Survival and colonization of Escherichia coli O157:H7 on spinach leaves as affected by inoculum level and carrier, temperature and relative humidity

Aims: To determine survival and colonization of Escherichia coli O157:H7 on spinach leaves as affected by inoculum level and carrier, temperature and relative humidity (r.h.). Methods and Results: Spinach leaves were inoculated with suspensions of E. coli O157:H7 in distilled water (DW) and 0·1% peptone water (PW) and incubated at 4, 12 and 25°C and 43, 85 and 100% r.h. The number of E. coli O157:H7 on leaves (5·6 or 1·9 log CFU per leaf) inoculated using DW as a carrier medium increased significantly at 25°C and 100% r.h. within 120 h but remained constant or decreased significantly under other test conditions. E. coli O157:H7 on leaves (5·4 log CFU per leaf) inoculated using PW as a carrier increased significantly within 72 and 24 h, respectively, at 12 or 25°C and 100% r.h.; counts using a low inoculum (2·2 log CFU per leaf) increased significantly within 24 h at 25°C. Conclusions: Escherichia coli O157:H7 can colonize on spinach leaves at 12 or 25°C in a 100% r.h. environment. Organic matter in the inoculum carrier may provide protection and nutrients which enhance survival and colonization. Significance and Impact of the Study: Colonization of E. coli O157:H7 on spinach leaves as affected by organic matter in the inoculum, temperature and r.h. was determined. These observations will be useful when developing strategies to prevent growth of E. coli O157:H7 on pre‐ and postharvest spinach.     

Threshold temperature and heat unit requirements for the development of the granary weevil,Sitophilus granarius (L.)

The objective of the present investigation was to determine the effects of temperature on the granary weevil, Sitophilus granarius L., in the laboratory so that forecasting models based on heat accumulation units could be developed for the pest. Development of S. granarius reared on wheat grains was studied at three constant temperatures (15, 20 and 25?°C) and relative humidity of 70?±?5%. The developmental time was significantly decreased with the increase in temperature. The developmental threshold temperatures estimated for egg, larva, pupa and from egg to adult were 7.79, 7.31, 14.08 and 9.97?°C, respectively. Based on these thresholds, the developmental stages, respectively, needed 92.75, 393.78, 45.04 and 562.36?day degrees to complete their development. Also, about nine generations were calculated for the pest per year under Assiut prevailing conditions. It seems that temperature of 25?°C is the most favourable developmental temperature.     

Contribution to the biology of the citrus bud mite, Aceria sheldoni (Ewing) (Acarina: Eriophyidae)*

In laboratory studies mites of Aceria sheldoni were reared on citrus fruit peels, beneath coloured cellophane hoods, to cater for their thigmotaxis and sensitivity to light of particular wave lengths. Hatching was most successful at 25°C and 98% r.h. but was reduced by low humidities (35–40% r.h.), when abnormal dwarf larvae emerged. The eggs hatched in 3–14 days; the length of a generation (egg to egg) was 12–33 days. The threshold of embryonic development was 9 °C and that for completion of the life cycle, egg to egg, was 12.5 °C. The average number of eggs laid per female was six (4–8). It increased to twelve (5–19) if the mite, during its larval stages, had been fed on buds. The vitality of both the eggs and the mature mites was tested by exposure to extreme low and high temperatures (below 0 °C, 39 °C): 50% of mites died after 30 min at 30 °C; 50% died after 30 min at –15 °C or lower.     

Virus Free Garlic (Allium Sativum L.) Plants Obtained by Thermotherapy and Meristem Tip Culture

Virus infection in garlic considerably reduces yield and quality in Argentina. The production of virus free “seed” was attempted by means of thermotherapy and meristem tip culture. A hot water treatment was employed to determine the lethal temperature/time combination for clonal type (c.t.) Blanco cloves. It was established that 50°C × 20 min, 50°C × 15 min and 55°C × 5 min were the limit thermal/time combinations which garlic could withstand. Those treatments were employed followed by meristem tip culture, however, none of the successfully developed plants after culture (only 13 %) were virus-free. Hot air treatments in a growth chamber at 36°C lasting for 30, 40 and 60 days, and at 25°–32° for 30 days in a greenhouse were tested on c.t. Blanco. Cloves kept at room temperature throughout the experiment were employed as controls. In the 25°–32°C treatment, 73% of meristems produced plants and, of these, 33 % were virus free. After 30 and 40 days at 36 °C, 62 % and 67 % of the meristems developed into plantlets, of which respectively 51 % and 50 % were virus-free. Very few meristems (10 %) developed into plants when cloves had been kept at 36°C for 60 days but the resulting plantlets were all virus free. Controls produced 78 % of plants, of which 14 % were virus free. Results of hot air treatments of 36 °C for 40 days performed on c.t. Colorado, Rosado, Paraguayo, Espaol and Hilario Ascasubi were similar to those obtained with c.t. Blanco. In Espaol and Hilario Ascasubi, no virus-free plants were detected among control specimens (no thermotherapy treatment). The only virus (from up to 3 that infected the plants) that persisted in some plants after themotherapy and meristem tip culture was garlic yellow streak.     

The life – history of the peach silver mite,Aculus fockeui (Acari: Eriophyidae) in Egypt

The peach silver mite Aculus fockeui (Nalepa and Trouessart) successfully developed from egg to adult stage when reared on a nectarine or peach leaflet attached with soft lateral divided branch dipped in test tube at different constant temperatures and 70% r.h. The effect of temperature on the development, reproduction and population growth was investigated. Below 20°C all activity ceased and by 33°C the adult began to slow down and cease all activity. At least of 34% of the generation time was spent in the egg stage at 29°C. Adult longevity decreased with increasing temperature. Fecundity was highest at 29°C with 43.16 eggs per female. Life table parameters showed that the population of A. fockeui on nectarine leaves multiplied 30.04 times in a generation time of 18.49 days at 29°C, while the population on peach leaves increased 13.87 times in a generation time of 19.18 days under the same conditions.     

Effects of environmental factors on growth of lesions on field bean leaves infected by Botrytis fabae

Chocolate spot lesions increased in size only slowly when the relative humidity of the air was below 66%. Following a lag phase immediately after infection the rate of increase was linear and proportional to humidity between c. 70% and 100% r.h. Lesions on leaflets kept at 70% r.h. for 8 h and at 100% r.h. for 16 h/day increased in size at only 27% of the rate of those at continuous 100% r.h. The optimum temperature for lesion growth was between 15 and 22 °C, the minimum <4 °C and the maximum c. 30 °C. Humidity did not interact with temperature between 10 and 20 °C. Neither light intensity nor a film of water over the leaves affected lesion growth. These findings are discussed in relation to meteorological data and field observations. The possible mechanisms whereby humidity affects lesion growth did not appear to be related to CO₂ and O₂ concentrations nor to the overall water potential of the leaf. Preliminary evidence is presented for the production of phytotoxins within lesions.     

The development of Puccinia hordei on barley cv. Zephyr

Germination of uredospores of Puccinia hordei was similar on cover-slips and on the first leaves of barley seedlings (cv. Zephyr) at 100 % r.h. over the range 5–25 °C, being greatest at 20 °C. At 15, 20 and 25 °C maximum germination was attained in 6 h. No uredospores germinated on coverslips in humidities below saturation. The numbers of pustules which subsequently developed on plants incubated at 5, 10, 15 or 18 °C and 100 % r.h. for varying periods up to 24 h, were directly related to rise in temperature and length of incubation. The time from inoculation to eruption of pustules (generation time) was 6 days at 25 °C, 8 days at 20 °C, 10 days at 15 °C, 15 days at 10 °C and 60 days at 5 °C. Pustule production on inoculated plants which had been kept at 5 °C was rapidly accelerated when they were transferred to 20 °C. Data obtained at constant temperatures were used to predict generation times of the fungus in the field. The productivity of pustules, determined as weight of uredospores, was examined at 10, 15 and 20 °C. Significantly more spores were produced at 15 than at 10 °C and most were produced at 20 °C. The results are discussed in relation to those obtained by other workers and to the development of brown rust in the field.     

THE BIOLOGY OF THE RICE WEEVIL, CALANDRA ORYZAE (L.)

The length of the developmental instars of Calandra oryzae has been estimated at relative humidities of 50, 60, 70 and 80% at 21° C., at 70% at 18, 25 and 30° C., and at 80% at 18° C., by the dissection daily of samples of infested wheat-grains. The results are expressed as a median obtained by the method of probit analysis. Comparison of this method, with estimates of median and mean obtained by orthodox arithmetical methods from similar work on Rhizopertha , show that the probit method gives good estimates.
About 90% of the eggs laid are fertile. Normally only one adult will develop in a grain, all other individuals being destroyed by cannibalism. The sex ratio is unity. It was not possible to cross C. oryzae and C. granaria.
The daily oviposition rate of C. oryzae at 17, 21 and 25° C. increases with relative humidity. There is a critical point at about 60% r.h . below which egg laying declines rapidly, and mortality is high. At 100% r.h . the oviposition rates per female per day are approximately 1.3, 2.5 and 3.4 at 17, 21 and 25° C. respectively.
In experimental conditions most eggs per grain are obtained by giving isolated females one grain each, but more eggs are laid by females given more than one grain. Daily egg output is reduced by grouping females or including males. In culture, depths of grain up to 7 cm. do not discourage egg laying.     

Factors affecting sporulation and germination of Pandora nouryi (Entomophthorales: Entomophthoraecae), a pathogen of Myzus persicae (Homoptera: Aphididae)

Pandora nouryi discharged large numbers of primary conidia between 8 and 25°C from cadavers on the surface of water-agar. At 8°C conidial discharge lasted for 120 h, but most conidia were produced within 48 h when temperature was >15°C. Saturated humidity alone was not enough to allow for sporulation to occur freely and where RH?<?95%, no conidia were discharged. Light did not affect the pattern of conidial production nor the total number of conidia. Germination percentages of conidia on the surface of water-agar were 40 and 66% at 8 and 30°C, respectively, and were significantly lower than that at 15–25°C where germination was >95%. Conidia on leaves germinated well when RH?>?74%, while no germination occurred when RH?<?100% on cover slips. All eight insecticides tested entirely inhibited conidial germination at recommended doses (R), in particular, both the organophosphorus pesticides Lorsben (chlorpyrifos) and the organochlorine pesticides Thiodan (endosulfan) completely inhibited conidial germination even at 0.2R dose.     

Response of Callosobruchus maculatus (F.) to varying temperature and relative humidity under laboratory conditions

The effect of temperature and relative humidity were determined on the development of Callosobruchus maculatus (F.) on stored bean (Vigna unguiculata) seeds exposed to five temperatures (20, 25, 30, 35 and 40 °C) and six relative humidity levels (40, 50, 60, 70, 80 and 90%). Oviposition and total adult progeny responded in a curvilinear pattern to temperature and relative humidity while developmental period presented a linear response. Egg laying (117.33 ± 3.21) and adult emergence (35.00 ± 1.70) were least at 20 °C and 90% R.H. but those showed the optimal values at 30 °C and 70% R.H. The implication of these findings is that the growth attributes of C. maculatus relate with the weather variables studied in a curvilinear manner and thus quadratic equations generated could be used for the prediction of optimum temperature and relative humidity in a given area for the management of C. maculatus in stored beans.     

Laboratory observations on the effect of humidity on the persistence of infectivity of conidia of the aphid pathogen Erynia neoaphidis

Conidia of the aphid pathogen Erynia neoaphidis on detached bean leaves and glass coverslips maintained at 20°C and at humidities from 40 to 100% r.h. lost their infectivity for pea aphids at a rate dependent on the humidity. Infectivity declined most rapidly in inocula kept on leaves at 70% r.h. and persisted longest in those at 40 and 50% r. h. That of inocula on coverslips declined most rapidly at 77% r.h. and persisted longest in those kept at 40% r.h. Even after 21 days a little infectivity was retained by conidia stored at 50% r.h. on leaves and 40% r.h. on coverslips.     

Effects of temperature on infection of French bean leaves (Phaseolus vulgaris L.) by lucerne mosaic virus

The effect of temperature on the number of lesions and the time of their appearance was studied by inoculating French bean leaves (Phaseolus vulgaris L. cv. Perli?ka) with lucerne mosaic virus either 24 or 48 h before or, 24 or 48 h after they were exposed to various temperatures. The temperatures tested were 23, 25, 27, 30, 33 and 36° C. Before and after such exposures the plants were kept in a constant temperature of 25° C. By increasing the temperature before inoculation the number of lesions increased in comparison with the control. The optimal temperature for the maximum number of lesions is between 27° and 30° C. There is no significant difference between those experiments when the exposure time was 24 h or 48 h before inoculation. The same temperatures applied for 24 or 48 h after inoculation have a decreasing effect upon the number of lesions formed by LMV on French bean leaves. The decrease is 30 to 75%. In this case the first necrotic local lesions appeared 42 h after inoculation when exposed to higher temperatures above 27° C for 24 h, and 60 h after inoculation when exposed to these temperatures for 48 h. The shape of lesions varied a little in both cases as the pictures show.     

Development times and fecundity of three important arthropod pests of strawberry in the United Kingdom

The development rates and fecundity of three important pests of strawberry in the UK were determined over a range of temperatures. Development time of the strawberry tarsonemid mite, Phytonemus pallidus, from egg lay to adult, ranged from a mean of 28.4 days at 12.5°C to 8.8 days at 25°C. No nymphs developed to adult at 10°C. Females lived for up to 45 days and laid a mean of 24.3 and 28.5 eggs at 20°C and 25°C respectively. Total development time from egg lay to adult for the strawberry blossom weevil, Anthonomus rubi, ranged from a mean of 95.7 days at 10°C to 18.2 days at 25°C. Mean fecundity at 20°C was 157.6 eggs, and the oviposition period averaged 71.6 days. When nymphs were reared on strawberry, development of the European tarnished plant bug, Lygus rugulipennis, from egg lay to adult, ranged from 83.8 days at 15°C to 28.8 days at 25°C. Development times on groundsel were shorter and ranged from 65.6 to 22.2 days at 15°C and 25°C. Only two nymphs developed to adults at 10°C; no eggs hatched at that temperature. Mean fecundity at 20°C was 75.4 eggs, but ranged from 23 to 179. Under a fluctuating temperature regime of 10°C for 12 h:20°C for 12 h, nymphs of L. rugulipennis took 40.3 days to become adult on strawberry, and 33.4 days on groundsel. Simple linear models fitted the developmental rate ‐ constant temperature relationship well for all species, accounting for 95–98% of the total variation in observed developmental rates. Development under fluctuating temperatures illustrated the potential problem of extrapolating linear models beyond the conditions of the experiment.     

Temperature dependence for development of the whitefly predator <Emphasis Type="Italic">Clitostethus arcuatus</Emphasis> (Rossi)

This work aimed to study the biology of Clitostethus arcuatus (Rossi) (Coleoptera: Coccinellidae) under different temperatures and evaluate the optimum temperature for its mass rearing. Studies were carried out in the laboratory at four constant temperatures (15°C, 20°C, 25°C and 30°C), 75 ± 5% relative humidity and a photoperiod of 16 h light:8 h dark, in which C. arcuatus was fed ad libitum with nymphs of all instar of Aleyrodes proletella L. (Homoptera: Aleyrodidae) on Brassicae oleracea L. (var. Costata). The following biological parameters were evaluated: development time and survival rates of pre-imaginal stages, adult longevity (female and male), length of the pre-oviposition and oviposition periods, fecundity, fertility and percentage of egg hatching. Population growth parameters, the lower development threshold and the sum of effective temperatures were estimated. Temperatures ranging from 20°C to 30°C were suitable for the development of C. arcuatus, suggesting that this species is well adapted to the temperatures usually found inside greenhouses or in open fields in temperate regions. Although the intrinsic rate of natural increase and doubling time were similar at 25°C and 30°C, the temperature of 25°C was shown to be the most suitable for mass rearing and development of populations under field conditions, since the percentage of egg hatching and the accumulated survival rates of the pre-imaginal stages were the highest. Considering the estimated lower threshold for pre-imaginal development (7.9°C) and the sum of effective temperatures [293.6 degree-days (°D)], it is predicted for Ponta Delgada (Azores, Portugal) that the first adults of C. arcuatus should emerge by the first fortnight of February and that up to 12 generations per year can occur.     

Measurements of electrical leaf surface conductance reveal re-condensation of transpired water vapour on leaf surfaces

Electrical conductance ( λ ) was measured continuously and in vivo on leaf surfaces of Vicia faba and Aegopodium podagraria . λ increased with rise and decreased with fall in humidity, exhibiting a hysteresis during an applied humidity cycle [90–20–-90% relative humidity (r.h.)]. After treatment with NaNO₃ aerosols, a sudden increase in λ was observed at 73% r.h., which is close to the deliquescence point of the salt. Transpiration and electrical conductance of untreated leaves were measured simultaneously under conditions of constant r.h., while the photosynthetic photon flux density and CO₂ concentration of the air were varied to induce changes of stomatal aperture. At 35% r.h., changes of light and CO₂ level revealed a strong correlation between stomatal conductance ( g _S) and λ for Vicia faba leaves. This was also found at 90, 75, 60, 45 and 25% r.h. on the lower but not on the astomatous, upper surface of Aegopodium podagraria . The correlation between g _S and λ for stomata-bearing leaf surfaces indicates that an equilibrium exists between the ambient water vapour phase and the liquid water phase on and within the cuticle. This is modified by transpired water vapour influencing the air humidity inside the boundary layer. Our results imply re-condensation of transpired water vapour to salts on the leaf surface and its sorption to the cuticle.     

Deteriorative Changes in Pea Seeds (Pisum sativum L.) Stored in Humid or Dry Conditions

Seeds stored under various conditions showed deteriorative changesin extremely dry (1% r.h. at 10 °C) or humid (93% r.h. at25 °C) conditions after 6 weeks storage, when little orno loss of viability had taken place; no changes were detectedin intermediate conditions (45% r.h. at 10 °C). The lossof electrolytes from seeds into water increased after 3 weeksof humid storage, and subsequently dead areas developed on thecotyledons of seeds held in either humid or dry conditions.With time in storage some of the seeds in dry conditions showeda reduction in the rate of imbibition, and consequently a lowlevel of electrolyte leakage. Other seeds showed an increasein leakage following dry storage. No change in solute content(sugars, potassium, and electrolytes) was detected in seedsstored in humid conditions, suggesting that the increased electrolyteleakage was caused by an impaired ability to retain solutes.Thus increased leakage was recorded in seeds whose cotyledonscontained no dead areas as revealed by vital staining, and wastherefore attributable to changes in living cells, possiblydeterioration in cell membranes. Viability began to declineafter 6 weeks in humid storage at 25 °C and after 2 d in94% r.h. at 45 °C, but was maintained in both dry and intermediateconditions. The rate at which viability fell in humid storagewas greatly influenced by the initial condition of the seed.     

Life table parameters of an indigenous strain of Neoseiulus californicus McGregor (Acari: Phytoseiidae) when fed Tetranychus urticae Koch (Acari: Tetranychidae)

The life table of the indigenous Neoseiulus californicus was studied at different temperatures and 65 ± 5% relative humidity under conditions of 16 h light : 8 h dark (LD 16:8). The total developmental period from egg to adult varied from 3.0 to 14.0 days at 15 to 35°C. Survival to adulthood ranges from 86.21 to 93.94%, with the highest rate at 25°C. The lower threshold temperature from egg to adult stages of females and males was 10.84 and 10.72°C, respectively, and the thermal constant was 57.14 degree‐days (DD) for females and 56.18 DD for males. Total number of eggs laid by each female was the highest (70.38 eggs) at 25°C, whereas average daily fecundity was the highest (3.69 eggs/female/day) at 30°C. The net reproductive rate was the highest (48.49) at 25°C and lowest (26.18) at 30°C. Mean generation time decreased from 19.04 to 11.47 days with increasing temperature from 20 to 30°C. Both intrinsic rate of natural increase (0.284) and finite rate of increase (1.32) were maximum at 30°C. Adult longevity was the highest (42.75 days for females and 32.60 days for males) at 20°C and lowest (22.70 days for females and 15.30 days for males) at 30°C. Sex ratio was female biased and was the highest (78.08) at 25°C and lowest (70.24) at 30°C. Developmental data of five constant temperatures, temperature thresholds and thermal requirements may be used to predict the occurrence, number of generations and population dynamics of N. californicus as an important biocontrol agent of Tetranychus urticae.     

Pathogenesis of mulberry leaf spot in response to meteorological aspects of Kashmir,India

In this study, the influence of climatic factors on the development of leaf spot diseases of mulberry (Morus alba L.) in Kashmir, India was studied. It was found that the highest disease incidence and severity of Goshoerami variety was found in tree type (13.86% and 1.98%) followed by dwarf (7.53% and 1.67%) and bush (3.38% and 0.99%), respectively. Due to infection of pathogen, the relative water content and chlorophyll content of leaves were consistently decreased. The order of reduction in relative water content of infected leaves was found as: bush?>?dwarf?>?tree, while as the reduction in chlorophyll content: bush?>?tree?>?dwarf. It was also found that temperature of 25–30?°C and relative humidity of above 80% favoured the development of the disease. Rapid development of disease was found during the month of June, July and August when average temperature and relative humidity was about 25–30°C and 80%, respectively.     

Lethal effects of high temperatures on nondiapausing pupae ofBathyplectes curculionis [Hym.: Ichneumonidae]

Nondiapausing pupae ofBathyplectes curculionis (Thomson) were studied under laboratory conditions. The mortality caused by 8 temperatures between 25–48°C at 20% and 70% relative humidity was measured at 10 exposure times between 15 min-24 h. There was no significant mortality at 25°C. Between 30 and 40°C, mortality occurred from long exposures only, with lethal effects becoming greater at each increase in temperature. At 43°C mortality occurred from relatively short exposures, with 100% at 4 h. Exposure times for 50% mortality averaged 16.58 h at 38°C, 1.08 h at 43°C and 0.31 h at 48°C. A slightly higher mortality occurred at 20% relative humidity than at 70% at temperatures between 35 and 40°C. At temperatures above 43°C no effects of relative humidity were noted. Afternoon soil surface temperatures in recently cut alfalfa fields commonly exceeded 50°C during July in northern Utah.     

Effect of constant temperatures on development of the walnut husk fly,Rhagoletis completa

The developmental rates of various life stages ofRhagoletis completa Cresson (Diptera: Tephritidae) were determined in the laboratory at seven different constant temperatures: 8, 12, 16, 20, 24, 28, and 32±1°C, RH 80±10%, photoperiod L 16∶D8. Preoviposition developmental rate was fastest at 28°C (10±1 days, mean±SD) and slowest at 12°C (26±1 days). About 83% of the females deposited eggs at 20 and 24°C and only 25% oviposited at 32°C. Females laid the highest number of eggs at 24°C and the lowest at 8°C. Egg development increased with increasing temperatures up to 28°C, then declined. The fastest egg development was noticed at 28°C (55±1 h) and slowest at 8°C (389±2 h). Over 90% egg hatch was observed at temperatures between 12 and 32°C, but decreased to 73% at 8°C. Larval development was fastest also at 28°C (20±0.2 days). Over 65% pupation was recorded at 20 and 24°C, but decreased to 15% at 32°C and 12% at 8°C. Pupal development was most rapid at 24°C (53±1 days) and slowest at 8°C (162±2 days). More than 70% of adult emergence was noticed in treatments between 16 and 24°C but decreased to 20% at 8°C. Based on a linear regression model of temperature-development rate relationship, the lower developmental thresholds were determined to be 6.6, 5.3, 2.9, and 5°C for preoviposition, egg, larval, and pupal stages, respectively. Based on a non-linear developmental rate model, the upper developmental thresholds were 34°C for preoviposition, egg, and larval stages and 30°C for pupal stage.