoreći, nakon mikrobioloških analiza svih vrsta proizvedenih kobasica dobiveni su slični rezultati. Niti u jednoj fazi proizvodnje kobasica nije detektirana neželjena mikrobiota ili je njezin broj u zrelim kobasicama bio ispod granice detekcije. Niska stopa rasta bakterija mliječne kiseline bila je u skladu s dobivenim pH-vrijednostima i sporom stopom zakiseljavanja. Iako nisu uočene razlike u sastavu vrsta bakterija mliječne kiseline između različitih tipova kobasica (50S=50 % mesa divlje svinje u tankom ovitku, 50L=50 % mesa divlje svinje u debelom ovitku, 100S= 100 % mesa divlje svinje u tankom ovitku), moglo se opaziti jasno odvajanje na osnovi genotipova pojedinih bakterija. Kvantitativnom opisnom analizom utvrđene su bitne razlike u senzornim značajkama različitih vrsta kobasica. Prema analizi glavnih komponenata, svojstva dopadljivosti kobasica usko su povezana s jednim genotipom Leuconostoc mesenteroides (LM_4). Od svih ispitanih tehnoloških svojstava, sojevi LM_4 pokazali su izuzetnu sposobnost zakiseljavanja, snižavajući pH-vrijednost s 5,41 na 3,74, te izrazito proteolitičko djelovanje u obranom mlijeku, te antagonističku aktivnost prema bakterijama Staphylococcus aureus (DSM 20231) i Brochothrix thermosphacta (LMG 17208). Lipolitička i hemolitička aktivnost nije detektirana, a svi analizirani sojevi bili su osjetljivi na testirane antibiotike i nisu posjedovali gene za biogene amine.In this study, the influence of meat batter composition and sausage diameter on the development of microbiota and sensory traits of traditional, spontaneously fermented wild boar meat sausages are evaluated. This research also demonstrates how principal component analysis (PCA) can be used to relate product sensory properties to particular microbial genotype and to select potential starter or adjunct culture. Generally, similar microbiological results were obtained in all types of products. The undesirable microbiota was either not detected at any sausage production stage or its number decreased below the detection limit in ripened sausages. The low growth rate of lactic acid bacteria (LAB) was consistent with the obtained pH and slow acidification rate. Although no differences in the composition of LAB species were noticed between sausage types (50S=50 % wild boar meat in small casing, 50L=50 % wild boar meat in large casing, 100S=100 % wild boar meat in small casing), a clear separation based on LAB genotypes could be observed. Upon quantitative descriptive analysis, significant differences in sensory attributes between sausage types were established. According to the PCA, the overall acceptability traits of sausages are closely linked to one Leuconostoc mesenteroides genotype (LM_4). Of all tested technological properties, LM_4 strains showed remarkable acidification ability, lowering the pH from pH=5.41 to 3.74, and pronounced proteolytic activity on skimmed milk as well as antagonistic activity against Staphylococcus aureus (DSM 20231) and Brochothrix thermosphacta (LMG 17208). Lipolytic and haemolytic activities were not detected, and all analyzed strains were susceptible to tested antibiotics and possessed no biogenic amine genes

Functional Properties and Sustainability Improvement of Sourdough Bread by Lactic Acid Bacteria

Preventing food spoilage without the addition of chemical food additives, while increasing functional properties of wheat-based bakery products, is an increasing demand by the consumers and a challenge for the food industry. Within this study, lactic acid bacteria (LAB) isolated from sourdough were screened in vitro for the ability to utilize the typical wheat carbohydrates, for their antimicrobial and functional properties. The dual culture overlay assay revealed varying levels of inhibition against the examined fungi, with Lactiplantibacillus plantarum S4.2 and Lentilactobacillusparabuchneri S2.9 exhibiting the highest suppression against the indicator strains Fusarium graminearum MUCL43764, Aspergillus fumigatus, A. flavus MUCL11945, A. brasiliensis DSM1988, and Penicillium roqueforti DSM1079. Furthermore, the antifungal activity was shown to be attributed mainly to the activity of acids produced by LAB. The antibacillus activity was evaluated by the spot-on-the-lawn method revealing a high inhibition potential of the majority of LAB isolated from sourdough against Bacillus cereus DSM31, B. licheniformis DSM13, B. subtilis LMG7135, and B. subtilis S15.20. Furthermore, evaluating the presence of the glutamate decarboxylase gen in LAB isolates by means of PCR showed a strain dependency of a potential GABA production. Finally, due to improved functional activities, LAB isolated from sourdoughs exhibit promising characteristics for the application as natural preservatives in wheat-based bakery products

Applicability of Yeast Fermentation to Reduce Fructans and Other FODMAPs

A diet low in fermentable oligosaccharides, disaccharides, monosaccharides and, polyols (FODMAPs) is recommended for people affected by irritable bowel syndrome (IBS) and non-coeliac wheat sensitivity (NCWS) in order to reduce symptoms. Therefore, the aim of this study was to evaluate the impact of 13 sourdough-related yeasts on FODMAP degradation, especially fructans. First, a model system containing a typical wheat carbohydrate profile was applied to evaluate the growth rate of each yeast strain. Additionally, changes in the sugar composition, for up to four days, were monitored by high-pressure anion-exchange chromatography (HPAEC). A more realistic approach with a wheat flour suspension was used to characterize CO2 production according to the Einhorn method. The reduction of the total fructans was analyzed using an enzymatic method. Furthermore, a fingerprint of the present fructans with different degrees of polymerization was analyzed by HPAEC. The results revealed strong differences in the examined yeast strains’ ability to degrade fructans, in both the model system and wheat flour. Overall, Saccharomyces cerevisiae isolated from Austrian traditional sourdough showed the highest degree of degradation of the total fructan content and the highest gas building capacity, followed by Torulaspora delbrueckii. Hence, this study provides novel knowledge about the FODMAP conversion of yeast strains

Immunogold Nanoparticles for Rapid Plasmonic Detection of C. sakazakii

Cronobacter sakazakii is a foodborne pathogen that can cause a rare, septicemia, life-threatening meningitis, and necrotizing enterocolitis in infants. In general, standard methods for pathogen detection rely on culture, plating, colony counting and polymerase chain reaction DNA-sequencing for identification, which are time, equipment and skill demanding. Recently, nanoparticle- and surface-based immunoassays have increasingly been explored for pathogen detection. We investigate the functionalization of gold nanoparticles optimized for irreversible and specific binding to C. sakazakii and their use for spectroscopic detection of the pathogen. We demonstrate how 40-nm gold nanoparticles grafted with a poly(ethylene glycol) brush and functionalized with polyclonal antibodies raised against C. sakazakii can be used to specifically target C. sakazakii. The strong extinction peak of the Au nanoparticle plasmon polariton resonance in the optical range is used as a label for detection of the pathogens. Individual binding of the nanoparticles to the C. sakazakii surface is also verified by transmission electron microscopy. We show that a high degree of surface functionalization with anti-C. sakazakii optimizes the detection and leads to a detection limit as low as 10 CFU/mL within 2 h using a simple cuvette-based UV-Vis spectrometric readout that has great potential for further optimization