Effect of low pressure on the survival of Trogoderma granarium Everts, Lasioderma serricorne (F.) and Oryzaephilus surinamensis (L.) at 30 °C

The effects of low pressure (50 mm Hg) and various exposure times were studied on the mortality of three important storage pest insects at 30 °C as 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. Insects were exposed within test chambers containing cocoa beans with a moisture content in equilibrium with 55% relative humidity and at a constant temperature of 30 °C. Three insect species were used: Trogoderma granarium, Lasioderma serricorne and Oryzaephilus surinamensis. At 50±5 mm Hg and 30 °C, the egg was the most resistant stage in all three species at the LT₉₉ level, times needed to obtain 99% mortality being 46, 91 and 32 h, respectively. Adults of T. granarium and L. serricorne, and pupae of O. surinamensis were most susceptible. These results widen the database established to kill insects at low pressure within the framework of practical exposure times and commercially available equipment, in order to provide an effective alternative to fumigation.     

Studies on insect infestation in chocolates

The ability of stored-product pests including the cigarette beetle, Lasioderma serricorne, the sawtoothed grain beetle, Oryzaephilus surinamensis, the rust-red flour beetle, Tribolium castaneum, and the almond moth, Ephestia cautella, to infest chocolates under packaged and unpackaged conditions was investigated in the laboratory at 25±1 °C and 65±5% r.h. Four types of chocolates were investigated: milk, nut, dried fruit and nut, and wafer chocolates. Adults (beetles only, 20 per replicate) or eggs (30 per replicate) were released on unpackaged and packaged chocolates and infestation levels (number of living adults and larvae) were determined 45 days later. When adult beetles were released on unpackaged chocolates, the degree of infestation varied depending on the species and the type of chocolate. The highest infestation observed in unpackaged chocolate was that of O. surinamensis in wafer chocolate (mean 138.4). When eggs were released on unpackaged chocolates, the most numerous species was E. cautella in dried fruit and nut chocolate (mean population=180.8). With packaged chocolates exposed to adults or eggs, insect infestation was nil or negligible (mean population <6.0). Although infestation levels were low, infestations were found in 50% of treatments over all. Damage to the packaging material along the folds or edges was observed in infested chocolates. The study has shown that milk, nut, dried fruit and nut, and wafer chocolates can support insect infestation and therefore, insect-proof packing of the chocolates and storage under hygienic conditions are important to avoid customers’ complaints.     

Initial and delayed mortality of late-instar larvae, pupae, and adults of Tribolium castaneum and Tribolium confusum (Coleoptera: Tenebrionidae) exposed at variable temperatures and time intervals

Late-instar larvae, pupae, and adults of Tribolium castaneum (Herbst), the red flour beetle, or Tribolium confusum (DuVal), the confused flour beetle, were exposed for variable durations at 36-54 °C. Beetles were placed in laboratory ovens set at a baseline of 27 °C, the temperature was increased by 0.1 °C per minute until the target temperature was achieved, and beetles were then held for specified exposure durations. There was no mortality after initial exposure or after a 1-week holding period of any life stage of T. castaneum or T. confusum exposed for 32 h to 36, 39, or 42 °C. At 45 °C, there was no initial mortality of either species exposed for different time intervals except for those exposed for 28 h. However, there was a significant increase in mortality after the 1-week holding period of those beetles exposed initially for at least 16 h to 45 °C. There was a sharp increase in mortality after the initial exposures of 4 h at 48 °C; mortality of T. confusum larvae was 90.0±5.7% but was only 10.0±10.0% for larvae of T. castaneum, and no pupae of either species were dead. All life stages of both species were killed after the initial exposure of 12 h, and 1-week mortality of beetles exposed for 4 and 8 h was generally greater than initial mortality. At 51 and 54 °C, 2- and 1-h exposures, respectively, killed all life stages of each species. Mortality in conditions of gradual temperature increase was less than previous studies with sudden temperature increases.     

Effect of “soft-electron” (low-energy electron) treatment on three stored-product insect pests

Developmental stages of three stored-product insect pests viz. Tribolium castaneum (Herbst), Plodia interpunctella (Hübner) and Callosobruchus chinensis (L.) were exposed to “soft electrons”, low-energy electrons. Soft electrons at an acceleration voltage of 170 kV effectively inactivated eggs, larvae and pupae of T. castaneum and P. interpunctella. The adults of T. castaneum and P. interpunctella were inactivated by treatment for 10 min (4.8 kGy) and 15 min (7.2 kGy), respectively. Soft electrons at 170 kV also inactivated the eggs of C. chinensis effectively. The adults of C. chinensis survived a 15 min exposure (7.2 kGy), but were inactivated having lost the ability to walk after a 5 min exposure (2.4 kGy). Soft electrons at 170 kV could not completely inactivate the larvae of C. chinensis inside beans, because the electrons with low penetration did not reach the larvae due to the shield of beans. The movement of T. castaneum larvae exposed to soft electrons was sluggish for several days, and then the larvae died with their hindguts emerged from their posterior ends 17 days after the treatment. DNA comet assay of cells of P. interpunctella larvae indicated that soft electrons at 170 kV damaged DNA in the larvae.     

Impact of resistance on the efficacy of binary combinations of spinosad, chlorpyrifos-methyl and s-methoprene against five stored-grain beetles

Laboratory experiments were conducted to determine the efficacy of spinosad (a biopesticide), chlorpyrifos-methyl (an organophosphorus compound (OP)) and s-methoprene (a juvenile hormone analogue) applied alone and in binary combinations against five stored-grain beetles in wheat. There were three strains of Rhyzopertha dominica, and one strain each of Sitophilus oryzae, Tribolium castaneum, Oryzaephilus surinamensis and Cryptolestes ferrugineus. These strains were chosen to represent a range of possible resistant genotypes, exhibiting resistance to organophosphates, pyrethroids or methoprene. Treatments were applied at rates that are registered or likely to be registered in Australia. Adults were exposed to freshly treated wheat for 2 weeks, and the effects of treatments on mortality and reproduction were determined. No single protectant or protectant combination controlled all insect strains, based on the criterion of >99% reduction in the number of live F₁ adults relative to the control. The most effective combinations were spinosad at 1 mg kg⁻¹+chlorpyrifos-methyl at 10 mg kg⁻¹ which controlled all strains except for OP-resistant O. surinamensis, and chlorpyrifos-methyl at 10 mg kg⁻¹+s-methoprene at 0.6 mg kg⁻¹ which controlled all strains except for methoprene-resistant R. dominica. The results of this study demonstrate the difficulty in Australia, and potentially other countries which use protectants, of finding protectant treatments to control a broad range of pest species in the face of resistance development.     

Heat treatment for disinfestation of empty grain storage bins

An alternative to fumigants and insecticides for controlling stored-product insects in empty grain storage bins prior to filling is heat treatment, in which the temperature is quickly raised to a minimum of 50 °C and held there for 2-4 h. Effectiveness of heat treatment on empty grain storage bins was evaluated for five commercial propane and electric heat-treatment systems by measuring air temperature and associated mortality of Tribolium castaneum (Herbst), the red flour beetle, Sitophilus oryzae (L.), the rice weevil, and Rhyzopertha dominica (F.), the lesser grain borer, exposed for different time intervals. Eleven locations, six above and five below the drying floor, were monitored for air temperature and associated mortality of the three insect species, using arenas initially stocked with live adult insects. Data were analyzed separately for each heating system, with floor location and time interval as main effects for insect mortality. A high-output propane heater (29 kW) produced 100% mortality in 2 h for the three insect species at all test locations. An electric duct-heater system (18 kW) also produced 100% mortality at all test locations after 40 h when aided by a complicated interior heat-distribution system. The other three systems produced less than 100% mortality.     

Effectiveness of short exposures of propylene oxide alone and in combination with low pressure or carbon dioxide against Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae)

Toxicity of the fumigant propylene oxide (PPO) alone and in combination with low pressure (100 mm Hg) or 92% CO₂ to all life stages of Tribolium castaneum using short exposure times (4 and 8 h) at 30°C was studied. Results indicated that PPO was moderately toxic with Ct products ranging from 120 to 608 mg h/l required to obtain complete mortality of the different life stages. A marked difference in susceptibility between life stages was recorded. Eggs were the most sensitive with a LD₉₉ value of 30.1 mg/l for 4 h, whereas pupae were the most tolerant with a LD₉₉ value of 146.5 mg/l. It was shown that an increase in exposure time from 4 to 8 h resulted in 23%, 42%, 48% and 47% reductions of LD₉₉ values for eggs, larvae, pupae and adults, respectively.There was no or very limited mortality of all stages except the egg (53% to 62%), when exposed to either 100 mm Hg or 92% CO₂ for 4 h. However, when 100 mm Hg or 92% CO₂ were combined with PPO, the LD₅₀ and LD₉₉ values for PPO in all stages except the egg were significantly reduced. Combinations of PPO with 100 mm Hg or 92% CO₂ produced equal reductions in the LD₉₉ value from 146.5 to about 22 mg/l for the most tolerant pupal stage. Both combinations also produced significant reductions in the LD₉₉ values for larvae and adults (6.3- to 6.6-fold) compared with those exposed to PPO alone. These results indicated that 100 mm Hg and 92% CO₂ each had a synergistic effect on the toxicity of PPO to T. castaneum. The combination of PPO with vacuum or CO₂ can thus provide a potential alternative to methyl bromide.     

Laboratory assessment of insecticidal effectiveness of natural zeolite and diatomaceous earth formulations against three stored-product beetle pests

The insecticidal effectiveness of two natural zeolite formulations (Minazel plus and Minazel), applied to wheat at selected rates of 0.25, 0.50 and 0.75 g/kg, and a diatomaceous earth formulation (DE) (Protect-It™), applied at the recommended rates of 0.15 g/kg for Sitophilus oryzae, 0.20 g/kg for Rhyzopertha dominica and 0.30 g/kg for Tribolium castaneum, were tested under laboratory conditions (24 ± 1 °C temperature and 45 ± 5% relative humidity). The highest adult mortality was observed after the longest exposure period of 21 days and 7 days of recovery, when all three zeolite dosage rates and the recommended DE dosage caused 97-100% mortality of S. oryzae and 94-100% of T. castaneum. On the other hand, 100% mortality was not achieved in any test variant involving R. dominica; the highest (about 92%) was detected for DE, while 52% and 79% mortality was achieved with the zeolites at the highest rate of 0.75 g/kg. Progeny reduction by >90% was achieved after 21 days of contact of all three beetle pests with DE-treated wheat, while the same level of reduction was achieved for S. oryzae and T. castaenum only after contact with the highest rate of the zeolite product, Minazel. Thus the two zeolite formulations are comparable to diatomaceous earth in controlling adult S. oryzae, R. dominica and T. castaneum, but only the Minazel formulation could effectively protect wheat from attack by S. oryzae or T. castaneum, and only with a higher rate of application than for the DE formulation.     

Impact of high temperatures on efficacy of cyfluthrin and hydroprene applied to concrete to control Tribolium castaneum (Herbst)

In laboratory trials, concrete was treated with cyfluthrin wettable powder (WP) at 40 mg active ingredient [AI] cyfluthrin WP/m², then heated for 4, 8, or 16 h at either 45°C or 55°C, or treated but not heated (seven treatment combinations). Bioassays were conducted by exposing adult Tribolium castaneum (Herbst) for 0.5, 1, and 2 h. Survival of T. castaneum was generally greater on unheated concrete compared with the heating treatments, and survival appeared to decrease as heating time increased at both 45°C and 55°C. In a second laboratory trial, concrete was treated with hydroprene (Gentrol) at the label rate of 1.9×10⁻³ mg [AI]/cm², and bioassayed by exposing late-instar T. castaneum larvae on the treated surface. There were significant differences between untreated controls and the heat treatment regimes (P<0.05) with respect to the percentage of live emerged adults, the percentage of those adults with deformities, and the percentage of dead adults, but heating did not reduce efficacy of hydroprene. In a field trial, concrete was treated with cyfluthrin at 2 mg [AI] cyfluthrin WP/m², and placed in a flour mill undergoing an experimental heat treatment and in an unheated office. Treated concrete was bioassayed by continually exposing adult T. castaneum for 0.5-120 h. The effect of heating time on insect mortality was not significant (P?0.05). Except for T. castaneum exposed for 0.5 h, the percentage of beetle survival on unheated concrete was greater (P<0.05) than survival on concrete that had been heated in the mill, indicating a possible beneficial effect on cyfluthrin toxicity due to heating. Results of these studies show that short-term exposures to high temperatures may have no appreciable effect on efficacy of either cyfluthrin WP or hydroprene, and combination treatments of heat plus either of these insecticides may be effective alternatives to methyl bromide for disinfesting milling facilities.     

Feeding bioassay for stored-product insect pests using an encapsulated food source

A procedure that was developed to encapsulate liquid and semiliquid diets was used to encapsulate dry diet for use in a feeding bioassay for beetles that are pests of stored products. Vacuum was used to form Parafilm^® into numerous 6 mm diameter wells. The wells were filled with clean sand (control) or ground dry dog food (test), and the Parafilm^® sealed to produce individual pellets. A single pellet was then placed in the center of a 9 cm diameter Petri dish and feeding activity of groups of ten adults of Sitophilus oryzae (L.), the rice weevil; Oryzaephilus surinamensis (L.), the sawtoothed grain beetle; and Tribolium castaneum (Herbst), the red flour beetle, was tested. The number of insects on the encapsulated pellet and the amount of food or sand scattered were checked hourly for the first 8 h of the study and after 24 h. Few insects were observed on pellets containing sand and little or no sand was observed scattered outside of the pellet, so the presence of dog food in the pellet was needed for insects to feed on the pellet. In tests with encapsulated dog food, the amount of food scattered provided a better quantitative measure of feeding than the number of insects on the pellet. Insects that were starved for 48 h caused greater amounts of food scatter than insects starved for 24 h prior to the test. In direct comparisons among all three species, T. castaneum responded the most slowly and the bioassay may be improved by increasing the amount of time starved. Sitophilus oryzae responded very quickly and the entire pellet was essentially consumed within the first 4-5 h of the study. The encapsulated diets provide a promising method to evaluate feeding behavior of stored-product insect pests.     

Enhanced fumigant toxicity of allyl acetate to stored-product beetles in the presence of carbon dioxide

Toxicity of allyl acetate (5-25 mg/l doses) to mixed-age cultures of stored-product beetles including Cryptolestes ferrugineus, Lasioderma serricorne, Oryzaephilus surinamensis, Rhyzopertha dominica, Sitophilus oryzae and Tribolium castaneum in the presence of carbon dioxide (CO₂) (10% and 20%) with a 48-h exposure period was studied in the laboratory at 27±2 °C. Depending on dosage and the insect species, the fumigant toxicity of allyl acetate was enhanced by CO₂. At most of the allyl acetate+CO₂ combinations, increased mortality was observed in C. ferrugineus, O. surinamensis and S. oryzae. In L. serricorne and T. castaneum, which are tolerant to allyl acetate, higher mortality due to CO₂ was achieved at selected dose combinations only (e.g. 15 mg/l allyl acetate+20% CO₂). Significant increase in mortality of R. dominica (the most susceptible species to allyl acetate) exposed to allyl acetate in the presence of CO₂ was not evident except at the lower dose of 5 mg/l allyl acetate+CO₂ that caused 31.7% mortality. The mortality data show that CO₂ could be used as an adjuvant for allyl acetate.     

Insect population dynamics in commercial grain elevators

Data were collected in 1998-2002 from wheat stored in commercial grain elevators in south-central Kansas. Bins at these elevators had concrete walls and were typically 6-9 m in diameter and 30-35 m tall. A vacuum-probe sampler was used to collect grain samples in the top 12 m of the wheat in each bin. The primary insect species found in the wheat samples were: Cryptolestes ferrugineus, Rhyzopertha dominica, and Tribolium castaneum. In the top 3.7 m of grain, R. dominica, C. ferrugineus, T. castaneum and Sitophilus oryzae made up 44, 36, 19 and 1% of the insects found in the samples, respectively. From 3.8 to 12.2 m, R. dominica, C. ferrugineus, T. castaneum and S. oryzae were present at 84, 8, 8, and 1%, respectively. The most prevalent species also changed over time. In June, the start of wheat harvesting and storage in Kansas, insect density was low in the bins. At this time, C. ferrugineus was the most common insect, and it was found mostly in the top grain sample (0-1.2 m). In September through November, C. ferrugineus and R. dominica were at similar densities; however, from February to March, R. dominica was more common.Generally, insect density was greatest at the top and decreased with grain depth. Very few insects were found in samples collected from greater than 12 m (most of the bins contained grain to depths of 24-36 m). Insect density for all species increased rapidly from June through October. During this period less than 20% of the bins had economically significant insect densities (>2 insects/kg). From October until February, the average insect density remained fairly constant but it was greatly reduced in April, May, and June. Bins that had insect densities >2 insects/kg tended to be located adjacent to other heavily infested bins.     

Survival and reproduction of Tribolium castaneum (Herbst), Rhyzopertha dominica (F.) and Sitophilus oryzae (L.) following periods of starvation

This study determined the starvation tolerance of Tribolium castaneum (Herbst), Rhyzopertha dominica (F.) and Sitophilus oryzae (L.) in terms of both adult survival and reproduction, the impact of starvation on reproduction not having been studied before. Experiments were conducted at 30 °C and 55% or 70% r.h. using a laboratory strain and a field strain of each species. The number of progeny was a better indicator of the impact of starvation on a species than adult survival. Tribolium castaneum was the most tolerant species, requiring up to 35 d starvation before no progeny were produced. Rhyzopertha dominica and S. oryzae required up to 8 d starvation before no progeny were produced. The results suggest that hygiene will have a greater impact on populations of S. oryzae and R. dominica than T. castaneum.     

Effect of Pisum sativum fractions on the mortality and progeny production of nine stored-grain beetles

Yellow field pea (Pisum sativum L.) fractions that were mainly protein (50%), fibre (90%) or starch (85%) were obtained from a commercial pea mill and mixed with wheat kernels or wheat flour. Based on the mortality and the number of offspring produced, protein-rich pea flour was more toxic than fibre, which was more toxic than starch. For the protein-rich pea flour mixed with wheat kernels, the most sensitive insects were Sitophilus oryzae (L.), Sitophilus zeamais Motschulsky and Sitophilus granarius (L.), followed by Cryptolestes ferrugineus (Stephens) which was more sensitive than Tribolium castaneum (Herbst) and Rhyzopertha dominica (F.). For the protein-rich pea flour mixed with wheat flour, Cryptolestes pusillus (Schönherr) was most sensitive, followed by C. turcicus (Grouvelle) and T. confusum (Jacquelin du Val), with T. castaneum being the most resistant. Although protein-rich pea flour did not kill adults to a great extent when mixed with flour, it reduced offspring production significantly. Again C. pusillus was the most sensitive, followed by T. confusum, with T. castaneum offspring being the most resistant. The insecticidal activity of pea fractions decreased after treated wheat kernels were held at 30 °C, 70% r.h. for 8 months. The potential of using pea fractions to control stored-product insects is discussed.     

Behavior of Salmonella during fermentation,drying and storage of cocoa beans

Due to cocoa being considered a possible source of Salmonella contamination in chocolate, the behavior of Salmonella during some cocoa pre-processing stages (fermentation, drying and storage) was investigated. The fermentation process was carried out on a pilot scale (2 kg beans/box) for 7 days. Every day a fermentation box was inoculated with a Salmonella pool (ca. 4 log MPN/g). The results showed that Salmonella did not affect (P > 0.05) the growth of the main microorganism groups involved in cocoa fermentation. On the other hand, the pathogen was influenced (P < 0.05) by yeast, acetic acid bacteria and pH. In spite of Salmonella showing counts ≤ 1 log MPN/g in the first days, at the end of fermentation it grew in all samples, reaching counts as high as 7.49 log MPN/g. For drying and storage, cocoa beans were inoculated during the fermentation (experiment A) or during the drying (experiment B). In these stages the decline of the water activity affected the pathogen behavior. In experiment A during the drying, Salmonella count increased in most of the samples. In experiment B either a slight growth or no growth in the samples inoculated up to 48 h was observed, whereas the other samples showed reductions from the initial count. After 30 days of storage at room temperature, the water activity decreased to 0.68, and reductions of Salmonella ranged from 0.93 to 2.52 log MPN/g. Despite the reductions observed during the storage, the pathogen was detected even after 120 days. Therefore, the results showed that Salmonella growth or survival depends on when the contamination occurs.     

Toxicity of ethyl formate to adult Sitophilus oryzae (L.), Tribolium castaneum (herbst) and Rhyzopertha dominica (F.)

Fumigations were conducted using a continuous flow-through laboratory process to maintain constant concentrations of ethyl formate and low levels (<0.8%) of respiratory carbon dioxide. The procedure minimised the effects of sorption by exposing test insects without media and minimised the effect of carbon dioxide by use of continuous flow. The concentration×time (Ct) products of ethyl formate for adult Sitophilus oryzae, Tribolium castaneum and Rhyzopertha dominica at 25 °C and 70% relative humidity for the 6 h exposure were, respectively: (1) LD₅₀ 107.8, 108.8 and 72.8 mg h L⁻¹ and (2) LD_99.5 207.4, 167.1 and 122.2 mg h L⁻¹. Endpoint mortality was reached within 24 h of initial exposure.     

Effectiveness of products from four locally grown plants for the management of Acanthoscelides obtectus (Say) and Zabrotes subfasciatus (Boheman) (both Coleoptera: Bruchidae) in stored beans under laboratory and farm conditions in Northern Tanzania

The effectiveness of whole or powdered leaves (botanicals) from four locally grown plant species applied at a rate of 1.5 kg per 100 kg beans (Phaseolus vulgaris) against Acanthoscelides obtectus and Zabrotes subfasciatus was compared under laboratory and farm conditions. In the laboratory, Chenopodium ambrosioides, applied as powder or as whole leaves, was the most effective, with 100% mortality of adult insects in less than three days and no progeny. Less C. ambrosioides (about 200 g per 100 kg beans) still resulted in 100% mortality within 24 h. Tagetes minuta applied as powder also increased mortality and reduced oviposition and progeny production significantly. The other treatments - T. minuta applied as leaves, and Azadirachta indica or Cupressus lusitanica applied as powder or as whole leaves - had no significant effects upon mortalities, oviposition rate, or progeny production compared with control treatments. When the rate of application was increased to about 8.3 kg per 100 kg beans, there was a slight increase in mortality using T. minuta and A. indica, but not with C. lusitanica. An additional trial with C. ambrosioides from different collections and with plants at different stages of development revealed considerable variations in the efficacy of the treatment.In the on-farm trials, A. indica-seed powder was the most effective treatment, followed respectively by leaf powders of C. ambrosioides, C. lusitanica and T. minuta. All treatments were significantly more effective than the control in reducing the numbers of live insects; they also reduced numbers of damaged beans and maintained germination rates after 5 months of storage. The results of evaluations of the treatments made by farmers just after the trials and five years later are reported.     

Moisture content gradient and ventilation in stored wheat affect movement and distribution of Oryzaephilus surinamensis and have implications for pest monitoring

This study was carried out primarily to ascertain whether the movement of Oryzaephilus surinamensis and Sitophilus granarius from low to high humidity zones occurs in bins of wheat and whether aeration of the grain (10 m³/h/t) affects this movement. The second aim was to ascertain the best placement of insect detection traps under the different conditions. Insects were introduced into the lower half of the grain in the bins and their movement was monitored using traps placed at various depths in the grain. Sitophilus granarius did not move through the grain into the top layer regardless of the moisture content, temperature or aeration status of the grain. More O. surinamensis were caught in unventilated bins than in ventilated bins. More insects were caught in the ventilated bins containing layers of both dry and wet grain than in the bins containing only dry grain. The spatio-temporal distribution of O. surinamensis varied significantly. The depth at which insects were trapped varied between treatments: in ventilated dry grain, most insects were trapped at the surface; in ventilated wet and dry grain, most insects were trapped at 10 cm and 0.75 m; in unventilated wet and dry grain, the vast majority of the insects were trapped at 0.75 m. Very few insects were trapped at 1.75 m regardless of the treatment. The proportions of the initial population of O. surinamensis which were recaptured in the top layer of grain varied between treatments. Most were recaptured in the unventilated bins containing wet and dry grain followed by ventilated bins containing wet and dry grain. The smallest proportion of the population was recaptured in the ventilated bins containing only dry grain. Immediate practical implications for pest monitoring based on physical control measures in use are discussed.     

Thermal inactivation of Salmonella spp. during conching

Although chocolate is a microbiologically stable product it has been described as a vehicle for Salmonella spp. Because of the low water activity (a_w) and the high fat content of chocolate Salmonella spp. shows an increased heat resistance, even during the thermal process of chocolate making. The aim of this study was to evaluate the thermal inactivation of Salmonella spp. during conching in various masses of chocolate and cocoa butter at different temperatures (50-90 °C). The effect of thermal treatment on Salmonella spp. was determined with the MPN (Most-Probable-Number) method. Results of thermal treatment showed approximate D-values for cocoa butter from D_50°C = 245 min to D_60°C = 306 min, for cocoa liquor from D_50°C = 999 min to D_90°C = 26 min and for dark chocolate of D_50°C = 1574 min. z-values were found to be z = 20 °C in cocoa liquor and z = 14 °C in dark chocolate. This study demonstrates that the conching process alone does not ensure the inactivation of Salmonella spp. in different chocolate masses and that an additional decontamination step at the beginning of the process as well as an HACCP concept is necessary during chocolate production to guarantee the absence of Salmonella spp. in chocolates and related products.     

The effect of a new chitin synthesis inhibitor, novaluron, on various developmental stages of Tribolium castaneum (Herbst)

Novaluron, a novel chitin synthesis inhibitor (CSI), was tested against the various developmental stages of Tribolium castaneum (Herbst), at concentrations ranging from 0.1 to 100 ppm. It did not kill T. castaneum adults at these concentrations, but at 1.0 ppm it caused total mortality of third-instar larvae. Novaluron did not affect the number of eggs laid by T. castaneum adults that were exposed to treated wheat flour, but it totally inhibited their hatching after the third day of exposure at a concentration of 1 ppm. On the first day after infestation novaluron did not totally prevent hatching, even at the highest tested concentration of 100 ppm. The time needed to restore egg hatchability after adults were transferred to untreated flour depended on the concentration of novaluron used in the treatment. When the adults of T. castaneum were exposed to novaluron-treated whole wheat grains (at 1 ppm), similar effects to those of the treated flour at the same concentration were observed: egg hatching was drastically reduced. The effect of uptake via contact of adults with novaluron-treated surfaces was examined by exposing them to a mixture of untreated flour and 10% novaluron-treated sand at a concentration of 10 ppm, and only 1% of the eggs hatched after 18 days of exposure. When adults were removed from treated surfaces of Petri dishes to untreated flour, the hatching rate was again reduced by 100%. It was concluded that the contact penetration of novaluron into T. castaneum adults prevents the hatching of eggs subsequently laid.