scholarly journals Flow Cytometry to Assess the Counts and Physiological State of Cronobacter sakazakii Cells after Heat Exposure

Foods ◽  
2019 ◽  
Vol 8 (12) ◽  
pp. 688 ◽  
Author(s):  
Paloma Cal-Sabater ◽  
Irma Caro ◽  
María J. Castro ◽  
María J. Cao ◽  
Javier Mateo ◽  
...  

Cronobacter sakazakii is an opportunistic pathogen that is associated with outbreaks of neonatal necrotizing enterocolitis, septicaemia, and meningitis. Reconstituted powdered infant formulae is the most common vehicle of infection. The aim of the present study is to gain insight into the physiological states of C. sakazakii cells using flow cytometry to detect the compromised cells, which are viable but non-culturable using plate-based methods, and to evaluate the impact of milk heat treatments on those populations. Dead-cell suspensions as well as heat-treated and non-heat-treated cell suspensions were used. After 60 or 65 °C treatments, the number of compromised cells increased as a result of cells with compromised membranes shifting from the heat-treated suspension. These temperatures were not effective at killing all bacteria but were effective at compromising their membranes. Thus, mild heat treatments are not enough to guarantee the safety of powered infant formulae. Flow cytometry was capable of detecting C. sakazakii’s compromised cells that cannot be detected with classical plate count methods; thus, it could be used as a screening test to decrease the risk derived from the presence of pathogenic viable but non-culturable cells in this food that is intended for newborns’ nutrition.

Antibiotics ◽  
2021 ◽  
Vol 10 (12) ◽  
pp. 1440
Author(s):  
Christian James ◽  
Ronald Dixon ◽  
Luke Talbot ◽  
Stephen J. James ◽  
Nicola Williams ◽  
...  

The dissemination of antibiotic resistance genes (ARGs) is a global health concern. This study identifies and critically reviews the published evidence on whether cooking (heating) food to eliminate bacterial contamination induces sufficient damage to the functionality of ARGs. Overall, the review found that there is evidence in the literature that Antimicrobial Resistant (AMR) bacteria are no more heat resistant than non-AMR bacteria. Consequently, recommended heat treatments sufficient to kill non-AMR bacteria in food (70 °C for at least 2 min, or equivalent) should be equally effective in killing AMR bacteria. The literature shows there are several mechanisms through which functional genes from AMR bacteria could theoretically persist in heat-treated food and be transferred to other bacteria. The literature search found sparce published evidence on whether ARGs may actually persist in food after effective heat treatments, and whether functional genes can be transferred to other bacteria. However, three publications have demonstrated that functional ARGs in plasmids may be capable of persisting in foods after effective heat treatments. Given the global impact of AMR, there is clearly a need for further practical research on this topic to provide sufficient evidence to fully assess whether there is a risk to human health from the persistence of functional ARGs in heat-treated and cooked foods.


1982 ◽  
Vol 45 (6) ◽  
pp. 513-515 ◽  
Author(s):  
G. F. SENYK ◽  
R. R. ZALL ◽  
W. F. SHIPE

Raw milk was heat-treated under subpasteurization and suprapasteurization conditions, cooled and stored for up to 72 h at 4.4 and 6.7°C. Milk lipase activity and bacteria counts were monitored in both unheated and heated milks. Inhibition of milk lipase activity ranged from 42 to 98% for treatments of 57.2°C for 10 sec to 73.9°C for 10 sec, respectively. The logs of Standard Plate Count after 72 h of storage at 6.7°C were 6.56, 4.86, 4.31, 4.00 and 2.82 for unheated and 10-sec heat treatments at 57.2, 65.6, 73.9 and 82.2°C, respectively. Psychrotrophic Bacteria Counts were also lower in the heated milks than in the unheated milk. The logs of Psychrotrophic Bacteria Counts after 72 h of storage at 6.7°C were 6.21, 2.45, 2.27, 1.33 and 1.00 for unheated and 10-sec heat treatments at 57.2, 65.6, 73.9 and 82.2°C, respectively. Heat treatment of raw milk supplies would result in limiting action of the milk lipase system and growth of bacteria.


2000 ◽  
Vol 66 (3) ◽  
pp. 1228-1232 ◽  
Author(s):  
Thusitha S. Gunasekera ◽  
Paul V. Attfield ◽  
Duncan A. Veal

ABSTRACT Application of flow cytometry (FCM) to microbial analysis of milk is hampered by the presence of milk proteins and lipid particles. Here we report on the development of a rapid (≤1-h) FCM assay based on enzymatic clearing of milk to determine total bacteria in milk. When bacteria were added to ultra-heat-treated milk, a good correlation (r ≥ 0.98) between the FCM assay and the more conventional methods of plating and direct microscopic counting was achieved. Raw milk data showed a significant correlation (P < 0.01) and a good agreement (r = 0.91) between FCM and standard plate count methods. The detection limit of the FCM assay was ≤104 bacteria ml of milk−1. This limit is below the level of detection required to satisfy legislation in many countries and states.


2002 ◽  
Vol 92 (9) ◽  
pp. 956-963 ◽  
Author(s):  
Menachem P. Assaraf ◽  
Chen Ginzburg ◽  
Jaacov Katan

Survival of Fusarium oxysporum f. sp. niveum following heat treatments was studied using flow cytometric, physiological, and microscopic assays. We exposed germinating conidia to sublethal temperatures from 36 to 42°C for 60 min, followed by rhodamine 123 staining and flow cytometry, and found increasing levels of fluorescence that reflect a change in mitochondrial membrane potential, indicating a weakening induced by stress. Viability of conidia or germinating conidia of the fungus exposed to heat decreased with increasing temperature, as assessed by fluorescent staining. However, viability was higher than that assessed with the 5-day-long plate count method and was further reduced 13 and 24 h after treatment, suggesting delayed mortality of the heat-treated germinating conidia. Delayed mortality was substantiated by observing these conidia with light and fluorescent scanning electron microscopy and by subculturing single germinating conidia that had been previously heated. Programmed cell death was not observed in heat-treated conidia or germinating conidia of F. oxysporum based on the detection of plasma membrane phosphatidylserine translocation, cell-cycle measurements, detection of DNA fragmentation, or microscopic observation of apoptotic bodies. We hypothesize that propagules, which survived the heating and apparently are alive, may undergo further irreversible detrimental processes, eventually leading to their death by yet unidentified mechanisms. These findings suggest that pathogen propagules also might be affected under lower temperatures, possibly facilitating pathogen control by heating.


2015 ◽  
Vol 2 (1) ◽  
pp. 30-34
Author(s):  
K. Korobkova ◽  
V. Patyka

Contemporary state of the distribution of mycoplasma diseases of cultivated crops in Ukraine was analyzed. The changes of the physiological state of plant cells under the impact of mollicutes were investigated. It was demonstrated that there is temporary increase in the activity of peroxidase, catalase, polyphenoloxidase, phenylalanine-ammonia-lyase at the early stages of interaction. The adhesive properties are changed in the mollicutes under the impact of plant lectin; there is synthesis of new polypeptides. It was determined that the phytopathogenic acholeplasma is capable of producing a complex of proteolytic enzymes into the culture me- dium. It was concluded that when plant cells are infected with acholeplasma, a number of signaling interactions and metabolic transformations condition the recognition of pathogenesis and ensure the aggregate response of a plant to stress in the form of defense reactions. It was assumed that some specifi cities of the biology of phy- topathogenic acholeplasma determine their avoiding the immune mechanisms of plants and promote long-term persistence of mollicutes.


Author(s):  
G.A. Murachueva ◽  
I.M. Rasulov ◽  
S.G. Gusenov

A review of the literature on the stages of the formation of temporary and permanent occlusion has been performed. This stages play an important role not only for the full development of the maxillofacial apparatus, temporomandibular joint, but also the whole organism. The role of early tooth extraction in the formation of the physiological state of the dentoalveolar system is considered. The conclusion is drawn about the need for a deeper study of this problem in the structure of general dental morbidity.


Domiati cheese is the most popular brand of cheese ripened in brine in the Middle East in terms of consumed quantities. This study was performed to investigate the impact of the microbiological quality of the used raw materials, the applied traditional processing techniques and ripening period on the quality and safety of the produced cheese. Three hundred random composite samples were collected from three factories at Fayoum Governorate, Egypt. Collected samples represent twenty-five each of: raw milk, table salt, calf rennet, microbial rennet, water, environmental air, whey, fresh cheese, ripened cheese & swabs from: worker hands; cheese molds and utensils; tanks. All samples were examined microbiologically for Standard Plate Count (SPC), coliforms count, Staphylococcus aureus (S. aureus) count, total yeast & mould count, presence of E. coli, Salmonellae and Listeria monocytogenes (L. monocytogenes). The mean value of SPC, coliforms, S. aureus and total yeast & mould counts ranged from (79×102 CFU/m3 for air to 13×108 CFU/g for fresh cheese), (7×102 MPN/ cm2 for tank swabs to 80×106 MPN/ml for raw milk), (9×102 CFU/g for salt to 69×106 CFU/g for fresh cheese) and (2×102 CFU/cm2 for hand swabs to 60×104 CFU/g for fresh cheese), respectively. Whereas, E. coli, Salmonella and L. monocytogenes failed to be detected in all examined samples. There were significant differences in all determined microbiological parameters (p ≤0.05) between fresh and ripened cheese which may be attributed to different adverse conditions such as water activity, pH, salt content and temperature carried out to improve the quality of the product.


Author(s):  
Marios Kazasidis ◽  
Elisa Verna ◽  
Shuo Yin ◽  
Rocco Lupoi

AbstractThis study elucidates the performance of cold-sprayed tungsten carbide-nickel coating against solid particle impingement erosion using alumina (corundum) particles. After the coating fabrication, part of the specimens followed two different annealing heat treatment cycles with peak temperatures of 600 °C and 800 °C. The coatings were examined in terms of microstructure in the as-sprayed (AS) and the two heat-treated conditions (HT1, HT2). Subsequently, the erosion tests were carried out using design of experiments with two control factors and two replicate measurements in each case. The effect of the heat treatment on the mass loss of the coatings was investigated at the three levels (AS, HT1, HT2), as well as the impact angle of the erodents (30°, 60°, 90°). Finally, the response surface methodology (RSM) was applied to analyze and optimize the results, building the mathematical models that relate the significant variables and their interactions to the output response (mass loss) for each coating condition. The obtained results demonstrated that erosion minimization was achieved when the coating was heat treated at 600 °C and the angle was 90°.


2021 ◽  
Vol 21 (3) ◽  
Author(s):  
Przemysław Snopiński ◽  
Mariusz Król ◽  
Marek Pagáč ◽  
Jana Petrů ◽  
Jiří Hajnyš ◽  
...  

AbstractThis study investigated the impact of the equal channel angular pressing (ECAP) combined with heat treatments on the microstructure and mechanical properties of AlSi10Mg alloys fabricated via selective laser melting (SLM) and gravity casting. Special attention was directed towards determining the effect of post-fabrication heat treatments on the microstructural evolution of AlSi10Mg alloy fabricated using two different routes. Three initial alloy conditions were considered prior to ECAP deformation: (1) as-cast in solution treated (T4) condition, (2) SLM in T4 condition, (3) SLM subjected to low-temperature annealing. Light microscopy, transmission electron microscopy, X-ray diffraction line broadening analysis, and electron backscattered diffraction analysis were used to characterize the microstructures before and after ECAP. The results indicated that SLM followed by low-temperature annealing led to superior mechanical properties, relative to the two other conditions. Microscopic analyses revealed that the partial-cellular structure contributed to strong work hardening. This behavior enhanced the material’s strength because of the enhanced accumulation of geometrically necessary dislocations during ECAP deformation.


Author(s):  
A. Brown ◽  
K. Krishnan ◽  
L. Wayne ◽  
P. Peralta ◽  
S. N. Luo ◽  
...  

Global and local microstructural weak links for spall damage were investigated using 3-D characterization in polycrystalline (PC) and multicrystalline (MC) copper samples, respectively. All samples were shocked via flyer-target plate experiments using a laser drive at low pressures (2–6 GPa). The flyer plates measured approximately 500 μm thick and 8 mm in diameter and the target plates measured approximately 1000 μm thick and 10 mm in diameter. Electron Backscattering Diffraction (EBSD) and optical microscopy were used to determine to presence of voids and relate them to the surrounding microstructure. Statistics on the strength of grain boundaries (GBs) was conducted by analyzing PC samples and collecting the misorientation across GBs with damage present, and it was found that a misorientation range of 25–50° is favorable for damage. Statistics were also taken of copper PC samples that had undergone different heat treatments and it was found that although the 25–50° range is less dominant, it is still favorable for damage nucleation. Removal of initial plastic strain via heat treatments and an increase in Σ3 CSL boundaries, indicative of strong annealing twins, also led to an increased amount of transgranular damage. 3-D X-ray tomography data were used to investigate the shape of the voids present in untreated, as received and heat treated samples. It was found that the as received sample contained a higher amount of “disk”, or, “sheet-like” voids indicative of intergranular damage, whereas the heat treated samples had a higher fraction of spherical shaped voids, indicative of transgranular damage. MC samples were used to study microstructural weak links for spall damage because the overall grain size is much larger than the average void size, making it possible to determine which GBs nucleated damage. Simulations and experimental analysis of damage sites with large volumes indicate that high Taylor factor mismatches with respect to the crystallographic grain GB normal is the primary cause for the nucleation of damage at a GB interface and a low Taylor factor along the shock direction in either grain drives void growth perpendicular to the GB. Cases where experimental results show damage and simulation results show no damage are attributed to the presence of an intrinsic microstructural weak link, such as an incoherent twin boundary.


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