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Apple damage, pest phenology, and factors influencing the efficacy of tebufenozide for control of obliquebanded leafroller (Lepidoptera: Tortricidae).

Fruit damage by obliquebanded leafroller, Choristoneura rosaceana (Harris), was examined on four different apple cultivars during 1997-1999 in heavily infested orchards in New York State. Inital fruit damage occurred 354+/-26 degree-days base 6 degrees C (DD6) after the first moth catch from the overwintering generation and continued to increase until after the typical spray season (approximately 1,300 DD6). In addition to apple damage by late instars, fruit damage occurred by early instars of the summer and overwintering generations. The insect growth regulator tebufenozide was used as a model insecticide to determine how a slow-acting insecticide and its relative toxicity to early (neonate) and late (fourth and fifth) instars may influence the efficacy of sprays for the control of the obliquebanded leafroller. Because tebufenozide is a slow-acting compound, bioassays were conducted to determine what percentage of the total mortality to neonates occurs at each 24 h interval until 10 d. Based on a polynomial regression, half of the total mortality to larvae at the LC25, LC50, LL90 and LC99 occurred at 7.2, 5.0, 4.1, and 3.0 d, respectively. Late instars were three times more tolerant to tebufenozide than neonates.

Animals↗

Parasitism of obliquebanded leafroller (Lepidoptera: Tortricidae) in commercially managed Michigan apple orchards.

The obliquebanded leafroller, Choristoneura rosaceana (Harris), is one of the major arthropod pests of apple in the United States. In 1999 and 2000, a survey of the obliquebanded leafroller parasitoid complex in commercially managed apple orchards in Michigan's two largest fruit production regions was conducted to determine the species present and their importance to obliquebanded leafroller population management. In total, 8,961 obliquebanded leafroller larvae were collected of which 2,174 were parasitized. Parasitism increased from the overwintering generation to the summer generation for both regions and both years. In 1999, 11% of the 1,126 overwintering obliquebanded leafrollers collected were parasitized, whereas 28% of the 3,669 summer generation were parasitized. In 2000, 8% of the 489 overwintering obliquebanded leafrollers collected were parasitized, whereas 26% of the 3,677 summer generation obliquebanded leafrollers collected were parasitized. In total, 20 species of hymenopteran and dipteran parasitoids from seven families were recovered from obliquebanded leafroller larvae over the course of the study. The most abundant hymenopteran parasitoids were Bassus dimidiator Nees (Braconidae) comprising 48% of the total parasitoids, followed by Colpoclypeus florus (Walker) (Eulophidae) (8% of the total) and Macrocentrus linearis (Nees) (Braconidae) (2% of the total). Dipteran parasitoids (Tachinidae) accounted for 36% of the parasitism and were largely comprised of Nilea erecta (Coquillett) (5%) and Actia interrupta Curran (13%). These collections represent new host records for B. dimidiator, Bassus annulipes (Cresson) Hyphantrophaga blanda (Osten Sacken), and Compsilura concinnata (Meigen). The parasitoid C. florus is also reported from Michigan for the first time.

Agriculture↗

Instar-specific phenology of Pandemis pyrusana and Choristoneura rosaceana (Lepidoptera: Tortricidae) in Washington apple orchards.

Head capsule width was used to determine the instar specific phenology of the leafroller Pandemis pyrusana Kearfott and the obliquebanded leafroller, Choristoneura rosaceana (Harris) (Lepidoptera: Tortricidae), attacking apple in Washington state during 2001-2003. In total, 7012 P. pyrusana and 6122 obliquebanded leafroller larvae were measured from apple orchards from mid-March to mid-September. Degree-day accumulations from each site were paired with the head capsule data to determine the periods during which different instars were present in the field. The implications of this work for pest management and biological control of leafrollers is discussed.

Animals↗

Factors promoting infestation of newly planted, nonbearing apple orchards by dogwood borer (Lepidoptera: Sesiidae).

The initiation and level of infestation by dogwood borer, Synanthedon scitula (Harris), was tracked over three consecutive years in two nonbearing apple (Malus spp.) orchards in West Virginia and Virginia. The orchards were planted on a number of rootstock-variety (approximately cultivar) combinations and grown using different cultural practices. Infestations were detected during the first season after planting and continued to increase annually. The amount of burr knot tissue had the greatest impact on dogwood borer populations, because increasing amounts of burr knot tissue resulted in higher infestation rates. The use of plastic spiral wrap tree guards seemed to increase the development of burr knot tissue, resulting in significantly greater infestation compared with trees without tree guards in the West Virginia orchard. Variety also had a significant effect, because 'Idared' trees on M.26 had significantly greater levels of infestation compared with 'Buckeye Gala' on M.26, with or without tree guards, in the Virginia orchard. Mounding soil around the rootstock to a height just above the graft union prevented or tremendously curtailed infestation by dogwood borer, but it led to scion rooting that seemed to have an impact on size-controlling features of dwarfing rootstocks. Removal of the mounds at the beginning of the third growing season resulted in infestation of the rooted tissue during the same season. As long as apple cultivars continue to be planted on size-controlling rootstocks, dogwood borer will likely remain a serious pest, requiring either chemical treatments or a behavioral control strategy, such as mating disruption, to protect trees from infestation and damage.

Agriculture↗

Sublethal effects of chronic exposure to tebufenozide on the development, survival, and reproduction of the tufted apple bud moth (Lepidoptera: Tortricidae).

The lethal and sublethal effects of tebufenozide on the survival, development, and reproduction of a field strain of tufted apple bud moth, Platynota idaeusalis (Walker) (Lepidoptera: Tortricidae), were assessed by feeding first and third instars tebufenozide-treated diet until pupation. Larval mortality was 27.4 and 44.7% at 0.1 and 0.2 ppm for first instars and 21.9 and 57.8% at 0.2 and 0.4 ppm for third instars, respectively. Treated larvae exhibited higher pupal mortalities, lower pupal weights, and generally more deformed adults than untreated larvae. Larval development was not affected by tebufenozide when neonates were exposed, but development was accelerated slightly at 0.4 ppm for both males and females when third instars were exposed. All treatments produced sex ratios biased toward males. When paired with either treated or untreated males, females resulting from neonates treated at 0.2 ppm and from third instars treated at both 0.2 and 0.4 ppm laid from 37 to 65% fewer eggs. A reduction in fertility was only found when third instars were treated at the higher 0.4 ppm rate. These results suggest that tebufenozide can exhibit a significant effect on the population dynamics of the tufted apple bud moth.

Animals↗

Confirmation and efficacy tests against codling moth and oriental fruit moth in apples using combination heat and controlled atmosphere treatments.

Codling moth, Cydia pomonella (L.), and oriental fruit moth, Grapholita molesta (Busck), are serious pests of apples (Malus spp.) grown in the United States and other countries. In countries where these species are not found, there are strict quarantine restrictions in place to prevent their accidental introduction. The treatment used in this study consisted of hot, forced, moist air with a linear heating rate of 12 degrees C/h to a final chamber temperature of 46 degrees C under a 1% oxygen and 15% carbon dioxide environment. We found that the fourth instar of both species was the most tolerant to the treatment, with equal tolerance between the species. Efficacy tests against the fourth instar of both oriental fruit moth and codling moth by using a commercial controlled atmosphere temperature treatment system chamber resulted in > 5,000 individuals of each species being controlled using the combination treatment. Confirmation tests against codling moth resulted in mortality of > 30,000 fourth instars. These treatments may be used to meet quarantine restrictions for organic apples where fumigation with methyl bromide is not desirable.

Animals↗

Effects of apple orchard management strategies on the great tit (Parus major) in southeastern France.

The impact of conventional, organic, and integrated pest management (IPM) strategies of apple orchards on the reproduction of the great tit Parus major was investigated during a three-year period in southeastern France. The colonization process, egg-laying dates, clutch sizes, and fledging success were similar among pairs of P. major nesting in orchards conducted under the three studied management strategies. However, the mean number of young produced per ha (orchard productivity) was significantly higher in organic orchards than in both conventional and IPM orchards. Such divergences between both fledging success and orchard productivity primarily resulted from higher densities of P. major nesting pairs, but also from lower rates of nest abandonment during incubation in organic orchards. We suggest that intensive pesticide use under both IPM and conventional managements may have resulted in a substantial reduction in insect prey availability that enhanced intraspecific competition, which then led to failure in reproduction in pairs with low competitive ability. Our results highlight the relevance of P. major in assessing the environmental impact of apple orchard management strategies.

Agriculture↗

Apple- and hop-polyphenols protect periodontal ligament cells stimulated with enamel matrix derivative from Porphyromonas gingivalis.

BACKGROUND: Enamel matrix derivative (EMD) is a tissue regenerative agent used clinically as an adjunct to periodontal surgery. It was previously demonstrated that Porphyromonas gingivalis, a periodontal pathogen, significantly diminished the efficacy of EMD with periodontal ligament (PDL) cells through the proteolytic actions of Arg- and Lys-gingipains (Rgp and Kgp). Thus, antiproteolytic supplements are considered clinically desirable for effective periodontal regenerative therapies. In the present study, we examined apple- (AP) and hop-polyphenols to determine their ability to protect EMD-stimulated PDL cells from P. gingivalis. METHODS: AP, apple condensed tannin (ACT), hop bract polyphenol (HBP), high and low molecular weight fractions of HBP (HMW-HBP and LMW-HBP), and epigallocatechin gallate (EGCg) were used. PDL cells were grown on EMD-coated dishes and infected with P. gingivalis, and cellular migration and proliferation were evaluated with an in vitro assay of wound healing assay in the presence or absence of the polyphenols. RESULTS: Each polyphenol significantly enhanced the viability of PDL cells infected with P. gingivalis, whereas only EGCg demonstrated cytotoxicity. Further, all polyphenols significantly inhibited Rgp activity, with AP, ACT, and HBP more effective toward Kgp. P. gingivalis markedly diminished the migration and proliferation of EMD-stimulated PDL cells, whereas the addition of AP, ACT, HBP, and HMW-HBP significantly protected the cells from bacterial cytotoxicity. In contrast, EGCg and LMW-HBP did not show protective effects. CONCLUSION: These results suggest that AP, ACT, AP, HBP, and HMW-HBP protect EMD-stimulated PDL cells from P. gingivalis and may be therapeutically useful supplements for EMD therapy.

Adhesins, Bacterial↗

Cryptosporidiosis associated with ozonated apple cider.

We linked an outbreak of cryptosporidiosis to ozonated apple cider by using molecular and epidemiologic methods. Because ozonation was insufficient in preventing this outbreak, its use in rendering apple cider safe for drinking is questioned.

Adolescent↗

Apple juice concentrate prevents oxidative damage and impaired maze performance in aged mice.

Oxidative stress contributes to age-related cognitive decline. In some instances, consumption of fruits and vegetables rich in antioxidant can provide superior protection than supplementation with purified antioxidants. Our prior studies have shown that supplementation with apple juice concentrate (AJC) alleviates oxidative damage and cognitive decline in a transgenic murine model compromised in endogenous antioxidant potential when challenged with a vitamin-deficient, oxidative stress-promoting diet. Herein, we demonstrate that AJC, administered in drinking water, is neuroprotective in normal, aged mice. Normal mice aged either 9-10 months or 2-2.5 years were maintained for 1 month on a complete diet or a diet lacking folate and vitamin E and containing iron as a pro-oxidant, after which oxidative damage was assayed by thiobarbituric acid-reactive substances and cognitive decline as assayed by performance in a standard Y-maze. Mice 9-12 months of age were unaffected by the deficient diet, while older mice demonstrated statistically-increased oxidative damage and poorer performance in a Y maze test. Supplementation with AJC prevented these neurodegenerative effects. These data are consistent with normal aged individuals being susceptible to neurodegeneration following dietary compromise such as folate deficiency, and a hastened onset of neurodegeneration in those individuals harboring a genetic risk factor such as ApoE deficiency. These findings also support the efficacy of antioxidant supplementation, including consumption of antioxidant-rich foods such as apples, in preventing the decline in cognitive performance that accompanies normal aging.

Age Factors↗

Apple juice concentrate maintains acetylcholine levels following dietary compromise.

Oxidative stress contributes to age-related cognitive decline. In some instances, consumption of fruits and vegetables rich in antioxidant can provide superior protection than supplementation with purified antioxidants. Our prior studies have shown that supplementation with apple juice concentrate (AJC) alleviates oxidative damage and cognitive decline in adult (9-12 months) mice lacking ApoE (as a model of increased oxidative stress) and in normal aged (2-2.5 years) mice when challenged with a vitamin-deficient, oxidative stress-promoting diet. Here, we demonstrate that AJC, administered in drinking water, maintains acetylcholine levels that otherwise decline when adult and aged mice are maintained on the above deficient diet. Normal mice aged either 9-10 months or 2-2.5 years and ApoE-/- mice aged 9-10 months were maintained for 1 month on a complete diet or a diet lacking folate and vitamin E and containing iron as a pro-oxidant, and additional groups received 0.5% AJC ad libitum in drinking water. Spectrophotometric assay of acetylcholine levels revealed a significant decline in homogenates of combined frontal cortex and hippocampus for all mice maintained on the deficient diet, and a prevention of this decline in mice maintained on the deficient diet when supplemented with AJC. These findings provide a likely mechanism by which consumption of antioxidant-rich foods such as apples can prevent the decline in cognitive performance that accompanies dietary and genetic deficiencies and aging.

Acetylcholine↗

A quantification and confirmation method of patulin in apple juice by GC/MS.

A sensitive and selective method for quantification and confirmation of patulin in apple juice by GC/MS was developed. By this method, patulin was precisely determined and confirmed down to the level of 1 and 5 microg/kg in samples, respectively. Patulin was extracted with ethyl acetate from a sample and then hexane was added to the concentrated extract solution. Significant amounts of insoluble impurities were filtered off, followed by further clean-up by solid-phase extraction with combined silica gel and Florisil cartridges. The filtration step in a low-polarity condition was very effective to remove the impurities in the sample extract solution. The eluate from the cartridges was evaporated to dryness under reduced pressure and patulin was determined and confirmed by GC/MS after derivatization with 2.5% N,O-bis(trimethylsilyl)trifluoroacetamide ethyl acetate solution. Patulin was determined in the selected ion monitoring mode (m/z 226) and confirmed in the SCAN mode (m/z 40-340). The recovery from apple juice spiked with 10-500 microg/kg ranged from 93.4 to 100%. The limits of detection and quantification were 0.1 (S/N = 3) and 1 microg/kg (S/N = 30) of patulin in samples, respectively. Levels down to 5 microg/kg of patulin in sample were readily confirmed.

Factor VIII↗

Heat resistance of Escherichia coli O157:H7 in apple juice.

The objective was to determine the effect of cider composition on the heat resistance of Escherichia coli O157:H7. The average D52 value in a model Empire apple juice was 18 min with a z value of 4.8 degrees C. Increasing the Brix from 11.8 to 16.5 degrees had no effect on thermal resistance, while increasing L-malic acid from 0.2 to 0.8%, or reducing the pH from 4.4 to 3.6 sensitized the cells to heat. The greatest effect on heat resistance was afforded by the preservatives benzoic and sorbic acids: D50 values in ciders containing 1,000 mg/l were 5.2 min in the presence of sorbic acid and only 0.64 min in the presence of benzoic acid. Commercial apple juice concentrates yielded lower numbers of survivors than single-strength juices even though their higher sugar concentrations of about 46 degrees Brix increased heat resistance.

Benzoic Acid↗

Comparison of methods for determining the presence of Escherichia coli O157:H7 in apple juice.

Six methods were compared for detection of three strains of Escherichia coli O157:H7 in enrichments of inoculated apple juice. Juice was inoculated at levels varying from 0.1 to 100 CFU/ml and centrifuged after overnight storage at 4 degrees C, and pellets were incubated at 37 degrees C in nonselective enrichment broth. At hourly intervals between 5 and 10 h and at 24 h, the enrichments were tested for E. coli O157:H7 by direct fluorescent antibody (DFA), antibody-direct epifluorescent filter technique (Ab-DEFT), direct selective plating on sorbitol MacConkey agar (SMA), immunomagnetic separation coupled to either selective plating (IMS-SMA) or the polymerase chain reaction (IMS-PCR), and flow cytometry (FC). The most consistent detection of 0.1 CFU/ml of the slowest growing strain of the pathogen was provided by the IMS-SMA and IMS-PCR after 8 h of enrichment. The time required for detection at the level of 0.1 CFU/ml for each assay was Ab-DEFT, 11 h; IMS-PCR, 16 h; FC, 24 h; IMS-SMA, 32 h; and SMA, 48 h. Absolute detection limits (without enrichment) were: IMS-PCR, 10(3) CFU/ml; Ab-DEFT and IMS-SMA, 10(4) CFU/ml; SMA, 10(5) CFU/ml; and DFA, 10(6) CFU/ml. Recovery of the pathogen (10 CFU/ml) in apple juice after 28 days of 4 degrees C storage was possible by means of an 8-h enrichment and Ab-DEFT, IMS-PCR, or IMS-SMA.

Bacteriological Techniques↗

Identification of Listeria monocytogenes from unpasteurized apple juice using rapid test kits.

A microbiological survey of 50 retail juices was conducted in the fall of 1996. These juices were analyzed for Listeria monocytogenes, Escherichia coli O157:H7, Salmonella, coliforms, fecal coliforms, and pH. Two unpasteurized juices were positive for L. monocytogenes: an apple juice and an apple raspberry blend with a pH of 3.78 and 3.75, respectively. Three L. monocytogenes isolates were characterized. The colonies were typical for Listeria sp. on Oxford and lithium chloride-phenylethanol-moxalactam agars and were beta-hemolytic on sheep blood agar. The isolates required 5 days of incubation at 35 degrees C to produce a positive rhamnose reaction in a phenol red carbohydrate broth. This slow rhamnose utilization resulted in these isolates not being identified using the Micro-ID test strip (Organon Technika). However, the isolates were positive for L. monocytogenes using the API Listeria strip (BioMerieux) and a multiplex polymerase chain reaction for detection of the hemolysis (hyla) and invasion-associated protein (iap) genes.

Bacteriological Techniques↗

Validation of apple cider pasteurization treatments against Escherichia coli O157:H7, Salmonella, and Listeria monocytogenes.

Time and temperature pasteurization conditions common in the Wisconsin cider industry were validated using a six-strain cocktail of Escherichia coli O157:H7 and acid-adapted E. coli O157:H7 in pH- and degrees Brix-adjusted apple cider. Strains employed were linked to outbreaks (ATCC 43894 and 43895, C7927, and USDA-FSIS-380-94) or strains engineered to contain the gene for green fluorescent protein (pGFP ATCC 43894 and pGFP ATCC 43889) for differential enumeration. Survival of Salmonella spp. (CDC 0778. CDC F2833, and CDC H0662) and Listeria monocytogenes (H0222, F8027, and F8369) was also evaluated. Inoculated cider of pH 3.3 or 4.1 and 11 or 14 degrees Brix was heated under conditions ranging from 60 degrees C for 14 s to 71.1 degrees C for 14 s. A 5-log reduction of nonadapted and acid-adapted E. coli O157:H7 was obtained at 68.1 degrees C for 14 s. Lower temperatures, or less time at 68.1 degrees C, did not ensure a 5-log reduction in E. coli O157:H7. A 5-log reduction was obtained at 65.6 degrees C for 14 s for Salmonella spp. L. monocytogenes survived 68.1 degrees C for 14 s, but survivors died in cider within 24 h at 4 degrees C. Laboratory results were validated with a surrogate E coli using a bench-top plate heat-exchange pasteurizer. Results were further validated using fresh unpasteurized commercial ciders. Consumer acceptance of cider pasteurized at 68.1 degrees C for 14 s (Wisconsin recommendations) and at 71.1 degrees C for 6 s (New York recommendations) was not significantly different. Hence, we conclude that 68.1 degrees C for 14 s is a validated treatment for ensuring adequate destruction of E. coli O157:H7, Salmonella spp., and L. monocytogenes in apple cider.

Adaptation, Physiological↗

Modeling the survival of Escherichia coli O157:H7 in apple cider using probability distribution functions for quantitative risk assessment.

Outbreaks of foodborne illness from apple cider have prompted research on the survival of Escherichia coli O157:H7 in this food. Published results vary widely, potentially due to differences in E. coli O157:H7 strains, enumeration media, and other experimental considerations. We developed probability distribution functions for the change in concentration of E. coli O157:H7 (log CFU/day) in cider using data from scientific publications for use in a quantitative risk assessment. Six storage conditions (refrigeration [4 to 5 degrees C]; temperature abuse [6 to 10 degrees C]; room temperature [20 to 25 degrees C]; refrigerated with 0.1% sodium benzoate, 0.1% potassium sorbate, or both) were modeled. E. coli survival rate data for all three unpreserved cider storage conditions were highly peaked, and these data were fit to logistic distributions: ideal refrigeration, logistic (-0.061, 0.13); temperature abuse, logistic (-0.0982, 0.23); room temperature, logistic (-0.1, 0.29) and uniform (-4.3, -1.8), to model the very small chance of extremely high log CFU reductions. There were fewer published studies on refrigerated, preserved cider, and these smaller data sets were modeled with beta (4.27, 2.37) x 2.2 - 1.6, normal (-0.2, 0.13), and gamma (1.45, 0.6) distributions, respectively. Simulations were run to show the effect of storage on E. coli O157:H7 during the shelf life of apple cider. Under every storage condition, with and without preservatives, there was an overall decline in E. coli O157:H7 populations in cider, although a small fraction of the time a slight increase was seen.

Beverages↗

Phosphate buffer increases recovery of Escherichia coli O157:H7 from frozen apple juice.

It is common practice to dilute food products in 0.1% peptone before microbiological analysis. However, this diluent may not be appropriate for detection of injured organisms present in acidic foods. Shelf-stable unclarified apple juice (pH 3.6) was inoculated with approximately 1 x 10(7) CFU/ml of Escherichia coli O157:H7 and held at 23 +/- 2 degrees C (control) or frozen to -20 +/- 2 degrees C for 24 h to induce injury before sampling. Unfrozen or frozen and thawed juice was diluted 1:1 or 1:10 in 0.1% (wt/vol) peptone (pH 6.1) or 0.1 M phosphate buffer (pH 7.2). Juice samples were plated onto tryptic soy agar with 0.1% (wt/vol) sodium pyruvate (TSAP) to measure survival or onto sorbitol MacConkey agar (SMA) to indicate injury. Counts on TSAP or SMA were the same for control samples held in peptone or phosphate buffer for up to 45 min. However, populations of E. coli in frozen and thawed samples declined rapidly upon dilution in 0.1% peptone. Within 20 min, E. coli underwent a >1-log10 CFU/ml reduction in viability as measured on TSAP and a >2-log10 CFU/ml reduction to below the limit of detection (1.6 or 2.3 log10 CFU/ml) on SMA. In contrast, populations of E. coli in frozen and thawed samples diluted in phosphate buffer did not decrease significantly on TSAP and decreased by <0.6 log CFU/ml on SMA during a 45-min holding period. The acidity of apple juice appears to interfere with the recovery of freeze-thaw-injured E. coli O157:H7 during sampling. Using 0.1 M phosphate buffer (pH 7.2) as a diluent results in superior recovery of these organisms on both selective and nonselective plating media.

Beverages↗