Effect of different cold storage periods on parasitization performance of Trichogramma cacoeciae (Hymenoptera,Trichogrammatidae) on eggs of Ephestia kuehniella (Lepidoptera,Pyralidae)

The parasitism rates by Trichogramma cacoeciae Marchal (Hymenoptera, Trichogrammatidae) using Ephestia kuehniella Zell. (Lepidoptera, Pyralidae) eggs held at 0, 4 and 8°C and for up to 31 days was measured. Parasitism was lowest on eggs held at 8°C and highest on eggs held at 0°C. The highest parasitism, 97.8%, was measured for parasitoids attacking eggs held for 3 days and stored at 0°C. Parasitism of eggs stored at all three temperatures decreased with increasing duration of storage. The number of T. cacoeciae successfully developing and emerging as adults after storage in E. kuehniella eggs held at 0, 4 and 8°C was measured. Parasitoid emergence was >83% from E. kuehniella eggs stored at 8°C for 3 weeks. Storage at 0°C caused a significant decline in parasitoid emergence after 2 weeks (P<0.05). Storage at 0°C for more than 4 weeks reduced fecundity by 50%. T. cacoeciae parasitized the highest number of E. kuehniella eggs 1 day after adult emergence. The oviposition period lasted 6–7 days, although the parasitoids lived up to 13–14 days. Impact of storage time and temperature on parasitism rates by T. cacoeciae stored while in E. kuehniella eggs was measured. As storage time and temperature increased, subsequent parasitism rates of resulting adult T. cacoeciae decreased. Eggs of E. kuehniella can be stored at 0°C for up to 31 days. Trichogramma cacoeciae developing in eggs of E. kuehniella can be stored at 4°C for up to 5 weeks prior to release.     

Effect of temperature and ripening stages on membrane integrity of fresh-cut tomatoes

The shelf-life of fresh-cut tomatoes mainly depends on loss of tissue integrity and firmness that occurs also in intact fruits after long-term cold storage due to chilling injury. Round-fruit tomatoes (Solanum lycopersicum L.) cv. Jama were stored in 1.1-L plastic (polyethylene) fresh-cut produce containers as 10.0-mm-thick tomato slices and as intact tomatoes at 4 ± 0.5 °C. The aim of this work was to study the loss of membrane integrity and biochemical processes involved in membrane disruption. Electrolyte leakage and lipid peroxidation were studied at different stages of maturity: mature green, pink (PK), fully ripe and two different storage temperatures: 4 and 15 °C. The tomato slices of PK stage stored at 4 °C did not show changes for both parameters, while significant increase in membrane leakage and lipid peroxidation was observed at 15 °C, especially after 24 h of storage. The enzymes showed a simultaneous increase in their activities with a rise in electrolyte leakage and lipid peroxidation after 7 days of storage. Finally, phospholipase C (PLC) and phospholipase D (PLD) were investigated for intact fruit and tomato slices stored at 4 °C. The PLC had higher activity compared with PLD. In conclusion, the loss of membrane integrity in fresh-cut tomatoes is mainly affected by ripening stages, storage temperature and duration. The wounds enhance the PLC and PLD activities and they play a role late during storage.     

In vitro response of encapsulated and non-encapsulated somatic embryos of Kinnow mandarin (Citrus nobilis Lour × C. deliciosa Tenora)

The aim of present investigation was to study the effect of storage conditions on percentage germination of encapsulated and non-encapsulated somatic embryos of Kinnow mandarin (Citrus nobilis Lour × C. deliciosa Tenora). Different batches of encapsulated and non-encapsulated embryos were preserved at room temperature, 4°C, in liquid nitrogen as such and by embedding in liquid paraffin. In the encapsulated somatic embryos stored at room temperature in sealed Petri plates, percentage of germination was 24.99%, but 5.55% in non-encapsulated embryos after 3 days of storage. Encapsulated embryos stored in vials containing liquid paraffin at room temperature were germinated at 18.05% after 60 days of storage, while it was 13.88% in non-encapsulated embryos after 45 days of storage. Encapsulated somatic embryos stored at 4°C in sealed Petri plates remained viable for up to 75 days with 6.94% germination, whereas non-encapsulated embryos remained viable for up to 45 days with 24.99% germination. Encapsulated embryos stored at 4°C in vials filled with paraffin germinated at 11.11% after 120 days of storage, but 5.55% in non-encapsulated embryos after 90 days of storage. Encapsulated and non-encapsulated embryos stored in liquid nitrogen showed 58.33 and 51.38% survival, respectively, after 7 months of storage. The plantlets developed from these embryos were transplanted after acclimatization and are growing normal.     

Viability and virulence of blastospores of Metarhizium anisopliae (Metch.) Sorokin after storage in various liquids at different temperatures

Blastospores of three strains of Metarhizium anisopliae were stored in 18 liquids at 4°C, 20°C and 35°C for 18 weeks, 12 weeks or 9 days respectively. Viability was quantified by determination of their germination. In bioassays the virulence of stored blastospores was studied using adults and third instars of Locusta migratoria migratorioides (R. & F.) and compared to those of freshly produced blastospores and conidia. Generally, there was great variability in the viability of blastospores, depending on the fungal strain and the liquids used. Blastospores survived best at 4°C in 10% hydroxyethyl starch; for example, germination of M. anisopliae strain 97 still amounted to more than 80% after storage for 18 weeks. Other suitable liquids were deionized water, 25% Ringer's solution and 1% sodium alginate. The viability of blastospores stored at 20°C was considerably shorter than at 4°C. During storage for 12 weeks at 20°C the best protective liquids for M. anisopliae strain 97 were 25% Ringer's solution (43% germination), deionized water (23%) and 10% hydroxyethyl starch (23%). At 35°C, 45% of M. anisopliae strain 97 blastospores still germinated after storage for 7 days in 25% glycerol. The bioassays revealed that the virulence of blastospores after storage was comparable to that of fresh ones and even better than that of fresh conidia. In general, the LT₅₀ was about 4–6 days at an alternating day/night temperature of 28/20°C.     

Preservation of Phakopsora pachyrhizi Uredospores

This study compared different temperatures and dormancy‐reversion procedures for preservation of Phakopsora pachyrhizi uredospores. The storage temperatures tested were room temperature, 5°C, ?20°C and ?80°C. Dehydrated and non‐dehydrated uredospores were used, and evaluations for germination (%) and infectivity (no. of lesions/cm²) were made with fresh harvested spores and after 15, 29, 76, 154 and 231 days of storage. The dormancy‐reversion procedures evaluated were thermal shock (40°C/5 min) followed or not by hydration (moist chamber/24 h). Uredospores stored at room temperature were viable only up to a month of storage, regardless of their hydration condition. Survival of uredospores increased with storage at lower temperatures. Dehydration of uredospores prior to storage increased their viability, mainly for uredospores stored at 5°C, ?20°C and ?80°C. At 5°C and ?20°C, dehydrated uredospores showed increases in viability of at least 47 and 127 days, respectively, compared to non‐dehydrated spores. Uredospore germination and infectivity after storage for 231 days (7.7 months), could only be observed at ?80°C, for both hydration conditions. At this storage temperature, dehydrated and non‐dehydrated uredospores exhibited 56 and 28% of germination at the end of the experiment, respectively. Storage at ?80°C also maintained uredospore infectivity, based upon levels of infection frequency, for both hydration conditions. Among the dormancy‐reversion treatments applied to spores stored at ?80°C, those involving hydration allowed recoveries of 85 to 92% of the initial germination.     

Gluconeogenesis in the rat renal cortex after warm ischemia and hypothermic storage

Rat kidney cortex slices were tested for their gluconeogenic capacity after the kidney has been either subjected to warm ischemia or flushed with and stored in cold hyperosmotic media. Kidneys damaged by warm ischemia for up to 60 min did not lose their ability to convert pyruvate to glucose. However, they then rapidly lost this capacity so that after 2 hr of ischemia they were devoid of activity. This observation closely corresponded to survival of partially nephrectomized rats whose remaining kidney had been treated in a similar manner. Cortex slices obtained from kidneys flushed and stored in cold hyperosmotic media were found to lose most of their gluconeogenic capacity after 3 days of storage.     

Cold tolerance of coccons ofAllorhogas pyralophagus [Hym.: Braconidae]

Cold tolerance studies were conducted under laboratory conditions for coccons ofAllorhogas pyralophagus Marsh, a Mexican parasitoid of graminaceous borers. By storage, developmental time (from cocoon to adult emergence) could be extended by 2 to 6 times. However, cocoons stored for more than 14 days at 2°C failed to survive, while at 5 and 10°C, about 50% emergence was recorded for upto 21 days of storage. With respect to survival and adult longevity, 10°C seemed to be the most suitable storage temperature. Pre-emergence period was also significantly increased by storing cocoons for 21 to 35 days at this temperature. Sex-ratio of emerging adults was not significantly affected by storage. Fecundity was adversely affected in all the treatments except in the case of females emerging from cocoons stored at 5°C for 7 days. The progeny of parasitoids which emerged from cocoons stored at 5 and 10°C for 35 days consisted of only males. It is clear from the present study thatA. pyralophagus cocoons are more amenable to short-term storage. Contribution No. 285/87 of the Indian Institute of Horticultural Research, Bangalore-560 089.     

Survival of Conidia of Clonostachys rosea on Stored Barley Seeds and Their Biocontrol Efficacy Against Seed-borne Bipolaris sorokiniana

Survival and biocontrol activity of Clonostachys rosea (isolate IK726) conidia during storage on barley seeds were investigated. The initial density of colony forming conidia on seed was 4 &#50 10 3 to 9 &#50 10 4 colony forming units (cfu)/seed. After 5 months storage at 4&#176;C, the density decreased by less than one order of magnitude and the biocontrol efficacy against seedling blight caused by seed-borne Bipolaris sorokiniana was maintained at a significantly high level ( > 80% disease reduction) for > 5 months. Conidial survival on seeds stored at 20&#176;C declined more rapidly than at 4&#176;C, and biocontrol efficacy was significantly reduced after 3-5 months. However, conidia produced on solid media over 20 days survived better than conidia produced in liquid culture and conidia from solid media produced over 12 days. In contrast, when seeds treated with conidia were packed with silica gel and stored at 20&#176;C, the cfu density decreased by less than one order of magnitude after 5 months and the biocontrol efficacy was still high after 6 months. A dose-response curve revealed that 103 cfu/seed were needed for 80% control of seedling blight. Similar control was obtained in storage experiments when approximately 103 cfu/seed were recovered from seed, indicating that conidia which survived also retained a high ability to control disease.     

Low Temperature Storage of <Emphasis Type="Italic">Telenomus remus</Emphasis> (Nixon) (Hymenoptera: Platygastridae) and its Factitious Host <Emphasis Type="Italic">Corcyra cephalonica</Emphasis> (Stainton) (Lepidoptera: Pyralidae)

We conducted three bioassays to evaluate the effect of low-temperature storage of eggs (host) and pupae and adults (parasitoid) on the biology and parasitism capacity of the egg parasitoid Telenomus remus (Nixon) (Hymenoptera: Platygastridae). Viable stored Corcyra cephalonica (Stainton) (Lepidoptera: Pyralidae) eggs were parasitized to the same degree or even higher than fresh eggs when stored until 14 days at 5°C or until 21 days at 10°C. In contrast, the percentage of parasitized sterilized eggs was equal to the control only when stored for 7 and 14 days. Survival of T. remus pupae declined with storage time at both studied temperatures (5 and 10°C). However, after 7 days of storage, survival of pupae was still 86.3 and 64.9% at 10 and 5°C, respectively. The number of adult male survivors remained similar until the fourth storage day at both 5 and 10°C. In contrast, female survival did not differ until day 8 at 10°C or day 6 at 5°C. Parasitism capacity of stored adults was not altered by storage compared with the control. Therefore, we conclude that the maximal storage time at 10°C is 21 days for viable C. cephalonica eggs and 7 days for T. remus pupae, while parasitoid adults should not be stored for more than 4 days at either 5 or 10°C.     

The composition of fresh and stored oyster mushrooms (Pleurotus ostreatus)

Soluble carbohydrate, protein, polysaccharide and cell wall composition were assayed in freshly harvested Pleurotus ostreatus sporophores and those stored for 4 days at 2° or 18°. Mannitol and trehalose were present at 1.8 and 6.5% dry wt respectively in fresh sporophores, and at reduced levels in those stored at 18°. In sporophores stored at 2°, trehalose levels increased by up to 122%. Soluble polysaccharide appeared to be composed of glycogen-like material, which was susceptible to post-harvest breakdown, and components containing mannose and other sugars. The total protein content was 42% dry wt; no protein degradation was seen in sporophores stored at 2°, but about 25% of the protein disappeared during storage at 18°. Cell wall polysaccharide was utilised during storage. Respiration rate was about 8–10 ml CO₂/g dry wt/hr at harvest and declined to about 5 ml/g dry wt/hr after 40 hr storage at 18°.     

Prior frozen storage enhances the effect of edible coatings against Listeria monocytogenes on cold-smoked salmon during subsequent refrigerated storage

Aims: Listeria monocytogenes is a major safety concern for ready‐to‐eat foods. The overall objective of this study was to investigate whether prior frozen storage could enhance the efficacy of edible coatings against L. monocytogenes on cold‐smoked salmon during subsequent refrigerated storage. Methods and Results: A formulation consisting of sodium lactate (SL, 1·2–2·4%) and sodium diacetate (SD, 0·125–0·25%) or 2·5% Opti.Form (a commercial formulation of SL and SD) was incorporated into each of five edible coatings: alginate, κ‐carrageenan, pectin, gelatin and starch. The coatings were applied onto the surface of cold‐smoked salmon slices inoculated with L. monocytogenes at a level of 500 CFU cm^?2. In the first phase, the slices were first frozen at ?18°C for 6 days and stored at 22°C for 6 days. Alginate, gelatin and starch appeared to be the most effective carriers. In the second phase, cold‐smoked salmon slices were inoculated with L. monocytogenes, coated with alginate, gelatin or starch with or without the antimicrobials and stored frozen at ?18°C for 12 months. Every 2 months, samples were removed from the freezer and kept at 4°C for 30 days. Prior frozen storage at ?18°C substantially enhanced the antilisterial efficacy of the edible coatings with or without antimicrobials during the subsequent refrigerated storage. Conclusions: Plain coatings with ≥2 months frozen storage and antimicrobial edible coatings represent an effective intervention to inhibit the growth of L. monocytogenes on cold‐smoked salmon. Significance and Impact of the Study: This study demonstrates the effectiveness of the conjunct application of frozen storage and edible coatings to control the growth of L. monocytogenes to enhance the microbiological safety of cold‐smoked salmon.     

Longevity of pearl millet (Pennisetum glaucum) seeds harvested at different stages of maturity

The storage potential of seeds harvested at weekly intervals after controlled pollination was studied in three diverse cytoplasmic male sterile pearl millet (Pennisetum glaucum) lines. In the first experiment in 1989, a comparison of p₅₀ (time for viability to decline to 50% during storage) among seeds of the line DSA 105A harvested 14, 21, 28, 35 and 42 days after pollination (DAP), and then stored at 35°C with 15% moisture content or 40°C with 13% moisture content, showed that those harvested 35 DAP had the greatest longevity. In the second experiment in 1990, a comparison of p₅₀ within the lines 5141A and L 67A harvested 28, 35 and 42 DAP, and then stored at 40°C with 13% moisture content, showed that seeds of both lines harvested 42 DAP had the greatest longevity. In both the seasons, and in all three lines, maximum seed longevity (p₅₀) was attained one week after physiological maturity (defined as the end of the grain filling period), which is therefore the optimum time of harvest to obtain good quality seeds for conservation.     

Effect of freezing and freeze-drying on the viability and storage of Lilium longiflorum L. and Zea mays L. pollen

The freeze-preservation of pollen is dependent on the interaction of several factors such as freezing rate, thawing rate, freeze-drying temperature and duration, storage temperature and environment and rehydration rates. Changes in any of these variables affects the others directly or indirectly.Rapid freezing of pollen at rates of approximately 200 °C/min maintains the highest degree of viable pollen in combination with rapid thawing rates of 218 °C/min. Rapid cooling and slow rewarming resulted in a substantial loss of pollen viability. This might indicate that intracellular ice crystals formed during rapid cooling perhaps grow into larger ice masses during slow rewarming or storage at temperatures above ?50 °C.The germinability of pollen freeze-dried at temperatures below ?50 °C was also prolonged over that of the controls. Germination values for unfrozen pollen stored for 30 days at 0–5 °C averaged 50% for lily and 20% for corn. Freeze-dried pollen stored for 30 days at the same temperature yielded considerably higher viability percentages for both lily and corn pollen. Drying time is an important factor, perhaps indicating that residual moisture is critical. Freeze-dried pollen can be stored at higher temperatures than frozen and control pollen. Freeze-dried material stored for five months at 0–5 °C, upon slow rehydration yielded intact grains which has average germination percentages of 25 for lily and 15 for corn. The same pollen upon rapid rehydration showed rupturing of 20–40% of the cells and practically no germination.     

Effect of Low-Temperature Storage on Diaeretiella rapae (McIntosh) (Hymenoptera: Braconidae)

We evaluated the effects of constant low-temperature storage on Diaeretiella rapae (McIntosh) (Braconidae, Aphidiinae). Diaeretiella rapae mummies were stored at 5?±?1°C for 0–36 days. The percentage of D. rapae emergence varied (100-83%) after 0–32 days of storage. After 32 days, emergence reduced to 55%. According to our Probit analysis, 50% mortality (LT50) of the population of D. rapae was reached after 40 days of storage at 5°C. Storage for up to 32 days did not negatively affect emergence and survival. Cold exposure of D. rapae for 36 days did not influence morphological malformations, sex ratio, and emergence of the F1 generation. After 4–36 days of storage, D. rapae showed a gradual decrease in emergence, longevity, reproductive capacity, and F1 sex ratio. Diaeretiella rapae can be stored for up to 24 days at 5°C, at which time the percentage of parasitism and the F1 sex ratio remain above 38% and at 0.50, respectively.     

Flow cytometric analysis from fish samples stored at low,ultra-low and cryogenic temperatures

Flow cytometry is a valuable tool in biomedical and animal sciences. However, equipment used for such analysis presents limitations at field conditions, suggesting then preservation procedures for future analysis at laboratory conditions. In this study, freezing at low (−20 °C), ultra-low (−80 °C) and cryogenic temperatures (−196 °C, i.e. liquid nitrogen) were used as preservation procedures of fish tissue. Samples were maintained in 0.9% NaCl or lysing solution, and stored at the temperatures above for 0 (fresh control), 60, 120 and 180 days of storage. After storage, the samples were thawed and proceeded to flow cytometric analysis. Storage at low temperatures (−20 °C), both in lysing and 0.9% NaCl, exhibited poor results when analyzed after 60, 120 and 180 days, showing noisy peaks, deviation in the DNA content and absence of peaks. Ultralow (−80 °C) and cryogenic (−196 °C) temperatures, both in lysing solution and 0.9% NaCl, showed good results and high quality of histograms. Both storage procedures gave similar histograms and DNA content in comparison with control group (fresh) even after 60, 120 and 180 days of storage, exhibiting the main peak at 2C content from diploid cells and a secondary peak at 4C derived from dividing cells. In conclusion, samples may be stored for 180 days at −80 °C and −196 °C in both, 0.9% NaCl or lysing solution. As cryogenic temperatures in liquid nitrogen permits indefinite storage, this procedure should be used for long-term preservation.     

Mink semen studies: I. Liquid preservation and prospect of freezing spermatozoa

Using 22 males, 41 semen samples were collected from the vagina of mink by means of a plastic tubing attached to a 1 ml syringe. Subsamples of vaginal semen were diluted in 4 different extenders, viz., tris (tris, citric acid, glycine, fructose, glycerol and egg yolk), PVP (tris extender with polyvinyl pyrrolidone and caproic acid), milk (boiled and filtered milk with glycerol) and sodium citrate. The extended semen samples were stored at 23, 5 and −196°C for varying periods and evaluated for % motile spermatozoa. In the tris extender storage for 3 days at 5°C or for 2 days at 23°C reduced the number of spermatozoa by more than 50%. When milk was used as the extender, the motility decreased from an initial value of 68% to 10% after 5 days of 5°C and to 8% after 4 days at 23°C. The PVP extender was not suitable for storage at any temperature. After being frozen at −196°C for 2 hr, the motility ranged from 3–10% in the tris extender and was zero in milk and PVP extenders. Prolonged storage for 7 days in tris extender reduced the motility to 1–7%.     

The effect of temperature on the rate of sprout growth and development within stored onion bulbs

In order to understand better the effects of storage temperature on the time to visible sprouting in stored onions, sprout growth was measured by regularly dissecting samples from bulbs stored at 1, 10, 15 or 25°C for 243 days. The dry-weight of the shoot or sprout within stored onion bulbs increased exponentially with time. The rate of increase of sprout dry weight, as well as the rate of leaf initiation by the shoot apex was faster at 17° than at 10 or 25°C, and almost zero at 1°C. The rate of loss of dry weight from storage tissue was similar at 17°C and 25°C but slower at 10°C and slower still at 1°C.     

Lung transplantation protection after warm ischemia and 1 day preservation

This study attemps to determine the role of colloid hyperosmolar solutions in the preservation of nonischemic and ischemic lungs. Four groups of animals were studied: Control fresh lung allografts and lungs stored under hypothermic storage (4–7 °C) in a modified silica gel fraction (MSGF) for 24 hr with or without warm ischemia (0, 30, 60 min) prior to storage. Fresh lungs lived an average of 14.5 days after transplantation. Stored nonischemic lungs survived an average of 11.2 days, and ischemic (30, 60 min) stored lungs remained alive for an average of 10.5 and 9.2 days. There were no significant differences in survival between the fresh and preserved allografts. Pneumonia and/or rejection occurred in 71% of all groups. MSGF appears to be a good solution for 24-hr hypothermic storage of nonischemic and ischemic lungs.     

Survivability and storage stability of Trichoderma atroviride TRS40 preserved by fluidised bed drying on various agriculture by-products

Agriculture by-products were applied to proliferate biomass of Trichoderma atroviride TRS40 in solid state fermentation (SSF) cultures. The culture media overgrown with mycelium together with conidia were preserved by fluidised bed drying at various temperatures (50°C, 60°C and 70°C) and the received biopreparations were stored for 12 months. In order to determine the suitability of TRS40 in the production of biopreparations, the influence of preservation process and storage time on their survivability was examined. The three-component mixture proved more effective in the SSF cultures, ensuring TRS40 count at 6.07?×?10^9?CFU/g?dm, which was ca. 6 times higher than in the mono-component medium. TRS40 survivability after preservation at various temperatures ranged from 40.4% to 100%, regardless of carrier type. In turn, after 12-month storage of the biopreparations produced on the three-component medium, regardless of drying temperature, the number of viable cells ranged from 2.43?×?10⁸ to 2.49?×?10^8?CFU/g?dm. Furthermore, selected parameters of growth kinetics in the Bioscreen C system were determined. The storage time of biopreparations had various effects on growth kinetic parameters. In addition, the preserved preparations based on the TRS40 retained their capability for biosynthesis of hydrolases, even after 12 months of storage.     

Persistence of <Emphasis Type="Italic">Isaria fumosorosea</Emphasis> (Hypocreales: Cordycipitaceae) SFP-198 Conidia in Corn Oil-Based Suspension

Long-term persistence of entomopathogenic fungi as biopesticides is a major requirement for successful industrialization. Corn oil carrier was superior in maintaining germination rates of Isaria fumosorosea SFP-198 conidia during exposure to 50°C for 2 h, when compared with other oils, such as soybean oil, cottonseed oil, paraffin oil, and methyl oleate. The corn oil-based conidial suspension (91.6% germination) was also better in this regard than conidial powder (28.4% germination) after 50°C for 8 h. Long-term storage stabilities of corn oil-based conidial suspension and conidial powder at 4 and 25°C for 24 months were investigated, based on the correlation of germination rate with insecticidal activity against greenhouse whiteflies, Trialeurodes vaporariorum. Viability of conidia in corn oil was more than 98.4% for up to 9 months of storage at 25°C, and followed by 23% at 21 months. However, conidial powder had only 34% viability after 3 months of storage at 25°C, after which its viability rapidly decreased. The two conidial preparations stored at 4°C had better viabilities than those at 25°C, showing the same pattern as above. These results indicate that corn oil-based conidial suspension can be used to improve conidial persistence in long-term storage and be further applied to the formulation of other thermo-susceptible biological control agents.