A note on the use of the non-central t-distribution in setting numerical microbiological specifications for foods

O rdonez , J.A. S anz , B. H ernandez , P.E. L opez -L orenzo , P 1984, A note on the effect of combined ultrasonic and heat treatments on the survival of thermoduric streptococci. Journal of Applied Bacteriology , 56 , 175–177.
The combined destructive effect of ultrasonic waves and heating on microorganisms has been investigated using as a model two species of thermoduric streptococci. Ultrasonic and heat treatments applied simultaneously were much more destructive than the additive effect of the two agents considered independently. Compared with heat alone, the simultaneous application of ultrasonic vibration and heat reduced the resistance of the organisms 6–11.fold.     

Effect of heat and ultrasonic waves on the survival of two strains of Bacillus subtilis

The combined effect of ultrasonic (20 KHz, 150 W) and heat treatment on the survival of two strains of Bacillus subtilis in three suspending media (distilled water, glycerol and milk) has been studied. When spores suspended in water or milk were subjected to ultrasonic waves before heat treatments a little or no decrease of the heat resistance was observed. However, both sporicidal agents applied simultaneously (thermo-ultrasonication) decreased by 63% (B. subtilis, var. niger-40) and 74% (B. subtilis ATCC 6051) the decimal reduction times for the heat treatment when the spores were suspended in glycerol and by 79% and 40%, respectively when suspended in milk. The thermo-ultrasonication of spores in water markedly reduced the heat resistance of them (between 99.9% and 70%) in the range 70-95 degrees C but the effect of the thermo-ultrasonication significantly diminished as the temperature of the treatment was approached to the boiling point of the water.     

Ultrasound-biophysics mechanisms

Ultrasonic biophysics is the study of mechanisms responsible for how ultrasound and biological materials interact. Ultrasound-induced bioeffect or risk studies focus on issues related to the effects of ultrasound on biological materials. On the other hand, when biological materials affect the ultrasonic wave, this can be viewed as the basis for diagnostic ultrasound. Thus, an understanding of the interaction of ultrasound with tissue provides the scientific basis for image production and risk assessment. Relative to the bioeffect or risk studies, that is, the biophysical mechanisms by which ultrasound affects biological materials, ultrasound-induced bioeffects are generally separated into thermal and non-thermal mechanisms. Ultrasonic dosimetry is concerned with the quantitative determination of ultrasonic energy interaction with biological materials.

Whenever ultrasonic energy is propagated into an attenuating material such as tissue, the amplitude of the wave decreases with distance. This attenuation is due to either absorption or scattering. Absorption is a mechanism that represents that portion of ultrasonic wave that is converted into heat, and scattering can be thought of as that portion of the wave, which changes direction. Because the medium can absorb energy to produce heat, a temperature rise may occur as long as the rate of heat production is greater than the rate of heat removal. Current interest with thermally mediated ultrasound-induced bioeffects has focused on the thermal isoeffect concept. The non-thermal mechanism that has received the most attention is acoustically generated cavitation wherein ultrasonic energy by cavitation bubbles is concentrated. Acoustic cavitation, in a broad sense, refers to ultrasonically induced bubble activity occurring in a biological material that contains pre-existing gaseous inclusions. Cavitation-related mechanisms include radiation force, microstreaming, shock waves, free radicals, microjets and strain. It is more challenging to deduce the causes of mechanical effects in tissues that do not contain gas bodies. These ultrasonic biophysics mechanisms will be discussed in the context of diagnostic ultrasound exposure risk concerns.     

Factors contributing to heat resistance of Clostridium perfringens endospores

The endospores formed by strains of type A Clostridium perfringens that produce the C. perfringens enterotoxin (CPE) are known to be more resistant to heat and cold than strains that do not produce this toxin. The high heat resistance of these spores allows them to survive the cooking process, leading to a large number of food-poisoning cases each year. The relative importance of factors contributing to the establishment of heat resistance in this species is currently unknown. The present study examines the spores formed by both CPE(+) and CPE(-) strains for factors known to affect heat resistance in other species. We have found that the concentrations of DPA and metal ions, the size of the spore core, and the protoplast-to-sporoplast ratio are determining factors affecting heat resistance in these strains. While the overall thickness of the spore peptidoglycan was found to be consistent in all strains, the relative amounts of cortex and germ cell wall peptidoglycan also appear to play a role in the heat resistance of these strains.     

Synergism between Ultrasonic Waves and Hydrogen Peroxide in the Killing of Micro-organisms

The lethal effect on different micro-organisms of ultrasonic waves and hydrogen peroxide separately and in combination was examined. Ultrasonic waves were able to disintegrate Fusobacterium nucleatum within 3 min and to kill Veillonella parvula after 15 min and Streptoccus sanguis after 20 min; 20 vols H₂O₂ (6% w/v) killed V. parvula, Strep. sanguis and Staphylococcus aureus after 5 min treatment, and Clostridium sporogenes spores after 25 min. Sonication of Cl. sporogenes spores, Bacillus cereus spores and Candida albicans in 20 vols H₂O₂, using an ultrasonic probe, was lethal to the organisms after 15, 10 and 10 min, respectively. The latter 2 organisms were not killed by 30 min exposure to either agent separately. Similar results were obtained when an ultrasonic tank was used for sonication.     

Bacillus sporothermodurans and other highly heat-resistant spore formers in milk

A recent example of a micro-organism causing undesired growth in consumer milk is Bacillus sporothermodurans producing highly heat-resistant spores (HRS) which may survive ultra-high temperature (UHT) treatment or industrial sterilization. Molecular typing showed a heterogeneous group of farm isolates (non-HRS strains), but a clonal group of UHT isolates from diverse European countries and other continents (HRS-clone) suggesting a common source. During a survey of Belgian dairy farms for the presence of potentially highly heat-resistant spore formers, high numbers of these spores were detected in filter cloth, green crop and fodder samples. The strain collection showed a high taxonomic diversity with 18 potentially new species and with Bacillus licheniformis and Geobacillus pallidus as predominating species overall. Seventeen B. sporothermodurans isolates were identified, mainly originating from feed concentrate. Heat resistance studies showed the UHT resistance of B. sporothermodurans spores present in industrially contaminated UHT milk, but a lower heat resistance of laboratory-grown strains (HRS and non-HRS). Hydrogen peroxide, used as sanitizer in the dairy industry, was found to induce higher heat resistance of laboratory-grown B. sporothermodurans strains to a certain level. This indicates that sublethal stress conditions may affect the heat resistance. By transmission electron microscopy, structural differences at the spore level were found between HRS and non-HRS strains. The data indicate that the attainment of extreme heat resistance is rather multifactorial.     

Thermal resistance of wild-type and antibiotic-resistant Listeria monocytogenes in meat and potato substrates

AIMS: This study aimed to elucidate the relationship, if any, between the acquisition/possession of antibiotic resistance in strains of Listeria monocytogenes and the resistance of such strains to heat stress. METHODS AND RESULTS: D-values calculated using a linear survival model were used to compare the heat resistance of two wild-type (WT) and two antibiotic (streptomycin)-resistant (AR) mutant strains of L. monocytogenes measured in minced beef and potato substrates at 55 degrees C, with and without prior heat shock at 48 degrees C. In both minced beef and potato, no significant differences (P < 0.05) between D-values of AR and WT strains were noted. Heat shock did not significantly increase D-values of WT or AR strains in minced beef, while in potato slices, D-values in almost all cases were significantly higher in samples which had received heat-shock treatment. In minced beef, the use of a non-selective/overlay recovery medium did not result in higher D-values for any strains, while in potato, significantly higher (P < 0.05) D-values were obtained in most cases. CONCLUSION: The presence or absence of antibiotic resistance genes did not modulate the heat resistance of the strains examined in this study. SIGNIFICANCE AND IMPACT OF THE STUDY: The study demonstrated that heat shock, and the type of media used to determine bacterial numbers during heat processing, can significantly affect the D-values obtained.     

Enhancement of Parainfluenza 2 Virus Hemagglutinin

A comparison was made of the three previously described methods of enhancement of parainfluenza 2 virus hemagglutinin (HA): ultrasonic oscillation, adsorption to erythrocytes and elution (AE), and Tween-ether (TE) treatment. TE treatment resulted in the maximal elevation of HA titer, complete loss of infectivity, and the maximal loss of heat stability of HA. Ultrasonic oscillation caused the least elevation of HA titer, minimal loss of infectivity, and the least loss of heat stability of both HA and infectivity. The effect of AE remained intermediate. It is suggested that the changes produced by these three methods are varying degrees of disruption of virus particles. Growth studies on parainfluenza 2 virus are also presented.     

A Protocol to Assess Insect Resistance to Heat Waves,Applied to Bumblebees (Bombus Latreille, 1802)

Insect decline results from numerous interacting factors including climate change. One of the major phenomena related to climate change is the increase of the frequency of extreme events such as heat waves. Since heat waves are suspected to dramatically increase insect mortality, there is an urgent need to assess their potential impact. Here, we determined and compared the resistance to heat waves of insects under hyperthermic stress through their time before heat stupor (THS) when they are exposed to an extreme temperature (40°C). For this, we used a new experimental standardised device available in the field or in locations close to the field collecting sites. We applied this approach on different Arctic, Boreo-Alpine and Widespread bumblebee species in order to predict consequences of heat waves. Our results show a heat resistance gradient: the heat stress resistance of species with a centred arctic distribution is weaker than the heat resistance of the Boreo-Alpine species with a larger distribution which is itself lower than the heat stress resistance of the ubiquitous species.     

Impacts of a simulated heat wave on composition of a marine community

Extreme events, such as heat waves, are predicted to increase in frequency, duration, and severity as a consequence of climate change. However, global change research generally focuses on increases in mean temperatures and fails to address the potential impacts of increasingly severe heat waves. In addition, climate change may interact with another primary threat to biodiversity, non‐native species invasions. We assessed the impacts of a short‐term heat wave on the marine epibenthic fouling community of Bodega Harbor, California, USA, by exposing experimental mesocosms to a simulated heat wave in the laboratory and then monitoring community development in the field. We hypothesized that (1) juveniles would be more susceptible to heat waves than adults, (2) native species would be more susceptible than non‐native species, and (3) non‐native species would recover more quickly than native species. We observed no effect of the heat wave on juvenile species richness, either initially or during the recovery period, relative to communities at ambient seawater temperatures. In contrast, total adult species richness initially declined in response to the heat wave. Adult community composition also changed in heat‐wave treatments, with non‐natives representing the majority of species and occupying more cover than native species. The reduction in native richness associated with the heat wave persisted through the recovery period, whereas invasive richness was actually higher on heat‐wave versus ambient plates at 95 days. Heat waves have the potential to alter the composition of this community because of species‐, taxon‐, and/or origin‐specific responses; for example, non‐native bryozoans displayed greater resistance than native and non‐native tunicates. Recovery from the heat wave occurred via growth of resistant individuals and larval recruitment. Our study highlights the importance of considering species’ and community responses to heat waves, and not just mean predicted temperature increases, to evaluate the consequences of climate change.     

Thermal inactivation of foot-and-mouth disease viruses in suspension

The heat resistance of foot-and-mouth disease virus (FMDV) strains isolated from outbreaks in Thailand was investigated in phosphate-buffered saline (PBS) at 50, 60, 70, 80, 90, and 100 degrees C. The first-order kinetic model fitted most of the observed linear inactivation curves. The ranges of decimal-reduction time (D value) of FMDV strains at 50, 60, 70, 80, 90, and 100 degrees C were 732 to 1,275 s, 16.37 to 42.00 s, 6.06 to 10.87 s, 2.84 to 5.99 s, 1.65 to 3.18 s, and 1.90 to 2.94 s, respectively. The heat resistances of FMDV strains at lower temperature (50 degrees C) were not serotype specific. The effective inactivating temperature is approximately 60 degrees C. Heat resistances of FMDV strains at 90 and 100 degrees C were not statistically different (P > 0.05), while the FMDV serotype O (OPN) appeared to be the most heat resistant at 60 to 80 degrees C. The other observed inactivation curves were linear with shoulder or tailing (biphasic curves). The shoulder effect was mostly observed at 90 and 100 degrees C, while the tailing effect was mostly observed at 50 to 80 degrees C. The adjusted D values in the case of shoulder and tailing effects did not affect the overall estimated heat resistance of these FMDV strains, so even unadjusted D values of deviant inactivation curves were legitimate. The z values of FMDV serotypes O, A, and Asia 1 were 21.78 to 23.26, 20.75 to 22.79, and 19.87 degrees C, respectively. The z values of FMDV strains studied were not statistically significantly different (P > 0.05). The results of this study indicated that the heat resistance in PBS of FMDV strains from Thailand was much less than had been reported for foreign epidemic FMDV strains.     

Heat resistance of coliform species isolated from cooked ham, snail flesh, and 'bouchées à la reine'

AIMS: In this study, the heat resistance of coliform species isolated from cooked ham and ready-made meals was determined. METHODS AND RESULTS: Thirteen coliform strains belonging to 12 different species were studied using laboratory medium in order to determine delta (first decimal reduction time) and z(T) values (temperature increase leading to a 10-fold reduction of delta) using the Weibull model. For seven strains, delta-values were determined at temperatures ranging from 55 to 60 degrees C, with, delta values between 0.52 and 2.98 min, at 59 degrees C. For the other six strains, lower temperature values were determined with delta-values ranging from 0.47 to 1.64 min at 54 degrees C. z(T) values calculated for the 13 strains were 3.1 to 7.5 degrees C. For eight strains, plotting of the log of survivors was not linear but rather showed shoulders or shoulders and tails. CONCLUSIONS: Coliform species were sensitive to heat treatment with a decimal reduction time under 2 min at 60 degrees C. SIGNIFICANCE AND IMPACT OF THE STUDY: The better knowledge of coliform heat resistance by determining thermal resistance parameters with confidence intervals will be useful for evaluating the efficiency of industrial thermal processes.     

Rheological and flow properties of blood investigated by ultrasound

Ultrasonic waves of 1-15 MHz frequencies easily propagate through soft biological tissues, thus providing qualitative and quantitative information on mechanical and flow properties of blood and red blood cell (RBC) suspensions. Two types of techniques allow to investigate blood behaviors: echographic devices via amplitude detection and Doppler effect based devices via frequency detection of the ultrasonic signal. When ever B mode serves to construct images of tissue slabs from the ultrasonic backscattering coefficient and can give qualitative information on the mechanical properties of blood, A-mode allows to quantify the ultrasonic backscattering coefficient. Ultrasonic Doppler modes also provide both qualitative and quantitative information on blood flow velocity: continuous and pulsed Doppler modes provide curves of blood flow versus time when color Doppler and power Doppler imaging visualize blood flowing in human vessels. Association of echographic and Doppler modes to investigate simultaneously structure and velocity of blood is commercially available. Some examples of results given by such ultrasonic techniques that contribute to characterize, both in vitro and in vivo, structure and flow properties of blood or red blood cell (RBC) suspensions are presented.     

Effects of simulated heat waves on an experimental plant–herbivore–predator food chain

Greater climatic variability and extreme climatic events are currently emerging as two of the most important facets of climate change. Predicting the effects of extreme climatic events, such as heat waves, is a major challenge because they may affect both organisms and trophic interactions, leading to complex responses at the community level. In this study, we set up a simple three‐level food chain composed of a sweet pepper plant, Capsicum annuum; an aphid, Myzus persicae; and a ladybeetle, Coleomegilla maculata, to explore the consequences of simulated heat waves on organism performance, trophic interactions, and population dynamics. We found that (1) heat waves do not affect plant biomass, significantly reduce the abundance and fecundity of aphids, and slightly affect ladybeetle developmental time and biomass, (2) heat waves decrease the impact of ladybeetles on aphid populations but do not modify the effect of aphids on plant biomass, and (3) food chains including predatory ladybeetles are more resistant to heat waves than a simple plant–aphid association, with aphid abundance being less influenced by heat waves in the presence of C. maculata. Our results suggest that more biodiverse ecosystems with predators exerting a strong biotic control are likely to be less influenced by abiotic factors and then more resistant to extreme climatic events than impoverished ecosystems lacking predators. Our study emphasizes the importance of assessing the effects of climatic change on each trophic level as well as on trophic interactions to further our understanding of the stability, resilience, and resistance of ecological communities under climatic forcing.     

控水与补水条件下连续热浪对闽楠光合特性和生长速率的影响

全球变暖背景下,亚热带地区极端气候热浪事件发生频率持续增加。高频热浪及其伴随的高温和干旱复合胁迫将严重影响植物的光合特性,抑制植物的生长,甚至造成死亡。然而,目前亚热带树木光合特性及生长对高频热浪及复合胁迫的响应仍不明确。以亚热带阔叶树种闽楠苗木为研究对象进行了热浪模拟实验,关注了补水和控水处理不同水分环境下连续热浪对闽楠光合特性及生长速率的影响。结果表明,补水处理下闽楠净光合速率(Pn)在单次热浪影响下显著下降了34%,同时水分利用效率显著(WUE)下降,但蒸腾速率(Tr)、气孔导度(Gs)和叶片水汽压亏缺(Leaf_vpd)显著上升(P<0.05)。表明水分充沛的高温环境中闽楠可通过增加蒸腾耗水加速水分蒸散来调节叶片的温度,增强植株光合特性对热浪的抗性。而控水处理下单次热浪处理组Tr和Gs未显著上升,以及Tr与Leaf_vpd在干湿环境下线性拟合的不同斜率,说明水分胁迫会降低叶片降温的效率,加剧热浪对闽楠光合特性的影响。水分和高温的复合胁迫还延长了闽楠光合特性在热浪后的恢复过程,高温胁迫下闽楠Pn在15 d后恢复至未干扰水平,但复合胁迫...     

The effect of combined heat and ultrasound on multicellular tumour spheroids

The effect of combined ultrasound and heat treatments on Chinese hamster multicellular spheroids of varying size was investigated using growth rate, single cell survival and ultrastructural damage as endpoints. Ultrasonic irradiation at 37 degrees C had no effect on the growth rate of 200-730 microns spheroids. Similarly there was no effect on the growth rate of 350 microns spheroids when irradiated during a 60 min exposure to 41.5 degrees C. However, spheroids of 200-700 mm diameter showed growth delay when held at 43 degrees C for 1 h. The effect was enhanced with concomitant ultrasound irradiation but was not dependent on spheroid size. When 200 and 400 microns spheroids held at 43 degrees C for 60 min were irradiated with different ultrasonic intensities a dose-dependent decrease in surviving fraction and a dose-dependent increase in growth delay was obtained. When surviving fraction was plotted as a function of growth delay a good correlation was obtained, suggesting that the combination of heat and ultrasound irradiation does not produce cytostasis in the surviving cells of either 200 or 400 microns spheroids. At the ultrastructural level increased cytoplasmic vacuolation was the only result of ultrasonic irradiation at 37 degrees C. Exposure to 43 degrees C for 60 min was required to elicit thermal damage. This took the form of membrane evagination at the spheroid surface, vacuolation of the cytoplasm, grouping of organelles around the periphery of the nucleus, and fragmentation of the nucleolus. These effects were enhanced with concomitant ultrasonic irradiation but other features were also noted, viz. disaggregation of polyribosomes, dilation of the rough endoplasmic reticulum and blebbing of the nuclear membrane. Damage was independent of spheroid size. These results are in agreement with previous data obtained from single-cell studies. Indicating that there is a non-thermal, non-cavitational component to the cell killing in multicellular spheroids resulting from combined heat and ultrasound treatment.     

Ultrasound induced strain cytoskeleton rearrangement: An experimental and simulation study

Cytoskeleton and specially actin filaments are responsible for mechanical modulation of cellular behavior. These structures could be fluidized in response to transient mechanical cues. Ultrasound devices have been widely used in medicine which their generated ultrasonic waves could disrupt/fluidize actin filaments in cytoskeleton and thus could affect cellular organization. Present research aims at revealing the mechanism of fluidization caused by ultrasound induced strains. First, a numerical simulation was performed to reveal the effect of oscillating ultrasonic pressure on induced deformation in the cell with respect to different cell geometries and exposure conditions. The model revealed that higher pressure and frequencies induce higher levels of strain in the cell. The results also showed that spread cells are more exposed to cytomechanical remodeling due to higher level of ultrasound induced deformations but also the effect of harmonic excitation decreases with spreading. Furthermore, strain values found to be less in the nucleus comparing the value in the cytoplasm, but still these strains can affect the behavior of the cell through mechanotransduction mechanisms. Then, different experimental ultrasound protocols were used to evaluate their effects on cell viability and actin cytoskeleton distribution. Results of Live/Dead assay indicated that high pressure and duration of the exposure had negative effects on the viability of C2C12 cells, while the viability ratio still remained above 85%. In addition, actin fluorescent staining showed that high levels of filament disruption could occur with increasing the pressure. The results of this study shed light on cellular response to mechanical stimuli applied by ultrasonic waves.     

Effect of ultrasonic treatment on the biochemphysical properties of chitosan

Four chitosans with different molecular weights and degrees of deacetylation degree and 28 chitosans derived from these initial chitosans by ultrasonic degradation have been characterized by gel permeation chromatography (GPC), FT-IR spectroscopy, X-ray diffraction and titrimetric analyses. Antimicrobial activities were investigated against E. coli and S. aureus using an inhibitory rate technique. The results showed that ultrasonic treatment decreased the molecular weight of chitosan, and that chitosan with higher molecular weight and higher DD was more easily degraded. The polydispersity decreased with ultrasonic treatment time, which was in linear relationship with the decrease of molecular weight. Ultrasonic degradation changed the DD of initial chitosan with a lower DD (<90%), but not the DD of the initials chitosan with a higher DD (>90%). The increased crystallinity of ultrasonically treated chitosan indicated that ultrasonic treatment changed the physical structure of chitosan, mainly due to the decrease of molecular weight. Ultrasonic treatment enhanced the antimicrobial activity of chitosan, mainly due to the decrease of molecular weight.     

Enhancement of Ultrasonic Seed Treatment on Yield,Grain Quality Characters,and 2-Acetyl-1-Pyrroline Biosynthesis in Different Fragrant Rice Genotypes

Fragrant rice is popular for the good grain quality and special aroma. The present study conducted a field experiment to investigate the effects of ultrasonic seed treatment on grain yield, quality characters, physiological properties and aroma biosynthesis of different fragrant rice genotypes. The seeds of three fragrant rice genotypes were exposed to 1 min of ultrasonic vibration and then cultivated in paddy field. The results of present study showed that ultrasonic seed treatment increased grain yield of all fragrant rice genotypes but the responses of yield formation to ultrasonic were varied with different genotypes. Compared with control, ultrasonic seed treatment increased grain 2-acetyl-1-pyrroline (2-AP, the key component of fragrant rice aroma) content by 13.40%–44.88%. Ultrasonic seed treatment also reduced the crude protein contents in grains. The head rice rate, rice length, chalky rice rate, and chalkiness degree were influenced by ultrasonic for one or two fragrant rice genotypes. The activities of peroxidase and superoxide dismutase were also enhanced due to ultrasonic seed treatment. In conclusion, ultrasonic seed treatment increased grain, regulated grain aroma and quality, and improved stress resistance of fragrant rice varieties.     

The Incidence of Organisms of the Genus Clostridium in Vacuum Packed Fish in the United Kingdom

Of a total of 224 samples of commercial vacuum packed fish products examined for the presence of Clostridium spp., 151 gave positive cultures. The predominant species was Cl. welchii; Cl. botulinum was not isolated. Spore suspensions of 29 strains of Cl. welchii were tested for heat resistance; none survived treatment for 5 min at 100°.