Dry mycelium preparations of entomopathogenic fungi,Metarhizium anisopliae and Beauveria bassiana

Several different harvesting procedures were used to obtain dry mycelium preparations of the entomopathogenic fungi, Metarhizium anisopliae and Beauveria bassiana. The effects of these procedures on the survival of the fungal preparations and on their conidiation after short periods of storage at room temperature and at 4°C were examined. Harvesting procedures consisted of filtering the mycelium produced in airlift containers from the culture medium, washing with deionized water, spraying with a sugar solution, and incubating for 18 hr at 4°C before drying. Conidial production of treated mycelia stored 1.5 and 4.5 months at 4°C was not significantly different for and procedure. For dry mycelium of M. anisopliae stored 1.5 months at 4°C and then at room temperature for 3 months, maltose- and sucrose-treated preparations produced more conidia than preparations sprayed with dextrose solution, with water only, or not sprayed. B. bassiana preparations dried soon after mat formation were superior to those incubated at 4°C, and maltose-and dextrose-treated mycelia were superior to other treatments when stored at room temperature.     

Changes in germinability,lipid peroxidation,and antioxidant enzyme activities in pear stock (<Emphasis Type="Italic">Pyrus betulaefolia</Emphasis> Bge.) seeds during room- and low-temperature storage

The aim of this study was to determine if loss of germinability in Pyrus betulaefolia seeds stored at 4°C and at room temperature is associated with a loss of membrane lipid peroxidation or changes in antioxidant enzyme activities. The results indicated that germination percentage clearly decreased when seeds were stored at room temperature rather than at 4°C from 6 to 12 months. Room-temperature storage of the pear stock seed for 12 months decreased germination to 15.52%, but germination percentage was not changed when seed was stored at 4°C for 12 months. MDA, a marker for membrane lipid peroxidation, increased significantly under room-temperature storage conditions. Antioxidant enzyme (SOD, POD, and CAT) activities were a good indicator of germination percentage in pear stock seeds. Antioxidant enzyme activities of pear stock seeds at 4°C were higher than antioxidant enzyme activities in seeds stored at room temperature from 6 to 12 months. Antioxidant enzyme activities of the pear stock seed decreased markedly under conditions of room-temperature storage from 6 to 12 months. The results of this study showed that long-term room-temperature storage was detrimental for maintaining the vigor of P. betulaefolia seeds. The mechanisms responsible for this outcome are a higher level of membrane lipid peroxidation and a lower level of activity of antioxidant enzymes.     

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.     

Low temperature storage effects on two olive fruit fly parasitoids

Effects of cold storage temperatures and storage duration were evaluated for Psyttalia humilis (Silvestri) from Namibia and Psyttalia ponerophaga (Silvestri) from Pakistan, braconid parasitoids of Bactrocera oleae (Rossi) imported to California, USA. Immature stages of P. humilis were exposed to 4, 6, 8, 10, or 12 °C for 1, 2 or 4 months (pupa only at 4 and 12 °C) and then held at 24 °C for adult emergence. Less than 5 % of parasitoids in the 4–8 °C treatments survived, regardless of storage duration. At the 10 °C treatment, adult survival decreased with increased storage duration, but increased with advancing developmental stages. Survival was not affected at the 12 °C treatment. Adult P. humilis were exposed to 6, 8, 10 °C for short periods (1, 2, 4, or 6 weeks) or ambient winter conditions in Parlier, California, USA (about 9 °C). Regardless of storage temperature, P. humilis reproduction was reduced after storage of four and six weeks. Similarly, after 4 months at ambient winter temperatures, P. humilis reproduction was reduced. Psyttalia ponerophaga pupae stored at 6 °C for 41–97 days had decreased survival and increased developmental time. Survival of P. ponerophaga pupae ranged from 13.9–52.1 %, whereas under similar storage conditions survival of P. humilis was <0.7 %, suggesting P. ponerophaga is more cold tolerant than P. humilis.     

Mass production and storage of Vairimorpha necatrix (protozoa: Microsporida)

Mass production and storage methods were evaluated for maximization of spores of Vairimorpha necatrix, a promising protozoan for microbial control due to its virulence and prolificity in lepidopterous pests. In vivo spore production was at a maximum when 3rd instar Heliothis zea were exposed to 6.6 spores/mm² of artificial diet surface and reared for 15 days. Approximately 1.67 × 10¹⁰ spores/larva were produced, or ca. 1 × 10¹⁰ spores/larva after partial purification of the spores by homogenization of the larvae in water, filtration, and centrifugation. The spores were inactivated by relatively short exposures to several chemicals which were tested to counteract contamination of the diet surface by fungi in the spore inoculum. Spores of V. necatrix were stored at refrigerated and freezing temperatures for up to 2 years and bioassayed periodically with 2nd instar H. zea. Spores lost little infectivity after 23 months at 6°C if they were stored in a purified water suspension plus antibiotic, but they were noninfective after 18 months at 6°C if stored in host tissue. Storage at ?15°C caused little loss of infectivity whether the spores were stored in water and glycerine, in host tissue, or after lyophilization. The spores withstood lyophilization in host cadavers better than in purified water suspension. Samples of a dry V. necatrix-corn meal formulation, which was prepared for field efficacy tests and stored at ?15° and 6°C, were highly infective after 9 months. Large numbers of V. necatrix spores can thus be produced and later made available for microbial control field trials with little loss of infectivity.     

Effects of delaying fungicide treatment of wounded potatoes on the incidence of Fusarium dry rot in store

Potato tubers artificially inoculated with Fusarium solani var. coeruleum or F. sulphureum 3 months after harvest were uniformly wounded and held at 5, 10 or 15°C for up to 21 days before immersion in fungicide suspensions. Holding tubers for 14 days at 15°C (curing conditions) or at 5°C did not alter the incidence of dry rot subsequently developing on tubers stored at 10°C, and holding tubers for up to 21 days at 15°C slightly increased disease caused by both pathogens. Thiabendazole, imazalil and prochloraz applied to tubers immediately after wounding almost completely prevented dry rot. Treatment after 3 days was less effective and the amount of disease increased with further delay; fungicides were more effective on tubers held at 5°C than at 10 or 15°C before treatment and storage, and efficacy of the fungicide was decreased by increasing the amount of inoculum on tubers. Wounds became less susceptible to infection by F. solani var. coeruleum and F. sulphureum when tubers were held at 15°C before inoculation, and the incidence of rots was decreased by 70–80% by delaying inoculation for 7 days. Treating tubers with dichlorophen immediately after wounding slightly increased the disease. The effects of fungicide treatment, curing conditions and wound healing are discussed.     

Does vacuum-packaging or co-delivered amendments enhance shelf-life of Striga-mycoherbicidal products containing Fusarium oxysporum f. sp. strigae during storage?

The effect of vacuum packaging on the shelf-life and handling of Pesta granules and seed treatment made with chlamydospores of Fusarium oxysporum strains Foxy2, PSM197 or their mixture was studied at 4°C and 22±3°C over 1 year. In addition, the effects of co-incorporated amendments [urea in Pesta or co-delivered fungicides (Ridomil Gold®, Apron XL®) on coated sorghum seeds], and coating material (Arabic gum ‘AG’, SUET Binder ‘SB’) on the viability of Striga-mycoherbicides were evaluated. Storage under vacuum packaging did not enhance shelf-life of the formulated Striga-mycoherbicidal products after 12 months of storage regardless of the treatment used. The co-incorporated urea into Pesta granules significantly reduced the viability of mycoherbicides, but less so at 4°C (58% strain-stability after 12 months). No significant differences between the coating materials in maintaining the viability of mycoherbicides were observed. The shelf-life of isolates on coated seeds significantly decreased when adding Ridomil Gold®. However, at 4°C, the fungicide Apron XL® allowed better survival of Foxy2 and PSM197 by maintaining their averaged half-lives (t _0.5) by an additional 6 months compared to Ridomil Gold®. In general, Striga-mycoherbicidal product combinations exhibited a significantly higher shelf-life when stored at 4°C than at 22±3°C. The absence of a positive effect of vacuum packaging on shelf-life of Striga-mycoherbicidal products reflects the tolerance of the formulated fungal propagules (chlamydospores) to withstand an oxygen enriched environment and allows their handling and distribution through ordinary packaging systems in Africa. The high compatibility between Striga-mycoherbicides and the co-delivered fungicide Apron XL®, and the fungal storage stability allows simultaneous control of Striga and fungal cereal diseases within an integrated pest management (IPM).     

Preservation of coagulation efficiency ofMoringa oleifera, a natural coagulant

In recent years, there has been an interest to useMoringa oleifera as the natural coagulant due to cost, associated health and environmental concerns of synthetic organic polymers and inorganic chemicals. However, it is known thatM. oleifera as the natural coagulant is highly biodegradable and has a very short shelf life. This research was carried out to investigate the effects of storage temperature, packaging methods, and freeze-drying on the preservation ofM. oleifera seeds powders. Non freeze-driedM. oleifera was prepared into different packaging namely open container, closed container and vacuum packing, whilst, freeze-driedM. oleifera was stored in closed container and vacuum packing. Each of the packaging was stored at room temperature (30 to 32°C) and refrigerator (4°C). The turbidity removal efficiencies of storedM. oleifera were examined using jar test at monthly interval for 12 months. The results indicated that non freeze-driedM. oleifera kept in the refrigerator (4°C) would preserve its coagulation efficiency. In addition, closed container and vacuum packing were found to be more appropriate for the preservation of non freeze-driedM. oleifera, compared to open container. Freeze-driedM. oleifera retained its high coagulation efficiency regardless the storage temperature and packaging method for up to 11 months. Besides, higher increment in zeta potential values for water coagulated with freeze-driedM. oleifera indicated the higher frequency of charge neutralization and better coagulation efficiency of freeze-driedM. oleifera, compared to non freeze-dried seeds. As a coagulant,M. oleifera did not affect the pH of the water after treatment.     

Seed storage and germination in Kumara plicatilis,a tree aloe endemic to mountain fynbos in the Boland,south-western Cape,South Africa

Seed storage under appropriate conditions is a relatively inexpensive means of safeguarding plant genetic material for ex situ conservation. Post-storage germination trials are used to determine the viability of stored seeds, and hence the efficacy of the particular storage treatment. Kumara plicatilis (= Aloe plicatilis) is a tree aloe endemic to mountain fynbos in the Boland, south-western Cape. The viability and germination behaviour of K. plicatilis seeds were assessed for seeds stored for four and nine months at − 80 °C, 4 °C, 25 °C and under ambient conditions in a laboratory. Seeds were germinated under controlled conditions and germination rates and percentages determined. Ungerminated seeds were tested for viability using tetrazolium salt. Seed viability was not significantly reduced during storage. Seeds stored at − 80 °C for four and nine months exhibited the fastest germination rate overall (both 5.9 ± 0.3 weeks, mean ± S.E.), and slowest was for seeds stored under ambient conditions for four and nine months (both 7.8 ± 0.4 weeks). All seed lots showed similar percentage germination after four months of storage (78.0–90.4%). The highest percentage germination overall was for seeds stored at − 80 °C for four months (90.4%) and the lowest was for seeds kept at 4 °C and − 80 °C for nine months (39.2 and 39.6%, respectively). Respective percentage viability for ungerminated seeds in these two treatments was 82% and 87%, respectively, indicating the induction of secondary dormancy. Induced dormancy triggered by protracted cold temperatures may be an adaptation that enables seeds to survive prolonged extreme conditions that are unfavourable for germination. Further research on the long-term storage of aloe seeds would be beneficial for developing long-term seed storage and germination testing protocols for ex situ conservation.     

Freezing-induced xylem cavitation and the northern limit of Larrea tridentata

We investigated the occurrence of freezing-induced cavitation in the evergreen desert shrub Larrea tridentata and compared it to co-occurring, winter-deciduous Prosopis velutina. Field measurements indicated that xylem sap in L. tridentata froze at temperatures below c. –5°C, and that this caused no measurable cavitation for minimum temperatures above –7°C. During the same period P. velutina cavitated almost completely. In the laboratory, we cooled stems of L. tridentata to temperatures ranging from –5 to –20°C, held them at temperature for 1 or 12 h, thawed the stems at a constant rate and measured cavitation by the decrease in hydraulic conductivity of stem segments. As observed in the field, freezing exotherms occurred at temperatures between –6.5 and –9°C and as long as temperatures were held above –11°C there was no change in hydraulic conductivity after thawing. However, when stems were cooled to between –11°C and –20°C, stem hydraulic conductivity decreased linearly with minimum temperature. Minimum temperatures between –16 and –20°C were sufficient to completely eliminate hydraulic conductance. Record (>20 year) minimum isotherms in this same range of temperatures corresponded closely with the northern limit of L. tridentata in the Mojave and Sonoran deserts.     

Effects of temperature, light, storage conditions, sowing time, and burial depth on the seed germination of Cardiocrinum cordatum var. glehnii (Liliaceae)

In this study, we conducted experiments to accumulate practical information on the propagation and establishment of a population of Cardiocrinum cordatum var. glehnii by seed sowing. C. cordatum var. glehnii seeds require approximately 19 months from seed dispersal to cotyledon emergence in the field. However, the period from seed dispersal to radicle emergence was shortened to approximately 7–8 months by the temperature transition of 25/15°C (60 days) → 15/5°C (30 days) → 0°C (120 days) → 15/5°C (i.e., 15/5°C represents alternating temperature treatment wherein the seeds were placed at 15°C for 12 h during the day and then at 5°C for 12 h during the night). More than 90% of the seeds, which were stored dry at 5°C for 12 months and sown in pots in the field, showed cotyledon emergence, whereas in seeds stored dry at 25°C, dry at room temperature, and non-dry at room temperature, cotyledon emergence was decreased by less than 1%. More than 88% of the seeds that were stored dry at 5°C and sown in the field in October 2002 immediately after collecting, November, and from April to July 2003 showed cotyledon emergence in spring 2004. However, seeds sown in August, September, and October 2003 showed cotyledon emergences of 57.6%, 0%, and 0% in spring 2004, respectively. Seeds collected in October 2002 and sown until July 2003 in the field received adequate high temperature in summer, moderate temperature in autumn, and cold temperature in winter; therefore, the percentage of cotyledon emergence was high in spring 2004. On the other hand, seeds sown in August 2003 or later could not receive enough high temperature; thus, cotyledons emerged from only a few seeds.     

Effect of pollination time,the hour of daytime,pollen storage temperature and duration on pollen viability,germinability, and fruit set of date palm (Phoenix dactylifera L.) cv "Deglet Nour"

Success artificial pollination with viable pollen is crucial process in the production chain of date palms. This study evaluated the impact of pollen storage temperature and duration, pollination time following spathe cracking, and the hour of daytime on pollen viability, germinability, fruit set and yield of 'Deglet Nour' date palm cultivar. In in vitro tests, fresh pollen showed the maximum viability (96.3%) and germination (85%) but it decreased thereafter upon the storage temperature (28, 4 and ?30 °C) and duration (3, 6, 9 and 12 months). In this respect, pollen stored at ?30 °C retained highest viability and germinability followed by those stored at 4 and then at 28 °C. In filed experiments, fruit set was 85, 75, 65, and 45% with pollination using fresh pollen, or pollen stored at ?30, 4 and 28 °C, respectively. Fruit set was 95%, 75%, and less than 50%, for pollination performed on the same day of spathe cracking, 6 and 12 days later, respectively. The highest fruit set percentage and yield/bunch were obtained with pollination performed between 12.0 pm and 15.0 pm in contrast to 8.0–11.0 am or 16.0–17.0.     

Reduced leaching of the herbicide MCPA after bioaugmentation with a formulated and stored Sphingobium sp.

The use of pesticides on sandy soils and on many non-agricultural areas entails a potentially high risk of water contamination. This study examined leaching of the herbicide 4-chloro-2-methylphenoxyacetic acid (MCPA) after bioaugmentation in sand with differently formulated and stored Sphingobium sp. T51 and at different soil moisture contents. Dry formulations of Sphingobium sp. T51 were achieved by either freeze drying or fluidised bed drying, with high initial cell viability of 67–85 %. Storage stability of T51 cells was related to formulation excipient/carrier and storage conditions. Bacterial viability in the fluidised bed-dried formulations stored at 25 °C under non-vacuum conditions was poor, with losses of at least 97 % within a month. The freeze-dried formulations could be stored substantially longer, with cell survival rates of 50 %, after 6 months of storage at the same temperature under partial vacuum. Formulated and long-term stored Sphingobium cells maintained their MCPA degradation efficacy and reduced MCPA leaching as efficiently as freshly cultivated cells, by at least 73 % when equal amounts of viable cells were used. The importance of soil moisture for practical field bioaugmentation techniques is discussed.     

Neutrophil migration in canines: In vivo comparison of granulocyte function following 24-hour storage at 6 or 20 °C of leukocyte concentrates or of granulocytes purified by counterflow centrifugation-elutriation

The use of granulocyte-rich concentrates from leukapheresis purified by counterflow centrifugation—elutriation to obtain pure granulocytes for transfusion studies in cyclo-phosphamide-induced neutropenic animal models is reported. Our data for granulocyterich leukapheresis concentrates indicate that room temperature (20 °C) appears to be preferred to 6 °C for short-term granulocyte storage. The data also indicate that although the granulocytes isolated by counterflow centrifugation—elutration may retain in vitro functions of chemotaxis, phagocytosis, and bactericidal activity, the in vivo function of migration into skin chambers for isolated granulocytes is seriously impaired after storage for 18 to 24 hr at both 6 and 20 °C. This loss of in vivo function of stored granulocytes occurs in isolated granulocytes obtained by both counterflow centrifugation-elutriation and dextran sedimentation, and it is not observed in the leukocyte concentrates held at 20 °C. The results of these studies are fourfold. First, freshly isolated granulocytes display no apparent loss of either in vivo or in vitro function. Second, granulocytes isolated by counterflow centrifugation-elutriation or dextran sedimentation and stored at 6 or 20 °C are severely impaired in terms of their in vivo chemotactic function but display no loss of in vitro efficacy. Third, 20 °C storage of granulocyte-rich leukapheresis concentrates for 18 to 24 hr is superior to 6 °C storage. Fourth, in vitro analysis may be limited in its ability to indicate in vivo function as a measure of success in granulocyte preservation studies.     

Determination of trovafloxacin,a new quinolone antibiotic,in biological samples by reversed-phase high-performance liquid chromatography

A simple, accurate and precise high-performance liquid chromatographic method was developed and validated for the determination of trovafloxacin, a new quinolone antibiotic, in serum and urine. Following solid-phase extraction, chromatographic separation was accomplished using a C₁₈ column with a mobile phase consisting of 0.04 M H₃PO₄-acetonitrile-tetrabutylammonium hydroxide-0.005 M dibutyl amine phosphate (D-4) reagent (83:16.85:0.05:0.1, v/v), pH 3. Trovafloxacin and the internal standard (a methyl derivative of trovafloxacin) were detected by ultraviolet absorbance at 275 nm. The lower limit of quantification for trovafloxacin was 0.1 μg/ml and the calibration curves were linear over a concentration range of 0.1 to 20..0 μg/ml (r² = 0.9997). The average recoveries were greater than 70% for both trovafloxacin and internal standard. The intra-day and inter-day coefficients of variation were generally less than 5% in urine and serum over the concentration range of 0.1 to 20.0 μg/ml. Human serum samples could be stored for up to 12 months at −20°C and urine samples could be stored up to 18 months at −80°C.     

Influence of an Acrylic Polymer Blend on the Physical Stability of Film-Coated Theophylline Pellets

The purpose of this study was to investigate the physical stability of a coating system consisting of a blend of two sustained release acrylic polymers and its influence on the drug release rate of theophylline from coated pellets. The properties of both free films and theophylline pellets coated with the polymer blend were investigated, and the miscibility was determined via differential scanning calorimetry. Eudragit^® RS 30 D was plasticized by the addition of Eudragit^® NE 30 D, and the predicted glass transition temperature (T _g) of the blend was similar to the experimental values. Sprayed films composed of a blend of Eudragit^® NE 30 D/Eudragit^® RS 30 D (1:1) showed a water vapor permeability six times greater than films containing only Eudragit^® NE 30 D. The presence of quaternary ammonium functional groups from the RS 30 D polymer increased the swellability of the films. The films prepared from the blend exhibited stable permeability values when stored for 1 month at both 25°C and 40°C, while the films which were composed of only Eudragit^® NE 30 D showed a statistically significant decrease in this parameter when stored under the same conditions. Eudragit^® NE 30 D/Eudragit^® RS 30 D (1:1)-sprayed films decreased in elongation from 180% to 40% after storage at 40°C for 1 month, while those stored at 25°C showed no change in elongation. In coated pellets, the addition of Eudragit^® RS 30 D to the Eudragit^® NE 30 D increased the theophylline release rate, and the pellets were stable when stored at 25°C for a period of up to 3 months due to maintenance of the physico-mechanical properties of the film. Pellets stored at 40°C exhibited a decrease in drug release rate over time as a result of changes in film physico-mechanical properties which were attributed to further coalescence and densification of the polymer. When the storage temperature was above the T _g of the composite, instabilities in both drug release rate and physical properties were evident. Stabilization in drug release rate from coated pellets could be correlated with the physico-mechanical stability of the film formulation when stored at temperatures below the T _g of the polymer.     

Cold storage of the egg parasitoids Trissolcus basalis (Wollaston) and Telenomus podisi Ashmead (Hymenoptera: Scelionidae)

Adults of Trissolcus basalis and Telenomus podisi were stored either at 15 or 18 °C after their immature development had been completed at 18 or 25 °C. Longevity of the parasitoids in the storage temperatures was evaluated, as well as fecundity and longevity following their return to 25 °C after different periods in reproductive diapause. Temperature during immature development influenced female longevity and highest mean longevity was obtained for females that developed to the adult stage at 25 °C and then were stored at 15 °C (ca. 13 months for T. basalis and 10 months for Te. podisi). For adults of T. basalis that developed at 25 °C, storage periods of 120 or 180 days at 15 or 18 °C did not affect fecundity. The fecundity of T. basalis females that developed at 18 °C and were stored for 120 days at 15 or 18 °C was not affected; however, after remaining for 180 days, fecundity was reduced in ca. 30 and 50%, respectively. Storage of Te. podisi adults at 15 or 18 °C significantly reduced fecundity. It is concluded that adults of T. basalis can be stored in the adult stage at 15 or 18 °C between two soybean crop seasons for mass production purposes, aiming the biological control of stink bugs.     

Effects of temperature change on the innate cellular and humoral immune responses of orange-spotted grouper Epinephelus coioides and its susceptibility to Vibrio alginolyticus

Orange-spotted grouper Epinephelus coioides held at 27 °C were then further cultured at 19, 27 (control), and 35 °C, and were examined for innate cellular and humoral responses after 3–96 h. The total leucocyte count, respiratory burst, and phagocytic activity significantly decreased 3, 48, and 96 h after fish were transferred to 19 and 35 °C. Both the alternative complement pathway (ACH₅₀) and the lysozyme activity significantly decreased at 3–96 h after fish were transferred to 19 and 35 °C. In another experiment, groupers reared at 27 °C at 34‰ salinity were injected with Vibrio alginolyticus grown in tryptic soy broth (TSB) at a dose of 2.3 × 10⁹ colony-forming units (cfu) fish^?1, and then further reared in water temperatures of 19, 27 (control), and 35 °C. The cumulative mortalities of V. alginolyticus-injected fish held in 19 and 35 °C were significantly higher than that of injected fish held in 27 °C. Resistance had decreased after 12 h for the challenged grouper held at 35 °C. All injected fish held in 19 °C had died after 72 h. It was concluded that at 12 h after transfer of grouper from 27 to 19 and 35 °C, immunity was suppressed and resistance against V. alginolyticus had decreased.     

Investigation into the Potential of Bacteriocinogenic Lactobacillus plantarum BFE 5092 for Biopreservation of Raw Turkey Meat

The bacteriocin-producing Lactobacillus plantarum BFE 5092 was assessed for its potential as a protective culture in the biopreservation of aerobically stored turkey meat. This strain produces three bacteriocins, i.e. plantaricins EF, JK and N. The absolute expression of Lactobacillus plantarum BFE 5092 16S rRNA housekeeping gene, as well as l-ldh, plnEF and plnG genes as determined by quantitative, real-time-PCR, revealed that these genes were expressed to similar levels when the strain was grown at 8 and 30 °C in MRS broth. On turkey meat, Lactobacillus plantarum BFE 5092 did not grow but survived, as indicated by similar viable cell numbers during a 9-day storage period at 8 °C. When inoculated at 1 × 10⁷ CFU/g on the turkey meat and subsequently stored at 10 °C, the culture did again not show good growth. Lactobacillus plantarum BFE 5092 could not inhibit the growth of naturally occurring listeriae or Gram-negative bacteria on the turkey meat at 10 °C, or that of Listeria monocytogenes when it was co-inoculated at a level of 1 × 10⁵ CFU/g. Gene expression analyses showed that the bacteriocin genes were expressed on turkey meat stored at 10 °C. Moreover, the investigation into the absolute expression of the three plantaricin genes of Lactobacillus plantarum BFE 5092 in co-culture with Listeria monocytogenes on turkey meat by qRT-PCR showed that the plantaricin genes were indeed expressed during the low-temperature storage condition. The Lactobacillus plantarum BFE 5092 strain overall could not effectively inhibit L. monocytogenes and therefore it would not make a suitable protective culture for biopreservation of turkey meat stored aerobically at low temperature.