Toxicity of phosphine to the diapausing stages of Ephestia elutella, Plodia interpunctella and other lepidoptera

Diapausing larvae of the stored product moths Ephestia elutella (Hübner) and Plodia interpunctella (Hübner) were tolerant of phosphine only in short exposures, the tolerance of E. elutella being at least twice that of P. interpunctella. The tolerance of a freshly collected wild stock of each species was, at some temperatures, about two or three times that in a laboratory reared stock. The wild stock of E. elutella required concentration-time (CT) products of 36 mg hr/l in 4-day exposures at 10° and 15°C, and 18 mg hr/l in 2-day exposures at 20°C and above for 100% kill. The tolerance of E. elutella larvae in diapause was not dependent on the manner in which diapause was induced, maintained or subsequently terminated. Longer exposures to phosphine were more effective than shorter ones of similar CT product because an initial period of about a day in each exposure was required for insects in diapause to be killed. At 25°C, a concentration of 0.03 mg/l was below the threshold level for mortality of some larvae, long exposures at such very low concentrations proving ineffective.

Diapausing pupae of an outdoor species, Pieris brassicae (L.), and eggs of the univoltine race of Bombyx mori (L.) were highly tolerant of phosphine at 10°C.     

Effects of temperature and humidity on development of four pyralid moth pests of stored products

The development of Plodia interpunctella, Ephestia cautella, E. kuehniella and E. elutella was followed carefully at selected temperatures. Infertility prevented successful breeding of the latter two species at 30°C. The first two species bred successfully at 30°C, failed to complete development at 15°C, and, at 20–30°C, completed development more quickly than the other two. All species took longer to develop and survived less well at 25% r.h. than at 70% r.h., and their pupal stages lasted about twice as long as the egg stages at temperatures allowing development of both. At 25°C, the pupal stage lasted about 9 days in P. interpunctella and E. cautella, 11–12 days in E. kuehniella and about 13 days in E. elutella.

In all species other than E. elutella, eggs held at 25°C became more tolerant of cold when aged over 1 day. Younger eggs of P. interpunctella were less tolerant of cold than any eggs of the other species, and were killed outright by holding for 19 days at 15°C.

Stocks of all four species recently collected from the field generally tolerated low r.h. and low temperature a little better than laboratory stocks, but at higher temperatures developed from egg to adult a little more slowly.     

The termination of diapause in Trogoderma granarium Everts (Coleoptera, Dermestidae)

When subjected to an increase in temperature from 30 to 37°C, both the density-dependent diapause (DDD) and the density-independent diapause (DID) of Trogoderma granarium were terminated in about 10 days. However, under severe crowding, a few DDD larvae took as much as 100 days to pupate at 37°C.

At 30°C, most larvae that had entered diapause in isolated cultures pupated spontaneously during a brief period sometime after they were 3 months old; for most populations of larvae studied such pupation occurred at about 5 months of age. Renewal of food had little influence on the termination of DID. A corresponding burst of pupation, although of only one quarter of individuals, was also recorded for DDD larvae kept crowded in old flour. The most effective termination of DDD, at 30°C, was achieved by isolating the larvae and providing new food together. This treatment brought about a small immediate burst of pupation and also enhanced the subsequent spontaneous pupation so that altogether about 86 per cent of the larvae pupated.

The adaptive value of these reactions in the warehouse environment is discussed. It is suggested that the significance of some of the features of the diapause of T. granarium may be explicable only in relation to its primitive habitat, possibly the nests of insects, in which the diapause must have originated.     

The susceptibility of the immature and adult stages of Sitophilus granarius to phosphine

A concentration of 4 mg/l. of phosphine applied for 8 hr at 25°C permitted a considerable survival of the developmental stages of Sitophilus granarius (L.). Eggs of 3–4 days old were very tolerant and pupae even more so but larvae were very susceptible. A concentration of 1 mg/l. applied for 32 hr completely killed all stages except pupae. The most tolerant eggs were 2–4 days old. At 0·5 mg/l., pupae were again the most tolerant stage and eggs were most tolerant at 1–2 days old. At 0·3 mg/l. or less the majority of eggs were killed during the fifth day of development irrespective of their age at the start of fumigation, but over half were dead by 4 days old at 0·4 mg/l. and about a third survived beyond the sixth day at 0·05 mg/l. It is only at these latter concentrations at these ages that mortality of eggs was clearly related to dosage.

The physiological age range of pupae in samples was more dispersed. Pupae were most tolerant to phosphine at about 32 days old. From 5 to 7 days were needed for complete mortality at concentrations of 0·1–0·05 mg/l.

Adults were very susceptible to phosphine, all being killed by 8 hr at 1 mg/l. which permitted survival of some larvae. Freshly emerged adults were the most tolerant; older adults were very susceptible, at 1 month old being killed by 4 hr at 1 mg/l. and 3 days at 0·01 mg/l. Insects surviving fumigation as larvae or pupae usually laid fewer eggs than controls though their longevity was unaffected. Those surviving fumigation as eggs were not adversely affected. There is a possibility that some individuals surviving, as eggs or pupae, a fumigation causing very high mortality may lay more rapidly than controls.     

Dispersal of three species of grain beetles as a function of thermal acclimation, temperature, and larval size

Three species of stored-product insect pests, Tenebrio molitor, Tribolium confusum, and Trogoderma parabile, were acclimated at three temperatures, 15°, 23°, and 30° or 33°C, and then placed in chambers containing an appropriate culture medium held at testing temperatures ranging from 3°C to 14°C.

Cold-acclimated insects demonstrated greater dispersal at low temperatures than did warm-acclimated insects. Dispersal was greater at higher than at lower test temperatures. In a given species large larvae generally showed greater dispersal than did small larvae.     

The mortality of Rhyzopertha dominica (F.) (Coleoptera: Bostrychidae) and Sitophilus oryzae (L.) (Coleoptera: Curculionidae) at moderate temperatures

A method using conductive heating is described for quickly obtaining and maintaining moderate temperatures in grain while minimising grain moisture loss. A model is developed to analyse insect mortality at a range of conditions on log-time logit and log-time probit scales. Values of LT 50, LT 99, and LT 99.9 are presented for adult and immature stages of Sitophilus oryzae in wheat from 42 to 48°C at 12% m.c. and for immature stages of Rhyzopertha dominica in wheat from 45 to 53°C at 9, 12 and 14% m.c. Data show that the heat tolerance of all life cycle stages for both species are different, with a degree of progressive change relative to each other as temperature changes. For R. dominica, the thermal tolerance of life cycle stages also changes relatively with changes in grain moisture content. In general, all stages survive longer at a given temperature as grain moisture increases and the effect of moisture on survival increases as temperature decreases. Values for LT 99.9 for the most heat tolerant stage of R. dominica at 9% m.c. range from 78.22 to 2.49 h between 45 and 53°C. At 12% m.c. they range from 96.81 to 3.36 h, and at 14% m.c. from 114 to 4.14 h. In general, at 12% m.c. the heat tolerance of S. oryzae is about 5°C less than R. dominica with LT 99.9 ranging from 37.36 to 3.71 h between 42 and 48°C. At this level of mortality, immature stage three is generally the most heat tolerant stage of R. dominica, whereas it is stage two for S. oryzae. The results are discussed in relation to practical applications of heat disinfestation and in terms of a general model of mortality. The results are also considered in the context of other available information on the heat treatment of both species.     

The effect of carbon dioxide gas alone or in combinations on the mortality of Tribolium castaneum (Herbst) and T. confusum du Val (Coleoptera, Tenebrionidae)

Adults of Tribolium castaneum (Herbst) and T. confusum du Val exposed to various mixtures of N₂ or He and O₂ were killed when the O₂ concentration reached to 1·7% or below, whereas the adults exposed to CO₂ : O₂ mixtures were killed mostly due to the deleterious effects of CO₂ itself. At 26·7°C and 38 ± 6%r.h., 95 per cent mortality of T. confusum adults was obtained by an exposure of 271 hr to 45% CO₂ : 55% air mixture; 58 hr to 62% CO₂ : 38% air mixture; and 47·5 hr to 80% CO₂ : 20% air mixture, while for T. castaneum adults 95 per cent mortality required 192, 60, and 44 hr respectively. Data obtained by exposing mature and immature stages of both species to 100% CO₂ suggest that adults were the most susceptible, followed by larvae, eggs and pupae. Generally speaking, inactive stages (egg and pupal) were more tolerant to CO₂ than active stages (larval and adult). Increasing the temperature from 15·6° to 26·7°C resulted in increased insect mortalities; the degree of response varying in different stages. Increasing r.h. decreased the susceptibility of adult insects to 100% CO₂.

Under airtight conditions 200 adult T. castaneum with 8 g of food medium sealed in 1·2 1. glass flasks depleted the O₂ supply from 20·9% to a critical 1·7% level in 7 days, and adults of T. confusum depleted to the 1·6% level in 5 days of airtightness, and both species produced about 20% of carbon dioxide gas.     

Efficacy of the amorphous silica dust, Dryacide, against Tribolium confusum Duv. and Sitophilus granarius (L.) (Coleoptera: Tenebrionidae and Curculionidae)

Amorphous silica dust (Dryacide) was used to treat wheat at concentrations of 0, 250, 500, 750, and 1000 μg silica dust/g wheat. Adult Sitophilus granarius and Tribolium confusum were placed in the grain which was then incubated at 20 or 30°C and 40 or 60% r.h. Mortality counts were taken after 48 and 168 hr. S. granarius was more susceptible to silica dust than T. confusum under the same conditions. At the same temperature, the toxicity of silica dust to both species was more pronounced at 40 than 60% r.h. T. confusum was more tolerant to silica dust at 30 than at 20°C; whereas, S. granarius was more susceptible to it at 30 than 20°C. Silica dust reduced progeny 100% at 40% r.h. in all the concentrations that had been used. Progeny, however, were produced by S. granarius at 30°C and 60% r.h. but with significantly reduced numbers with increasing dosge. Silica dust had no adverse effect on wheat seed germination, wheat flour, and baking quality.     

The tolerance of developmental stages of four stored product moths to phosphine

The tolerance to phosphine of different stages of Ephestia elutella, E. kuehniella, E. cautella and Plodia interpunctella was followed closely at 10–30°C. Eggs of all species were highly tolerant for the first 30–45% of the developmental period, but all other stages were much more susceptible. Maximum exposure periods permitting some survival over a practical range of concentrations were 1 day at 30°C, 2 days at 25°C, 3–4 days at 20°C, and 5–8 days at 15°C. Some eggs of E. elutella survived the very high CT products of 142 mg h/l. in a 2-day exposure at 25°C, and 288 mg h/l. in an 8-day exposure at 15°C. In a 3-day exposure at 25°C, and a 9-day exposure at 15°C, CT products of about 4 and 20 mg h/l. respectively were sufficient for complete control of all species. Eggs of P. interpunctella were susceptible to low temperature and survived exposures to phosphine at 15°C less well than other species. Survivors from fumigated eggs were of normal fertility.Although much less tolerant than eggs, younger pupae, especially those of E. kuehniella, were more tolerant than larvae or older pupae. The fertility of adults of normal weight emerging from surviving pupae was markedly depressed.In most fumigation experiments, differences in tolerance between laboratory and wild stocks were small. However, wild stocks tended to be more tolerant of cold than laboratory stocks, and survived fumigation better in long exposures at lower temperatures.     

Simulation studies on the effects of solar heat on egg-laying, development and survival of Callosobruchus maculatus (F.) and Callosobruchus subinnotatus (Pic) in stored bambara groundnut Vigna subterranea (L.) Verdcourt

The effects of simulated solar heat on oviposition, development and survival of Callosobruchus maculatus (F.) and Callosobruchus subinnotatus (Pic) in stored bambara groundnut, Vigna subterranea (L.) Verdcourt, were evaluated at three high temperatures (40°C, 45°C and 50°C) at a constant, low humidity (30% relative humidity). Exposure to these temperatures for 6 h significantly reduced oviposition in C. maculatus and C. subinnotatus females. Females of both species that were exposed to 50°C laid significantly fewer eggs than those exposed to 40°C; and in the case of C. maculatus, females exposed to 45°C also laid significantly fewer eggs than those exposed to 40°C. The percentage of eggs laid by females of both species that reached adulthood after exposure to 50°C for 2–6 h was significantly lower than the percentage that developed from eggs laid by females that were exposed to 40°C. No adult developed from eggs of C. maculatus exposed for 6 h at 50°C or from eggs of C. subinnotatus exposed for 2 h at this temperature. For both species, no adult progeny subsequently emerged from seeds harbouring first instar larvae when exposed at 50°C for 2, 4 or 6 h. Older larvae of C. maculatus were more tolerant of exposure at 50°C: 26.8, 10.2 and 0.9% of late instar larvae exposed for 2, 4 and 6 h, respectively, developed to the adult stage. In contrast, no adults of C. subinnotatus emerged from seeds harbouring late instar larvae when exposed at 45°C for 6 h nor in seeds exposed to the temperature of 50°C for 4 or 6 h. On average, immature stages of C. subinnotatus were more susceptible to heat treatment than those of C. maculatus.     

Temperature and relative humidity profiles during heat treatment of mills and its efficacy against Tribolium castaneum (Herbst) life stages

Heat treatment involves raising the temperature of a food-processing facility to 50–60°C and maintaining these elevated temperatures for 24–36 h to kill stored-product insects. The pilot feed and flour mills at Kansas State University, Manhattan, KS, were subjected to heat treatment using gas and steam heaters, respectively, during 4–8 August 2001 to characterize temperature and relative humidity profiles and to determine efficacy against developmental stages of the red flour beetle, Tribolium castaneum (Herbst). Temperature and relative humidity profiles in each mill were electronically monitored in 10 different locations. Susceptibility of eggs, young instars, old instars, pupae, and adults of T. castaneum to heat was determined at the same 10 locations within each mill. The number of hours required to reach the target temperature of 50°C, number of hours above 50°C, and maximum temperatures varied between the mills and among mill locations. Relative humidity decreased predictably with an increase in temperature, and was 21% in most locations at 50°C or above. Two separate three-parameter non-linear regression models best described temperature and relative humidity profiles observed in each mill. Despite non-uniform heating at the sample locations, mortality of T. castaneum life stages was 100% in most locations, except in areas where temperatures were <50°C. Old instars and pupae appeared to be relatively heat tolerant when compared with other life stages, especially in the flour mill where lethal temperatures were attained. Possible reasons for the non-uniform heating observed at mill locations and survival of certain T. castaneum developmental stages are discussed.     

Effect of low pressures on the survival of three cocoa pests at 30°C

This study forms part of an effort to eliminate the need for methyl bromide fumigation to control insects in stored commodities, through development of a novel “vacuum-hermetic” technology. The effects of low pressures and exposure times on the mortality of insects in stored cocoa beans were studied at a temperature of 30°C in order to simulate cocoa bean storage conditions in tropical climates. Insects were exposed within test chambers containing the cocoa beans at a moisture content in equilibrium with 55% r.h. and at a constant temperature of 30°C. Three species of insects were used, all being major pests of cocoa beans in producer countries: Ephestia cautella (Walker), Plodia interpunctella (Hübner) and Tribolium castaneum (Herbst). At 50±5 mmHg, the egg stage was the most resistant in all three species, times needed to obtain 99% mortality being 45, 49 and 22 h, respectively. Results show that low-pressure treatment can provide an additional and more effective option to the 5 days fumigation with phosphine used today in the replacement of methyl bromide. The use of low pressures allows the control of insect pests at shorter exposure times without the need for toxic chemicals with their environmental impact.     

Fumigation with carbonyl sulfide: a model for the interaction of concentration, time and temperature

The new fumigant carbonyl sulfide offers an alternative to both methyl bromide and phosphine as a grain fumigant. Separate mathematical models for levels of kill, based on quantitative toxicological studies were developed for adults and eggs of the rice weevil Sitophilus oryzae (L.). These models suggest that fumigation exposure times for carbonyl sulfide will be a compromise between those of methyl bromide (typically 24 h) and phosphine (7–10 d) to achieve a very high kill of all developmental stages. S. oryzae eggs were more difficult to kill with carbonyl sulfide fumigation than the adults. At 30°C, a 25 g m⁻³ fumigation killed 99.9% of adults in less than 1 d, but took 4 d to kill the same percentage of eggs. Models were generated to describe the mortality of adults at 10, 15, 20, 25 and 30°C. From these models it is predicted that fumigation with carbonyl sulfide for 1–2 d at 30 g m⁻³ will kill 99.9% of adults. Furthermore the models illustrate that fumigations with concentrations below 10 g m⁻³ are unlikely to kill all adult S. oryzae. Significant variation was observed in the response of eggs to the fumigant over the temperature range of 10 to 30°C. Models were generated to describe the mortality of eggs at 10, 15, 20, 25 and 30°C. As the temperature was reduced below 25°C, the time taken to achieve an effective fumigation increased. Extrapolating from the models, a 25 g m⁻³ fumigation to control 99.9% of S. oryzae eggs will take 95 h (4 d) at 30°C, 77 h (3.2 d) at 25°C, 120 h (5 d) at 20°C, 174 h (7.5 d) at 15°C and about 290 h (11 d) at 10°C. The role of temperature in the time taken to kill eggs with carbonyl sulfide cannot be ignored. In order to achieve the desired level of kill of all developmental stages, the fumigation rates need to be set according to the most difficult life stage to kill, in this instance, the egg stage.     

Predicting insect density from probe trap catch in farm-stored wheat

Insect populations infesting wheat stored in four bins on two Kansas farms were monitored from early July 1996 through to the middle of January 1997. Estimates of adult insect density based upon the numbers of adult insects caught using probe traps differed from those based upon the number of insects found in grain samples. These differences were a result of differences in numbers of insects found and percentages of traps or grain samples with insects. Traps detected insects 15 to 37 d earlier than grain samples. The depth of traps below the grain surface tended to influence both the total number and species composition of the insects that were caught. Traps inserted with the top just below the grain surface collected an average of 1.9 times more Cryptolestes ferrugineus (Stephens), 1.2 times more Rhyzopertha dominica (F.), 4.1 times more Ahasverus advena (Waltl) and 77.4 times more Typhaea stercorea (L.) than traps inserted with the top 7.6 cm below the grain surface. However, trap depth did not have a significant effect on the number of R. dominica caught and on only 12 to 21% of sampling dates did trap depth have a significant effect on the number of insects of other species that were caught. Grain temperatures in three of the bins averaged 30°C during the first 70 d of storage and then decreased by 0.2°C/d. Grain in the other bin was initially more than 10°C warmer and grain temperature decreased by 0.2°C/d over the full storage period. The numbers of insects captured in traps decreased as grain temperature decreased even though grain samples indicated that insect populations were still growing. Thus, trap catches did not estimate insect population density consistently throughout the storage period. A method was developed in the current paper to adjust for the effect of seasonal changes in temperature on trap catch.     

Effect of high temperatures on larvae of Ephestia elutella (Lepidoptera: Pyralidae) in diapause

Diapausing larvae of Ephestia elutella (Hb.) were exposed at 40, 43 or 45°C and then transferred to 25°C to observe subsequent development and survival. All larvae died after 96 hr at 40°C, 24 hr at 43°C or 16 hr at 45°C. However, about 80% of larvae exposed to 40°C for 8 hr each day for 18 days and about 50% of larvae exposed at 43°C for 8 hr each day for seven days survived. At 45°C all larvae died after two 8-hr or three 6-hr exposures given on successive days, but eight 4-hr exposures failed to kill more than 25% of larvae exposed. When the gap between 6 and 8-hr exposures was increased from 1 day, some larvae survived, indicating that recovery from heat exposure could occur. Adults emerging after high temperature exposure of diapausing larvae proved fertile. In contrast adults from larvae pupating without diapause at 30°C are known to be infertile.

Exposures at high temperature significantly hastened the termination of diapause. In general, the higher the level of mortality in the sample, or the greater the number of exposures at high temperature, the sooner pupation occurred in long days at 25°C.     

Comparative life history studies of four Callosobruchus species infesting cowpeas with special reference to Callosobruchus rhodesianus (Pic) (Coleoptera: Bruchidae)

A comparative study of the ecology of Callosobruchus analis (F.), C. chinensis (L.), C. rhodesianus (Pic) and two strains of C. maculatus (F.) was made at three temperatures (25, 30 and 35°C) and three relative humidities (60, 70 and 80% r.h.). The performances of C. rhodesianus and the strain of C. maculatus from Brazil were also compared at 20°C. The intrinsic rates of increase of each species at each temperature were estimated. The optimum temperature for multiplication of all species was around 30°C. Total oviposition was highest at 30°C for all species, but survival from egg to adult was highest at 25°C for C. rhodesianus and the Malawi strain of C. maculatus compared with 30°C for the others. Development of both strains of C. maculatus was fastest at 35°C, but all other species developed most rapidly at 30°C. Relative humidity in the range 60–80% had no appreciable effect on any of the biological characteristics studied. The performance of C. rhodesianus was better at 20°C than that of the Brazilian strain of C. maculatus, and the results are discussed in relation to the distribution of the two species in Africa.     

Susceptibility of stored-product beetles on wheat and maize treated with thiamethoxam: effects of concentration, exposure interval, and temperature

Sitophilus zeamais Motschulsky, the maize weevil, Oryzaephilus surinamensis (L.), the saw-toothed grain beetle, and Tribolium castaneum (Herbst), the red flour beetle, were exposed for 1, 2, 3, and 6 d at 22°C, 27°C, and 32°C on maize treated with 0, 0.5, 1.0, 2.0, or 4.0 ppm thiamethoxam, a new-generation neonicotenoid insecticide. A second series of tests was conducted on hard winter wheat using S. oryzae (L.), the rice weevil, Rhyzopertha dominica (F.), the lesser grain borer, and T. castaneum. Mortality of all species on both commodities generally increased with insecticide concentration, exposure interval, and temperature, and data were described by linear and non-linear regressions with concentration as the independent variable. Mortality of S. zeamais ranged from 58% to 90% on maize treated with 0.5 ppm thiamethoxam, and approached 95–100% as concentration increased to 4 ppm. Oryzaephilus surinamensis appeared to be slightly less susceptible than S. zeamais; mortality ranged from about 18% to 80% at 5 ppm and there was a more gradual increase in mortality as concentration increased. Mortality of T. castaneum generally did not exceed 40% at any concentration unless the beetles were exposed for 6 d. Mortality of R. dominica and S. oryzae was less than 60% when exposed on treated wheat for 1 and 2 d, but increased to nearly 100% when exposed for 6 d at 27°C and 32°C. Mortality of T. castaneum did not exceed 20% at the 1- and 2-d exposures, and approached 100% only when beetles were exposed for 6 d at 32°C. Few F₁ adults of any species were found in treated maize or in treated wheat but the number of F₁ T. castaneum in untreated maize and untreated wheat was very low compared with the other species. Results show that thiamethoxam would be an effective protectant of stored maize seed and stored wheat seed.     

Physiological properties of Lactobacillus paracasei, L. danicus and L. curvatus strains isolated from Estonian semi-hard cheese

Growth and survival of Lactobacillus paracasei (six strains), L. danicus sp. nov. (four isolates, two strains) and L. curvatus (two strains) from semi-hard Estonian cheeses were comparatively studied in different environmental conditions of relevance for their growth in cheese and survival in gastric environment. Maximum specific growth rates for L. paracasei strains varied between 0.40 and 0.57 h⁻¹, and all strains were tolerant to low water activities, heating at 60 °C for 30 min and pH 3. The newly discovered genetically distinct species L. danicus was characterized by low maximum growth rates (0.26–0.38 h⁻¹) and low temperature optimum (<30 °C). It was acid and heat sensitive and inhibited at salt concentrations from 4% and water activities below 0.93. L. curvatus was characterized by the highest growth rates (0.65–0.70 h⁻¹), tolerance to high NaCl concentrations, but sensitivity to heating, bile salts and low pH. The study showed that genetically different LAB species isolated from cheese could be distinguished by simple cultivation experiments.     

A novel use of modified atmospheres: Storage insect population control

The research described here aimed to establish the feasibility of using modified atmospheres (MA) to protect commodities throughout their storage life by using oxygen (O₂) levels that disrupt the life cycles of the target beetle species. Rather than achieving complete mortality of all stages, the aim was to identify more easily obtainable MAs that would kill the most susceptible stage and prevent population growth. Simulated burner gas and nitrogen (N₂) atmospheres with O₂ contents between 3% and 6%, were tested, along with a N₂-based MA with elevated carbon dioxide (CO₂) (10–20%).

Laboratory tests were carried out on five species of stored-product beetles, Cryptolestes ferrugineus, Oryzaephilus surinamensis, Sitophilus granarius, S. oryzae and Tribolium castaneum. After exposure to the MAs for 28 d an assessment was made of the mortality of adults, the number of adults from progeny produced under the MAs and, for the simulated burner gas, the number of adults from progeny produced in a 28-d period after exposure to the MA. The tests were carried out at 20 and 25 °C with 75% and 85% r.h. at each temperature.

The O₂ content preventing population growth varied with species and temperature. For simulated burner gas or N₂ it was about 4% for O. surinamensis, S. granarius and S. oryzae, and about 3% for C. ferrugineus and T. castaneum at 25 °C. At 20 °C it was about 3% for all species tested. When CO₂ was increased to 10% or 20%, reducing O₂ to 5% was sufficient to eliminate emergence of S. granarius at 20°C, but a few individuals emerged at 25 °C. For C. ferrugineus there was a 95% reduction with 5% O₂ plus 20% CO₂ at 20 °C, but not at 25 °C.     

Temperature and water activity effects on growth and temporal deoxynivalenol production by two Argentinean strains of Fusarium graminearum on irradiated wheat grain

The ability of six strains of Pichia anomala, four strains of Pichia kluyveri and two strains of Hanseniaspora uvarum predominant during coffee processing to produce polygalacturonase (PG), pectin esterase (PE) and pectin lyase (PL) in yeast polygalacturonic acid medium (YPA) and in coffee broth (CB) was studied. For comparison, a reference strain of Kluyveromyces marxianus CCT 3172 isolated from cocoa and reported to produce high amount of PG was included.

Initial screening of PG activity using YPA medium showed that K. marxianus CCT 3172, P. anomala S16 and P. kluyveri S13Y4 had the strongest activity. Enzymatic assays showed that the four yeast species secreted PG, but none of the yeasts investigated was found to produce PE or PL. P. anomala S16 and P. kluyveri S13Y4 were found to produce higher amounts of PG when grown in CB than in YPA. When K. marxianus CCT 3172, P. anomala S16 and P. kluyveri S13Y4 were grown in YPA broth adjusted to pH of 3.0–8.0 and incubated at temperatures of 15–40 °C, the three yeast species secreted the highest amount of PG at pH 6.0 and at 30 °C. For PG secreted by K. marxianus CCT 3172 and P. anomala S16, the optimum pH and temperature for the enzymatic activity were 5.5 and 40 °C, respectively. On the other hand, PG produced by P. kluyveri S13Y4 showed the highest activity at pH 5.0 and 50 °C.

Significant differences in the extracellular activity of PG were found between the yeasts species as well as between strains within same species. High amounts of PG were produced by two strains of P. anomala and P. kluyveri. It is therefore likely that strains of those two species may be involved in the degradation of pectin during coffee fermentation.