Effects of Thermotherapy Treatments on Systemic Agrobacterium vitis in Dormant Grape Cuttings

Agrobacterium tumefaciens biovar 3 (A. vitis) was eradicated or reduced below the level of detection in dormant cuttings of grape [Vitis vinifera cultivar (cv.) Thompson seedless] and rootstock NAZ3 (V. vinifera × V. rupestris) by hot‐water treatment (exposure to 50°C for 30 min). Strains of A. vitis varied in their sensitivity to heat, but were generally more sensitive than strains of A. tumefaciens biovar 1 or biovar 2. Populations of about 10³ CFU/ml in broth were killed by a 30‐min treatment. Biovar 1 strains were apparently unaffected by 50°C, even when exposed for 30 min. Non‐tumorigenic biovar 1 strains were recovered from hot‐water‐treated cuttings. An assessment was also made of the effect of treatment on growth parameters of cuttings in a field nursery after 9 months. The effect of hot‐water treatment on the vitality and growth of vines varied with different cultivars or rootstocks. The number and length of canes, root dry weight and length and diameter of trunks, increased in most instances. Treatment and time usually did affect bud survival and, in most cases, increased the level of callus formation at the base of the cuttings. Hot‐water treatment may offer a simple, effective, economical and environmentally safe means of eradicating A. vitis from dormant grape cuttings.     

Combining heat treatment and subsequent incubation temperature to prevent growth from spores of non-proteolytic Clostridium botulinum

Refrigerated processed foods of extended durability rely on a mild heat treatment combined with refrigerated storage to ensure microbiological safety and quality. The principal microbiological safety risk in foods of this type is non-proteolytic Clostridium botulinum. In this article the combined effect of mild heat treatment and refrigerated storage on the time to growth and probability of growth from spores of non-proteolytic Cl. botulinum is described. Spores of non-proteolytic Cl. botulinum (two strains each of type B, E and F) were heated at 90°C for between 0 and 60 min and subsequently incubated at 5°, 10° or 30°C in PYGS broth in the presence or absence of lysozyme. The number of spores that resulted in turbidity depended on the combination of heat treatment, incubation time and incubation temperature they received. Heating at 90°C for 1 or more min ensured a 10⁶ reduction when spores were subsequently incubated at 5°C for up to 23 weeks. Heating at 90°C for 60 min ensured a 10⁶ reduction over 23 weeks when subsequent incubation was at 10°C in the presence of added lysozyme. The same treatment did not reduce the spore population by 10⁶ when subsequent incubation was at 30°C.     

Effect of air and soil temperature on water balance of jojoba growing under controlled conditions

Jojoba [ Simmondsia chinensis (Link) Schneider] cuttings were grown in pots under constant light intensity and vapour pressure deficit at wir temperatures of 18 and 27°C in climate-controlled cabinets. Leaf conductance and transpiration rate decreased exponentially as the xylen water potential (Ψ_x) decreased concurrently with the drying out of the soil. At high Ψ_x'leaf conductance and transpiration rate were much higher at the higher air temperature, and as Ψ_x declined both parameters decreased more rapidly at 27°C than at 18°C. When soil temperatures were decreased from 27 to 13°C, leaf water potential was not affected at either air temperatures, but transpiration rate was reduced. A linear negative correlation was found between transpiration rates and soil temperatures. It is suggested that the low soil temperature may restrict reducion of water flux in turn reduces stomatal conductance and transpiration without affecting the water potential in the shoot. The releavance of the response to changes in soil or air temperature to the performance of the plant in its semi-arid habitat is discussed.     

Dynamics of extractable carbohydrates in Pisum sativum. III. The effect of IAA and temperature on content and translocation of carbohydrates in pea cuttings during rooting

The number of roots formed in cuttings of pea ( Pisum sativum L. cv. Alaska) was regulated both by the temperature and by IAA, whereas the time to the appearance of the first roots was regulated only by the temperature. Cuttings treated with 10^-3 M IAA had a smaller content of extractable carbohydrates than the control ones irrespective of the temperature. In the bases of cuttings rooted at 25°C the content of extractable carbohydrates was lower than in those rooted at 15°C. Cuttings treated with IAA showed up to elevenfold increase of extractable carbohydrates in the bases at day 3. This increase of soluble sugars was not correlated with the number of roots formed or the speed of rooting. It is concluded that IAA affects the accumulation of carbohydrates, and this is not connected with the rooting ability of the cuttings.     

Morning glory systemically accumulates scopoletin and scopolin after interaction with Fusarium oxysporum

An isolate of non-pathogenic Fusarium, Fusarium oxysporum 101-2 (NPF), induces resistance in the cuttings of morning glory against Fusarium wilt caused by F. oxysporum f. sp. batatas O-17 (PF). The effect of NPF on phenylpropanoid metabolism in morning glory cuttings was studied. It was found that morning glory tissues responded to treatment with NPF bud-cell suspension (108 bud-cells/ml) with the activation of phenylalanine ammonia-lyase (PAL). PAL activity was induced faster and greater in the NPF-treated cuttings compared to cuttings of a distilled water control. High performance liquid chromatography analysis of the extract from tissues of morning glory cuttings after NPF treatment showed a quicker induction of scopoletin and scopolin synthesis than that seen in the control cuttings. PF also the induced synthesis of these compounds at 10(5) bud-cells/ml, but inhibited it at 10(8) bud-cells/ml. Possibly PF produced constituent(s) that elicited the inhibitory effect on induction of the resistance reaction. These compounds could potentially be useful as markers to detect early beginning interactions between Fusarium and morning glory tissues cuttings.     

Heat treatment to decrease chilling injury in tomato fruit. Effects on lipids, pericarp lesions and fungal growth

Tomato fruits are sensitive to low temperature and develop chilling injury, while at nonchilling temperatures they ripen rapidly. Previously, a hot-air treatment was found to reduce the sensitivity of the fruit to low temperatures. In the present study hot air was compared to hot water and their effects on reducing chilling injury and fungal decay were investigated. Tomatoes ( Lycopersicon esculentum cv. Daniella) at the breaker stage were subjected to hot air, 48 h at 38°C, or various hot water dips, 30 min at 40°C or 2 min at 46, 48 or 50°C, before holding at 2°C. The unheated tomatoes developed chilling injury and fungal infections at 2°C, but not at 12°C. All the heat treatments reduced chilling injury and decay in tomatoes held for 3 weeks at 2°C. The outer pericarp tissue of heated tomatoes had higher phospholipid and lower sterol contents than unheated tomatoes. Heated tomatoes also had less saturated fatty acids than unheated tomatoes held at 2°C, but not at 12°C. Scanning electron micrograph observations showed that all the fruits had microcracks in their surface, but the unheated chilled tomatoes had also fungal growth in the cracks, while those of the heated tomato fruit did not. In the areas of chilling injury collapsed cells were present under the peel and could also support pathogen development. It is suggested that the heat treatment institutes a response to high temperature stress in the fruit tissue that leads to strengthened membranes. This prevents the loss of function and cell collapse which was found in the chilling-injured areas of affected fruit.     

Rooting of etiolated stem segments of Populus robusta – interaction of temperature, catechol and sucrose in the presence of IAA

Etiolated stem segments of Populus robusta Schneid. were cultured in test solutions each containing 2.0 mg/1 of IAA in combination with varying concentrations of catechol or sucrose or both at 10 ± 2°C, 25 ± 1°C and 35 ± 1°C in the dark. Cuttings did not root in catechol in the presence of IAA alone at 10 ± 2°C. There was no mortality in this culture solution at this temperature. However, cuttings rooted in this culture solution at 25 ± 1°C and 35 ± 1°C and also showed mortality, which was more severe at 35 ± 1°C than at 25 ± 1°C and more severe at higher than at lower concentrations of catechol. Rooting occurred on cuttings in catechol in the presence of IAA and sucrose even at 10 ± 2°C and was strongly promoted at 25 ± 1°C and 35 ± 1°C. The mortality of segments caused by catechol was markedly lowered in the presence of sucrose at these temperatures. Sucrose thus antagonised the toxic effect of catechol.     

Heat resistance of Cronobacter species (Enterobacter sakazakii) in milk and special feeding formula

Aim:  To determine D - and z -values of Cronobacter species ( Enterobacter sakazakii ) in different reconstituted milk and special feeding formula and the effect of reconstitution of powdered milk and special feeding formula with hot water on the survival of the micro-organism.
Methods and Results:  Five Cronobacter species (four C. sakazakii isolates and C. muytjensii ) were heated in reconstituted milk or feeding formula pre-equilibrated at 52–58°C for various times or mixed with powdered milk or feeding formula prior to reconstitution with water at 60–100°C. The D -values of Cronobacter at 52–58°C were significantly higher in whole milk (22·10–0·68 min) than in low fat (15·87–0·62 min) or skim milk (15·30–0·51 min) and significantly higher in lactose-free formula (19·57–0·66 min) than in soy protein formula (17·22–0·63 min). The z -values of Cronobacter in reconstituted milk or feeding formula ranged from 4·01°C to 4·39°C. Water heated to ≥70°C and added to powdered milk and formula resulted in a > 4 log₁₀ reduction of Cronobacter .
Conclusions:  The heat resistance of Cronobacter should not allow the survival of the pathogen during normal pasteurization treatment. The use of hot water (≥70°C) during reconstitution appears to be an effective means to reduce the risk of Cronobacter in these products.
Significance and Impact of the Study:  This study supports existing data available to regulatory agencies and milk producers that recommended heat treatments are sufficient to substantially reduce risk from Cronobacter which may be present in these products.     

Induction of triploidy in offspring of gilthead seabream (Sparus aurata) by means of heat shock

By using heat shock, we have induced triploidy in the offspring of gilthead seabream ( Sparus aurata ). We have verified that the effectiveness of the treatment in this marine species depends not only on the three factors habitually considered (incubation time, treatment intensity and treatment time), but also on the salinity of the treatment water. The most effective treatment to provoke triploidy in this species is to apply a heat shock of 34 °C for 10 min without an incubation period and with water salinity at 30:1000.     

THE PREVENTION OF MICROBIAL SPOILAGE IN WHOLE SHELL EGGS BY HEAT TREATMENT METHODS

SUMMARY: The prevention of microbial spoilage in whole shell eggs by heat treatment methods has been investigated. After shell eggs had been immersed for 3 min in water at 144·5 ± 0·5°F the incidence of microbial rotting was reduced by over 90%. Consistent reductions in spoilage losses were obtained with farm steeped eggs, eggs produced under commercial conditions, and eggs held in cold storage for 6 months. Heat treatment efficiently controlled green, sour and black rots in stored eggs, but although reductions were noted in the number of mould rots, it cannot be guaranteed that this type of spoilage can be controlled with similar certainty.
Internal quality in heat treated eggs was satisfactory. During protracted holding in cold storage, heat treatment appeared to result in a better internal egg quality and a more stable keeping quality, than when untreated eggs were held under similar conditions.     

Causes of Centrifugal Root Promotion

Role of water in centrifugal root promotion, effect of centrifugation on ethylene concentration of cuttings, and effect of ethylene on root formation were investigated using Salix jragilis softwood cuttings. When cuttings were centrifuged with water, more roots formed with increasing water depth. Soaking of cuttings upright in water for 24 h stimulated root formation and produced more roots as the depth of water was increased, with submerged cuttings producing the highest number of roots. Soaking of cuttings upright in hot water for 1 h also stimulated root formation with the best root formation occurring at 40°C. Submerging in, or centrifuging with water increased ethylene concentration in cuttings. Ethylene gas and Ethrel treatments stimulated root formation. It is suggested that submerging in water increases the ethylene concentration in Salix fragilis softwood cuttings which in turn stimulates root formation of the cuttings. Centrifuging cuttings with water increased water content of the cuttings. It is suggested that this increase in water content plays a role similar to the submersion of cuttings as described above.     

Effect of bacterial suspensions on vascular occlusion in stems of cut rose flowers

A suspension of Pseudomans aeruginosa at 5 × 10⁹ cfu/ml was either left untreated, pasteurized (15 min, 70°C) or autoclaved (15 min, 121°C). Stems of cut rose flowers ( Rosa hybrida L. cv. Sonia) placed in these solutions for 0·5–4·0 h showed decreased water uptake and a reduction of hydraulic conductance of the basal 5·0 cm stem segment. No difference was found between the treatments. When the pasteurized or autoclaved solutions were left at 4°C for 7 days, bacterial cells had autolyzed, and stems placed in these suspensions showed lower water uptake and hydraulic conductance than stems in freshly prepared solutions. The results show that living and non-living bacteria have the same effect on vascular occlusion, and indicate that hydraulic conductance was more reduced when the average size of the particles was smaller than that of bacterial cells. Vascular blockage occurred within 30 min after the start of treatment, apparently due to a physical effect rather than a physiological response of the stem tissue.     

Effects of combinational treatment with ethephon and indole-3-butyric acid on adventitious rooting of <Emphasis Type="Italic">Pinus thunbergii</Emphasis> cuttings

Adventitious rooting is essential for cutting propagation of pine wilt-resistant Pinus thunbergii. To examine a variety of adventitious rooting potentials among donor plants, cuttings were taken from 31 seedlings within a half-sib family. Rooting abilities of cuttings from each seedling ranged from 0 to 100%. When 11 ortets and 11 ramets (clonally propagated from each ortet) were used as donor plants, there was a positive correlation between rooting abilities of cuttings from ortets and ramets, suggesting that adventitious rooting is dependent on genetic factors in the donor plants. To promote adventitious rooting of cuttings by hormonal treatment, we examined the effect of soaking time in Oxyberon (19.7 mM indole-3-butyric acid (IBA) solution) on rooting. Ten minutes was the best soaking time for rooted cuttings to produce more adventitious roots without impairing normal growth. When cuttings were soaked in Ethrel diluent (69.2 μM ethephon) for 24 h before soaking in Oxyberon for 10 min, a significantly higher rooting ability was observed than those soaked in Oxyberon alone. Ethrel on its own barely affected rooting ability. The positive effect of the combinational treatment was confirmed in a similar experiment using authentic ethephon and IBA instead of Ethrel and Oxyberon. When cuttings were soaked in a mixture of ethephon and silver thiosulfate (STS), an ethylene action inhibitor, before IBA-soaking, the effect was partially diminished compared with combinational treatment without STS. These findings suggest that ethylene action caused by ethephon treatment promotes IBA-mediated adventitious rooting of P. thunbergii cuttings.     

Preliminary Microbiological Studies on the Preservation of Palm Wine

Palm wine is essentially a heavy suspension of yeasts and bacteria in fermenting palm sap. The water extract of a local preservative, from the bark of Sacoglottis gabonensis , failed to inhibit several yeasts and bacteria from palm wine. Sodium metabisulphite, diethylpyrocarbonate (DEPC) and sorbic acid inhibited to varying extents micro-organisms in palm wine. Sorbic acid was thought to be the most suitable preservative tested since DEPC left a residual pungent smell and sodium metabisulphite was not suitable because it is unacceptable to man above 0.35 mg/kg of body weight (Mossel, 1971). Pasteurization at 70° for 30 min was more effective than any of the chemical methods in reducing the microbial load of the wine. It is suggested that pasteurization at 70° for 30 min combined with subsequent treatment with sorbic acid may prove useful as a means of preserving palm wine.     

The histopathology of salmon tagging

Hatchery reared Atlantic salmon ( Salmo salar L.) parr were tagged with the standard I.C.E.S./I.C.N.A.F. salmon tag at water temperatures of 15°C and 6°C. They were then subjected to swimming trials in a tunnel respirometer. Following this treatment the 15°C fish developed necrotic infarcts around the tag insertions. Provided they did not become infected these lesions healed, but if the fish were retested 9 days later, the wound enlarged, became extremely inflamed, and the fish died soon after. This effect was not seen in 6°C fish.     

Chemical composition and characterization of a galactomannoglucan from Gliocladium viride wall material

The following fractions were obtained from the wall material of Gliocladium viride : F1 (27.5%), a glucan, containing xylose, mannose and galactose, coluble in 1 M NaOH at 20°C; F2 (6.7%), a β-glucan-chitin complex, solubilized with 1 M NaOH at 20°C from the previous residue left overnight at −20°C; F3 (8.1%), a glucan, containing mannose and galactose solubilized with 1 M NaOH at 70°C; and F4, the insoluble residue, a β-glucan-chitin complex similar to F2, amounting to 31.3% of the wall material.
F1 was extracted with distilled water. The soluble material (F1S) was a galactomannoglucan (54.7%) and the inscluble (F1P) a glucan (45.3%). Periodate oxidation revealed the presence of glycerol, erythritol, threitol, ribitol, arabitol, mannose, galactose and glucose in F1S, and glycerol and glucose as the main components in F1P. The fractions obtained when F1S was purified through Sepharose CL6B, were methylated.     

Sensitivity of Petri disease pathogens to hot-water treatments in vitro

Petri disease pathogens ( Phaeoacremonium spp. and Phaeomoniella chlamydospora ) are able to colonize the vessels in the xylem of grapevine propagating material. Hot-water treatment (HWT) protocols at 50°C for 30 min have been applied in grapevine nurseries to control these pathogens with variable results. The effect of HWT in vitro at higher temperatures on Pa.   chlamydospora , Phaeoacremonium aleophilum and Phaeoacremonium parasiticum isolates was determined by placing conidial suspensions and plugs of agar with mycelia in Eppendorf vials and incubated in hot water at 49, 50, 51, 52, 53 or 54°C for 30, 45 or 60 min. Conidial germination and the colony growth rate decreased with increased temperature and time combinations. Pa.   chlamydospora was more sensitive than Phaeoacremonium spp. to the experimental temperatures for all periods of time. Pa.   chlamydospora tolerated temperatures of 53°C, while Pm.   aleophilum and Pm.   parasiticum tolerated temperatures of 54°C, although the treatments above 51–52°C drastically reduced conidial germination and mycelial growth. These results highlight the need to develop HWT using temperatures above 51°C to reduce the incidence of fungal infections and ensure high-quality propagating material for grapevine growers.     

Dynamics of extractable carbohydrates in Pisum sativum. II. Carbohydrate content and photosynthesis of pea cuttings in relation to irradiance and stock plant temperature and genotype

Rooting ability was studied for cuttings derived from stock plants of wild type pea seedlings and seedings of two mutants deficient in photosystem II activity and chlorophyll. Stock plants were grown at 15, 20, 25 or 30°C at 38 W m^-2. Cuttings were rooted at 20°C and at an irradiance of 16 or 38 W m^-2. The rooting ability seemed to be correlated with the initial carbohydrate content only at 38 W m^-2. Based on the findings of the present study it may be concluded that for pea seedlings the growth temperature is more important than photosynthesis as regards accumulation of extractable carbohydrates. During the rooting period carbohydrates are necessary for root formation, but the effect of the iradiance on the number of roots formed is not mediated by the carbohydrate content. Under specific rooting conditions it is possible to correlate the initial carbohydrate content with the rooting capacity of the cuttings within a phenotype, but not always when different phenotypes are considered. The results indicate a connection between the metabolic activity of the cuttings and their ability to form adventitious roots.     

A Model System for Testing the Microbiological Stability of Foods Processed in Laminated Flexible Pouches

A suitable time temperature process for packaging small potatoes in flexible pouches is described. Spores of Bacillus stearothermophilus were inoculated onto the surface of peeled potatoes in flexible pouches. These were evacuated, heat sealed and heated in a steam retort modified to allow a water cooking process with an air overpressure of ca. 68·95 kPa (10 lbf/in²). The D and z values determined in the retort were confirmed by parallel heat-resistance tests with spores in glass ampoules held in an oil bath. Heat treatment at 121·1 °C for 20 min in the retort satisfactorily killed test inocula of spores without overcooking the potatoes. Uninoculated peeled potatoes with a natural level of contamination ( ca. 4 x 10³ spores/sealed pouch) were treated in the retort for different times at 121·1 °C and then incubated at 30 °C for 6 months. No pouches heated for 17–25 min showed microbial growth. This procedure may be applied to any type of food if the numbers of naturally occurring heat-resistant contaminants can be related to the number and heat resistance of a suitable test micro-organism.     

Cadmium and copper uptake and distribution in Mediterranean tree seedlings

Provenances of seedlings of Scots pine ( Pinus sylvestris L.) from northern Finland (65°N) and cuttings from trees of the same species about 30 to 40 years in age were studied in 3 artificial deacclimation experiments during the winter and spring of 1994. The deacclimation conditions varied in terms of temperature rise and photoperiods. Frost resistance and the pH of the cell effusate were studied in order to investigate the possibility of a connection between them. Complete deacclimation was achieved, as shown by a frost resistance of –60°C to –65°C in the seedlings and –8O°C in the cuttings at the beginning of the experiment and decreased to above –10°C by the end. The frost resistance and pH of the cell effusate responded significantly to the increase in temperature that marked the onset of the deacclimation process, whereas the response to different photoperiods was only slight. A highly significant correlation was found between frost resistance and the pH of the cell effusate. These variables were also significantly correlated with the freezing test temperature, indicating the condition of plasmalemma and tonoplast; and with sampling day, indicating the response to deacclimation. In the light of these findings, it is concluded that the pH of the cell effusate with certain reservations, can be used as a time saving, alternative viability test when determining frost resistance.