Trap catches of stored-product insects before and after heat treatment in a pilot feed mill

The pilot feed mill at Kansas State University was heated to temperatures of 50°C for 28–35 h during August 4–6, 1999 using natural gas heaters to kill stored-product insects. A three-parameter nonlinear regression model satisfactorily described temperature profiles on each of the four mill floors and was useful in showing differences among floors in the number of hours taken to reach 50°C and hours above 50°C. Pitfall traps with food and pheromone lures and sticky traps with pheromone lures were used to sample adults of beetles and moths, respectively, between July 8 and December 1, 1999 to evaluate heat treatment effectiveness. A total of 32 insect species representing 26 families in seven orders were captured in traps. Immediately after heat treatment, there was 95% reduction in total beetle captures in pitfall traps and 99% reduction in moth captures in sticky traps. Trap captures of the almond moth, Cadra cautella (Walker) and cigarette beetle, Lasioderma serricorne (L.) were significantly reduced and remained low after heat treatment. However, trap captures of the flat grain beetle, Cryptolestes pusillus (Schöenherr), Indianmeal moth, Plodia interpunctella (Hübner), and red flour beetle, Tribolium castaneum (Herbst) increased gradually after heat treatment, especially on the 1st and 4th floors. Our results indicated that traps are valuable tools for gauging the degree and duration of insect suppression obtained by heat treatment. In addition to trapping, visual inspection of the mill areas and absolute sampling of ingredients, products and spillage should be undertaken, so that areas of incipient insect reinfestation can be identified and potential problems rectified or averted.     

Species distribution,antibiotic resistance and virulence traits in enterococci from meat in Tunisia

Antimicrobial resistance and the mechanisms implicated were studied in 119 enterococci from 105 meat samples from Tunisian markets. Almost 24.5% of recovered enterococci showed resistance against four or more antimicrobial agents and these isolates were identified to the species level. Enterococcus faecalis was the most prevalent species (41%). High percentages of erythromycin and tetracycline resistances were found among our isolates, and lower percentages were identified to aminoglycosides, ciprofloxacin and chloramphenicol. All tetracycline-resistant isolates carried the tet(M) and/or tet(L) genes. The erm(B) gene was detected in 78.5% of erythromycin-resistant isolates, ant(6)-Ia gene in 58.8% of streptomycin-resistant isolates, and cat(A) gene in one chloramphenicol-resistant isolate. Forty-eight isolates carried the gelE gene and exhibited gelatinase activity. The hyl and esp genes were detected in one and three Enterococcus faecium isolates, respectively. Streptomycin-resistant isolates showed a high genetic diversity by PFGE and MLST. Meat might play a role in the spread through the food chain of enterococci with these virulence and resistance characteristics to humans.     

Fumigant toxicity of essential oil from Artemisia sieberi Besser against three stored-product insects

Artemisia sieberi is a widely distributed plant in Iran. Because some species of Artemisia are insecticidal, experiments were conducted to investigate fumigant toxicity of the essential oil. Dry ground leaves were subjected to hydrodistillation using a modified Clevenger-type apparatus and the resulting oil contained camphor (54.7%), camphene (11.7%), 1,8-cineol (9.9%), β-thujone (5.6%) and α- pinene (2.5%).The mortality of 7 days old adults of Callosobruchus maculatus, Sitophilus oryzae, and Tribolium castaneum increased with concentration from 37 to 926 μL/L and with exposure time from 3 to 24 h. A concentration of 37 μL/L and an exposure time of 24 h was sufficient to obtain 100% kill of the insects. Callosobruchus maculatus was significantly more susceptible than S. oryzae and T. castaneum; a second more detailed bioassay gave estimates for the LC₅₀ of C. maculatus as 1.45 μL/L, S. oryzae 3.86 μL/L and T. castaneum 16.76 μL/L. These results suggested that A. sieberi oil may have potential as a control agent against C. maculatus, S. oryzae and T. castaneum.     

Detection of phosphine resistance in major stored-product insects in Greece and evaluation of a field resistance test kit

The use of phosphine has been effective against a wide range of stored-product pests in different types of commodities and facilities. However, its continuous and improper use has led to resistance development in -several major insect species. Although phosphine resistance has been reported from many countries across the globe, reports from Europe have been very limited. In the present study, we determined phosphine resistance in insect populations that had been collected from a range of storages across Greece, using two different diagnostic protocols. Apart from the traditional Food and Agriculture Organization (FAO) protocol, a field test kit (known as the Detia Degesch Tolerance Test Kit, DDTTPK) was utilized, for “same day” determination of the resistance status of field collected insects. In total, 53 populations belonging to Rhyzopertha dominica, Sitophilus oryzae, Sitophilus granarius, Cryptolestes ferrugineus, Tribolium confusum, Tribolium castaneum and Oryzaephilus surinamensis were tested. For the majority of the species and populations tested, both FAO and DDPTTK provided similar results, for the susceptibility to phosphine and thus, the quick test could be used with success for an initial same day screening of phosphine resistance. Among the tested species, the populations recorded with the most frequent survival at the FAO testing dose of phosphine was that of R. dominica. The dissimilar evaluation and characterization of resistance to phosphine between diagnostic protocols is particularly important, as it poses risks in the over or underestimation of the resistance status of a given population. Our data indicate that the DDPTTK could be used to determine resistance to phosphine in the field, before the initiation of fumigations to disinfest stored commodities.     

Insecticidal efficacy of a binary combination of cyphenothrin and prallethrin,applied as surface treatment against four major stored-product insects

In the present study, we investigated the insecticidal efficacy of a commercial binary combination of two pyrethroids, cyphenothrin and prallethrin, as structural treatment against adults of Sitophilus oryzae (L.), Oryzaephilus surinamensis (L.), Tribolium confusum Jacquelin du Val and Prostephanus truncatus (Horn) in laboratory bioassays. In a first series of bioassays, the insecticide was applied at its label dose on concrete and metal surfaces, whereas mortality and knockdown levels were recorded after 1, 3, 7 and 14 d of exposure. In a second series of bioassays, the residual efficacy of the insecticide was evaluated on concrete for a period of four months, stored either in continuous darkness or exposed to light. In these bioassays, mortality and knockdown of the exposed insects was measured after 3, 7 and 14 d of exposure. Finally, in a third series of bioassays, we investigated the immediate and delayed effect of the tested formulation on adults of the four insect species after exposure for 1, 3 and 7 d. Based on our results, the application of the tested insecticide as surface treatment can provide efficient control against adults of all four species tested. The surface type did not significantly affect the insecticidal efficacy of the tested formulation, at least for the surfaces tested. Moreover, exposure to light negatively affected the residual activity of the insecticide, as for all insect species tested, mortality was significantly lower on dishes exposed to light than on the ones stored in the dark for storage intervals equal or longer than two months. Finally, reduced delayed mortality was observed for all exposure intervals, as knocked down individuals remained, in most cases, knocked down even 7 d after their removal from the treated dishes.     

Temporal changes in stored-product insect populations associated with boot,pit, and load-out areas of grain elevators and feed mills

Commercial grain elevator and feed mill facilities can quickly become infested with stored-product insect pests, compromising the protection of the stored raw and processed cereal products. Elevators and feed mills were sampled monthly for adult stored-product insects in grain residues from the boot-pit areas and in bulk load-out samples from 2009 to 2010. The grain samples collected from the boot, pit, and load-out areas consisted of corn, soybeans, or a mixture of the two grains. Low insect densities were found in the boot-pit area during the cool winter months. Insect numbers increased in the spring and peaked during the warm summer months, prior to declining in the fall following a pattern of higher insect densities during higher ambient temperatures. The rice weevil, Sitophilus oryzae (L.), was the most prevalent species collected in all of the sampled grain residues, representing 69.2 and 35.8% of total insects collected in feed mills and elevators, respectively, during 2009 and was also commonly collected in 2010. Other commonly collected insect species included, the red flour beetle, Tribolium castaneum (Herbst); rusty grain beetle, Cryptolestes ferrugineus (Stephens); and sawtoothed grain beetle, Oryzaephilus surinamensis (Linnaeus). Our results showed that failing to clean out boot residual grain and a lack of sanitation of the pit area resulted in high numbers of insects that were transferred through the elevator leg to other locations within a facility. We conclude that the lack of sanitation at elevator and feed mill facilities allowed high numbers of insects to develop.     

Responses of phosphine susceptible and resistant strains of five stored-product insect species to chlorine dioxide

Adults of phosphine susceptible laboratory strains and phosphine resistant field strains of five stored-product insect species were exposed in vials with 0 or 10 g of wheat for different time periods to 0.54 g/m³ (200 ppm) of chlorine dioxide gas. After exposure, adult mortality was determined 5 d later at 28 °C and 65% r.h. The 5-d mortality was 100% in laboratory and field strains of the red flour beetle, Tribolium castaneum (Herbst); sawtoothed grain beetle, Oryzaephilus surinamensis (L.); lesser grain borer, Rhyzopertha dominica (F.); maize weevil, Sitophilus zeamais Motschulsky; and rice weevil, Sitophilus oryzae (L.) that were exposed in vials with 10 g of wheat to chlorine dioxide for 26, 16, 24–34, 18–24, and 15–18 h, respectively. Corresponding exposure durations for these species and strains in vials without wheat were 15, 3, 18–20, 7–15, and 5–7 h, respectively. Dosages of chlorine dioxide producing 99% mortality (LD₉₉) of T. castaneum, O. surinamensis, R. dominica, S. zeamais, and S. oryzae strains in vials with wheat ranged from 14.79−22.57, 8.20–8.41, 15.79–21.60, 10.66–14.53, and 7.67–12.20 g-h/m³, respectively. In vials without wheat, corresponding LD₉₉ values for T. castaneum, R. dominica, and S. zeamais strains were 6.51–8.66, 11.46–23.17, and 5.79–10.26 g-h/m³, respectively. LD₉₉ values for O. surinamensis and S. oryzae could not be computed, because of 100% mortality after a 3–5 h exposure to chlorine dioxide. No adult progeny production of T. castaneum and O. surinamensis was observed after 8 weeks in control and chlorine dioxide-exposed samples. Adult progeny production of Sitophilus spp. was found only in the control samples. The dosage for 99% adult progeny reduction relative to control for R. dominica strains ranged from 10.07 to 18.11 g-h/m³. Chlorine dioxide gas is effective in killing phosphine susceptible and resistant strains of five stored-product insect species and suppressing adult progeny production of three out of the five species.     

Phosphine resistance in stored-product insects collected from various grain storage facilities in Morocco

Despite heavy dependence on phosphine (PH₃) for fumigating stored products, the resistance status of insect pests in Morocco has never undergone a thorough investigation. Some control failures with PH₃ were reported in Morocco, and a previous study showed two field populations of Sitophilus oryzae to be highly resistant to phosphine.We surveyed phosphine resistance in field populations of three major insect pests of stored wheat in Morocco. Around 32% of the samples collected at different storage facilities were found to be infested with one or more species of stored-product beetles. First-generation adult beetles, cultured from the field samples, were subjected to a discriminating dose test for phosphine resistance using an FAO method. The results indicated that, with the exception of one population of S. oryzae, all samples tested contained phosphine-resistant individuals. Treatments at up to 1.8 g m⁻³ of phosphine for 20 h, or at 0.18 g m⁻³ for up to 5 days, indicated that a high degree of resistance was already selected in some of the insect populations.Tests using [³²P]-radiolabelled phosphine showed that the mechanism of resistance in the three insect species tested involved a reduced uptake of the fumigant. The study has highlighted an urgent need for reviewing current fumigation practices in Morocco to ensure effective use of phosphine and avoid further selection of resistance.     

Prevalence,characterization and antibiotic resistance of enterococci from traditional cheeses in Turkey

The aim of the present work is to provide information about Enterococcus strains isolated from traditional Turkish cheese samples in Ankara (Turkey), focusing on their prevalence, phenotypic and genotypic characteristics, and antibiotic resistance. A total of 213 probable enterococcal isolates isolated from 215 samples were identified by phenotypic and genotypic methods. As a result of 16S rDNA sequence analysis, 88 of the 213 enterococci strains were identified as Enterococcus faecium and 125 as Enterococcus faecalis. The E. faecalis strains (58.7%) were identified as the dominant species isolated from cheese samples in Turkey. The 213 Enterococcus strains were tested for susceptibility to 12 different antimicrobial agents. The resistance phenotype were as follow: nalidixic acid (100%), kanamycin (98.6%), rifampicin (78.4%), ampicillin (48.8%), ciprofloxacin (45.5%), erythromycin (18.8%), tetracycline (11.7%), penicillin G (5.6%), chloramphenicol (4.2%), gentamycin (3.8%) and streptomycin (1.4%). None of the strains was resistant to vancomycin. E. faecium strains showed more resistant phenotypes than E. faecalis strains as shown by the antibiotic resistance levels. It was also observed that the resistance of E. faecium and E. faecalis strains against the antibiotics was statistically significant (p ? 0.05). In total, 100% of E. faecium and 88.8% of E. faecalis strains were resistant to multiple drugs.     

A comparison between the impact of sulfuryl fluoride and methyl bromide fumigations on stored-product insect populations in UK flour mills

The impact of sulfuryl fluoride and methyl bromide fumigations upon populations of flour beetles (Tribolium spp.) and of Mediterranean flour moth (Ephestia kuehniella), was evaluated in UK flour mills. Two mills were selected for fumigation with sulfuryl fluoride and two mills for fumigation with methyl bromide. Traps baited with aggregation pheromone lure and an oil-based food attractant were used to monitor populations of rust red flour beetle and confused flour beetle. For the monitoring of Mediterranean flour moth, sticky traps baited with a pheromone lure were used. Traps were placed inside the mill buildings within the areas selected for fumigation. Infestation levels of the insects were monitored for 1–2 weeks prior to fumigation and for a total of 12 weeks post-fumigation. From the calculated percentage reduction in insects trapped per day during the post-fumigation monitoring period it was clear that sulfuryl fluoride fumigations had good efficacy against infestations of Tribolium confusum and E. kuehniella, and compared very favourably with the efficacy of methyl bromide. The absence of T. castaneum in traps pre-fumigation meant that the efficacy of sulfuryl fluoride against this species could not be assessed in this investigation. Indian meal moth, Plodia interpunctella, was also detected in some mills but only post-fumigation.     

A note on the incidence and antibiotic resistance of Staphylococcus aureus isolated from meat and chicken samples

One hundred and fifty samples of raw calf/lamb meat samples (mince and chunks) and chicken parts (giblets, carcass) were analysed for the presence of Staphylococcus aureus. Eighty S. aureus strains were isolated and identified. Resistance of the strains to methicillin and other antibiotics was determined by the Kirby-Bauer disc diffusion test. The overall methicillin resistance rate for S. aureus was 67.5%. Of S. aureus strains, 87.5% were resistant to bacitracin. A high prevalence of penicillin G resistance was detected for S. aureus (53.8%). Few of the strains were resistant to erythromycin (7.5%). All strains were susceptible to vancomycin, sulbactam–ampicillin, ciprofloxacin and cefaperazone–sulbactam. This study confirmed the presence of S. aureus, especially antibiotic-resistant strains, in the foods examined, indicating poor sanitary conditions during processing which may create a health risk for consumers.     

Prevalence and characterization of Enterococcus spp. isolated from Brazilian foods

Enterococci can be used in the food industry as starter or probiotic cultures. However, enterococci are also implicated in severe multi-resistant nosocomial infections. In this study, the prevalence of enterococci in selected Brazilian foodstuffs (raw and pasteurized milk, meat products, cheeses and vegetables) was evaluated. Phenotypic and PCR protocols were used for species identification. Tests for production of gelatinase, haemolysin, bacteriocin and bile salt hydrolysis were done with all enterococci isolates, whereas molecular determination of virulence markers (genes esp, gel, ace, as, efaA, hyl and cylA) and antibiotic resistance was checked only for Enterococcus faecium and Enterococcus faecalis isolates. The antibiotic-resistant isolates were assayed for biofilm formation and adhesion to mammalian cells. From the 120 food samples analyzed, 52.5% were positive for enterococci, meat and cheese being the most contaminated. E. faecium was the predominant species, followed by E. faecalis, E. casseliflavus and Enterococcus gallinarum. Phenotypic tests indicated that 67.7% of isolates hydrolyzed bile salts, 15.2% produced bacteriocin, 12.0% were beta-hemolytic and 18.2% produced gelatinase. Antibiotic resistance (gentamicin, tetracycline and erythromycin) and genes encoding for virulence traits were more frequent in E. faecalis than in E. faecium. Three E. faecium isolates were resistant to vancomycin. Among antibiotic-resistant isolates, 72.4% of E. faecalis were able to form biofilm and 13.8% to adhere to Caco-2 cells. Antibiotic-resistant E. faecalis and E. faecium isolates were grouped by RAPD-PCR and a scattered distribution was noted, indicating that resistance was not related to a particular clone. The spread of virulence/resistance traits in isolates of the two species and different RAPD-types suggest the pathogenic potential of both species. By contrast, the recovery of bacteriocinogenic E. faecium isolates with no virulence traits suggests their potential for biotechnological applications. In conclusion, our results showed that enterococci from Brazilian foods present important dualist aspects for food safety.     

The effects of temperature on flight initiation in a range of moths, beetles and parasitoids associated with stored products

The effects of temperature on flight initiation in a range of stored-product pests and their parasitoids have been studied in laboratory flight chambers. Tests were conducted between 10 and 32.5 °C at intervals of 2.5 °C. The minimum temperatures for flight initiation in the stored-product moths Ephestia kuehniella, E. elutella and Plodia interpunctella were in the range 12.5-15 °C. For the stored-grain beetles, minimum temperatures for flight initiation varied from 17.5 °C for Ahasverus advena and Typhaea stercorea, 20 °C for Rhyzopertha dominica, 25 °C for Tribolium castaneum, to 27.5 °C for Sitophilus oryzae. The minimum temperature for flight initiation in the hymenopteran parasitoids, Anisopteromalus calandrae and Lariophagus distinguendus, was 17.5 °C. Flight is discussed as a factor in sustainable pest management strategies for storage insects; its importance in the spread of infestation and the likely success of physical and biological control methods is highlighted.     

Effect of whole-pea flour and a protein-rich fraction as repellents against stored-product insects

Studies conducted to test the repellency of pea (Pisum sativum L.) products against the red flour beetle, Tribolium castaneum (Herbst), the rice weevil, Sitophilus oryzae (L.) and the lesser grain borer, Rhyzopertha dominica (F.) showed that paddy grains treated with a protein-rich fraction derived from peas var. Bonneville at 1% concentration were repellent to adults. Tribolium castaneum was repelled most quickly followed by S. oryzae and R. dominica.     

Antibiotic resistance genes and identification of staphylococci collected from the production chain of swine meat commodities

Staphylococci harbouring antibiotic resistance (AR) genes may represent a hazard for human health and, as other resistant food-related bacteria, they contribute to the spread of AR. In this study, we isolated resistant staphylococci from an entire swine production chain and investigated the occurrence of 11 genes [aac(6')Ie-aph(2')Ia, blaZ, mecA, vanA, vanB, ermA, ermB, ermC, tet(M), tet(O) and tet(K)] encoding resistance to some antibiotics largely used in clinical practice. The 66 resistant staphylococcal isolates were identified as Staphylococcus epidermidis (27 isolates), Staphylococcus aureus (12), Staphylococcus xylosus (12), Staphylococcus simulans (5), Staphylococcus pasteuri (4), Staphylococcus carnosus (3), Staphylococcus lentus (2) and Staphylococcus sciuri (1). Specific-PCR detection of AR genes showed the prevalence of the tet(K) gene in most of the isolates (89.4%), followed by tet(M) and ermC (about 75%); mecA was detected in more than half of S. aureus and S. epidermidis isolates. The genes vanA and vanB were not retrieved. It was found that a high proportion of coagulase-positive and -negative isolates are multidrug-resistant and some of them carry up to six AR genes. Our findings show that the swine production chain is a source of antibiotic-resistant staphylococci suggesting the importance of resistance surveillance in the food production environment.     

Insecticidal and repellent activity of essential oil from Amomum villosum Lour. and its main compounds against two stored-product insects

The Amomum villosum essential oil was obtained from hydrodistillation and was investigated by GC-MS. The main constituents were bornyl acetate (51.6%), camphor (19.8%), camphene (8.9%) and limonene (6.2%). Insecticidal toxicity of the essential oil was evaluated in this study. It showed that the essential oil possessed contact toxicity against Tribolium castaneum and Lasioderma serricorne (LD₅₀ = 32.4 and 20.4 μg/adult). Three monoterpenoids camphor, camphene and limonene showed strong fumigant toxicity against T. castaneum (LC₅₀ ＜ 2.3, LC₅₀ ＝ 6.2 and 6.2 mg/L air). In addition, repellency of the essential oil was also evaluated. Data showed that the essential oil and all four compounds had repellent activity against T. castaneum and L. serricorne at high concentration (78.63 nL/cm²). But with the decrease of concentration, they showed a different degree of attractant properties.     

Comparison of the desiccation effects of marine and freshwater diatomaceous earths on insects

The desiccating effects of two different types of diatomaceous earths (DEs), of freshwater and marine origin, applied by two different methods, were compared on four species of insects, Cryptolestes ferrugineus, Rhyzopertha dominica, Sitophilus oryzae and Sitophilus granarius. The purpose was to investigate the performance of DEs, including their abrasive and absorptive effects through the accumulation of silica dusts on the insect cuticle, resulting in death of the insects by desiccation. The data obtained showed that similar insects treated with the same type of diatom, but applied either by spraying or dusting, did not have the same initial mortality rate.

After examining and comparing the data obtained, it appeared that weight loss at death of insects subjected to the action of sprayed DEs was slightly less than for insects subjected to dust application, while mortality was slightly more rapid during the observation period. The extra weight of dusted insects was ascribed to greater adherence of dust particles to the treated insects.     

Insecticidal potential of zeolite formulations against three stored-grain insects,particle size effect,adherence to kernels and influence on test weight of grains

Laboratory studies were conducted in order to evaluate the insecticidal potential of three commercially available zeolite formulations against adults of Sitophilus oryzae, Tribolium confusum and Oryzaephilus surinamensis in wheat. For each zeolite formulation, three particle size levels were tested, i.e. 0–50, 0–150 and 0–500 μm. Zeolites were applied at three dose rates, 250, 500 and 1000 ppm, and insect mortality was assessed after 2, 7, 14 and 21 d of exposure. After the final mortality count, dead and alive insects were removed and offspring numbers were determined following an additional period of 65 d. In another series of laboratory bioassays, the effect of zeolite application on the test weight of wheat, maize and barley, as well as the adherence of zeolite particles to wheat, maize, barley and rice kernels was also measured. Oryzaephilus surinamensis was the most susceptible species to zeolite application, regardless of the zeolite formulation, dose and particle size level tested, whereas T. confusum was the most tolerant. No significant differences in efficacy were recorded among the three tested zeolite formulations. At the same time, particle size did not affect zeolite efficacy, at least for the particle size levels tested. All zeolites caused a significant reduction on the test weight of the treated grains. Moreover, zeolite particles showed different adherence among wheat, maize, barley and rice kernels. The results of the present study indicate that zeolites can be used with success as grain protectants, but there is a considerable effect on some physical properties of the grains. This information aims to encourage further evaluation of zeolites as grain protectants.     

Diversity,distribution and antibiotic resistance of Enterococcus spp. recovered from tomatoes,leaves, water and soil on U.S. Mid-Atlantic farms

Antibiotic-resistant enterococci are important opportunistic pathogens and have been recovered from retail tomatoes. However, it is unclear where and how tomatoes are contaminated along the farm-to-fork continuum. Specifically, the degree of pre-harvest contamination with enterococci is unknown. We evaluated the prevalence, diversity and antimicrobial susceptibilities of enterococci collected from tomato farms in the Mid-Atlantic United States. Tomatoes, leaves, groundwater, pond water, irrigation ditch water, and soil were sampled and tested for enterococci using standard methods. Antimicrobial susceptibility testing was performed using the Sensititre microbroth dilution system. Enterococcus faecalis isolates were characterized using amplified fragment length polymorphism to assess dispersal potential. Enterococci (n = 307) occurred in all habitats and colonization of tomatoes was common. Seven species were identified: Enterococcus casseliflavus, E. faecalis, Enterococcus gallinarum, Enterococcus faecium, Enterococcus avis, Enterococcus hirae and Enterococcus raffinosus. E. casseliflavus predominated in soil and on tomatoes and leaves, and E. faecalis predominated in pond water. On plants, distance from the ground influenced presence of enterococci. E. faecalis from samples within a farm were more closely related than those from samples between farms. Resistance to rifampicin, quinupristin/dalfopristin, ciprofloxacin and levofloxacin was prevalent. Consumption of raw tomatoes as a potential exposure risk for antibiotic-resistant Enterococcus spp. deserves further attention.