Phosphine resistance in Tribolium castaneum and Rhyzopertha dominica from stored wheat in Oklahoma

Phosphine gas, or hydrogen phosphide (PH3), is the most common insecticide applied to durable stored products worldwide and is routinely used in the United States for treatment of bulk-stored cereal grains and other durable stored products. Research from the late 1980s revealed low frequencies of resistance to various residual grain protectant insecticides and to phosphine in grain insect species collected in Oklahoma. The present work, which used the same previously established discriminating dose bioassays for phosphine toxicity as in the earlier study, evaluated adults of nine different populations of red flour beetle, Tribolium castaneum (Herbst), and five populations of lesser grain borer, Rhyzopertha dominica (F.) collected from different geographic locations in Oklahoma. One additional population for each species was a laboratory susceptible strain. Discriminating dose assays determined eight out of the nine T. castaneum populations, and all five populations of R. dominica, contained phosphine-resistant individuals, and highest resistance frequencies were 94 and 98%, respectively. Dose-response bioassays and logit analyses determined that LC99 values were approximately 3 ppm for susceptible and 377 ppm for resistant T. castaneum, and approximately 2 ppm for susceptible and 3,430 ppm for resistant R. dominica. The most resistant T. castaneum population was 119-fold more resistant than the susceptible strain and the most resistant R. dominica population was over 1,500-fold more resistant. Results suggest a substantial increase in phosphine resistance in these major stored-wheat pests in the past 21 yr, and these levels of resistance to phosphine approach those reported for other stored-grain pest species in other countries.     

美洲地区主要玉米群体特征及其利用途径分析

美洲地区是全球玉米起源和遗传多样性中心, 优良种质渗入是拓宽我国玉米种质基础的有效途径。通过整理和分析不同学者关于美国、国际玉米小麦改良中心(International Maize and Wheat Improvement Center, CIMMYT)和巴西玉米种质的配合力及杂种优势研究结果, 鉴定出在我国可能有应用潜力的24个优良玉米群体, 包括8个美国群体, 8个CIMMYT群体和8个巴西玉米群体。建议在适应性改良的基础上, 以我国本土化的A群和B群为核心, 将群体BSSS(R)C10、BS10(FR)C14、BS13(S)C9、BSK(HI)C8 Syn 3、BR106、Pop44(C8)、Pop45(C3)与A群种质, 群体BS11(FR)C14、BS16(S)C3 Syn 2、BS29(R)C3、BSCB1(R)C14、BR105、Pop42(C4)与B群种质构建复合群体, 开展改良, 拓宽我国玉米种质类群的遗传基础。     

RESISTANCE TO GRAIN PROTECTANTS AND PHOSPHINE IN COLEOPTEROUS PESTS OF GRAIN STORED ON FARMS IN NEW SOUTH WALES

Tribolium castaneum (Herbst), T. confusum Jacquelin du Val, Sitophilus oryzae (L.), S. granarius (L.), Oryzaephilus surinamensis (L.) and Rhyzopertha dominica (F.) were collected from farms in grain-growing areas in southern and northern N.S.W. and bioassayed for resistance to the grain protectants malathion, fenitrothion, bioresmethrin, carbaryl, pirimiphos-methyl, chlorpyrifos-methyl and the fumigant phosphine.
Results showed that resistance on farms in N.S.W. is similar to resistance reported elsewhere in Australia's grain handling system during the past decade. The only new resistance of practical significance was detected in O. surinamensis where 50% of populations were resistant to fenitrothion (maximum R.F. 68.4x). Fenitrothion resistance was uncommon or not detected in other species. Pirimiphos-methyl resistance was detected in 70% of O. surinamensis populations (maximum RF 44.2x) and in 1 population of T. castaneum (maximum RF 3.1x). Resistance to chlorpyrifos-methyl was detected in 39% of the O. surinamensis populations (maximum RF 8.4x), the only species tested. Low level phosphine resistance was detected in all species (maximum RF 2.2x). Malathion resistance was detected in all species except S. granarius . Resistance to bioresmethrin and carbaryl was not detected in R. dominica , the only species tested.     

Growth and Body Mass Index of Brazilians Ages 9 Through 17 Years

Obesity during adolescence is considered a strong predictor of adult obesity, and obesity and overweight have been increasing among Brazilian adults. To gauge the relative frequency of overweight among adolescents in Brazil, we compared the distributions of body mass index (kg/m²) and stature in national population based samples of the U.S. and Brazil. U.S. adolescents were on average about 10 cm taller than Brazilians, although growth spurts occurred at the same age for both populations. Brazilian adolescents were leaner than their U.S. counterparts. This difference was reduced among girls in the postpubertal period. At age 17 years, U.S. boys were about 10 kg heavier than Brazilian boys, but the difference among girls was only 2 kg. In families above the poverty level in the more developed South region, body mass index distribution for boys was closer to that of the U.S., and older girls tended to have higher body mass index than U.S. girls. Within Brazil, body mass index varied by ethnicity with Mulattos, but not Blacks, of both sexes having lower body mass index than Whites of the same age. Urban adolescents had higher body mass index than those living in rural areas. In general, the patterns seen among Brazilian adults were found among children. Among girls, in particular, overweight has become an identifiable problem during adolescence.     

Population genetics and gene variation in screwworms (Diptera: Calliphoridae) from Brazil

Allozyme variation in New World screwworm,Cochliomyia hominivorax (Coquerel), populations from Brazil was examined. Variability was observed in 8 of 13 enzyme loci and the frequency of the most common allele was <0.95 for seven loci. Observed and expected heterozygosities were 0.159 and 0.165, respectively. Comparisons of the Brazilian populations with previously studied populations from Costa Rica resulted in Nei's genetic distances of between 0.000 and 0.006, with the greatest distance being between populations within Brazil. Comparisons with Mexican populations using only three loci resulted in genetic distances ≤0.031. Goodness-of-fit statistics for Hardy-Weinberg equilibrium and Wright'sF statistics indicated small deviations from expected equilibrium genotype frequencies and low levels of differentiation between populations within Brazil. Differentiation among screwworm populations from Brazil, Costa Rica, and Mexico was minimal.     

Use of a web-based model for aeration management in stored rough rice

A web-based model was used to simulate the impact of aeration on population growth of the lesser grain borer, Rhyzopertha dominica (F.), and the rice weevil, Sitophilus oryzae (L.), in stored rough rice, Oryza sativa L., at Beaumont, TX. Simulations were run for each of 10 yr with 1 August as the start date; 31 December as the end date; beginning populations of 2.5 adults per metric ton (1000 kg); starting grain temperatures of 29.4, 32.2, and 35.0 degrees C; and aeration airflow rates of 0.27, 0.79, and 1.40 m3/min/metric ton of rice. In the absence of aeration, populations of both species increased exponentially, with maximum production of R. dominica and S. oryzae at starting grain temperatures of 35.0 and 32.2 degrees C, respectively. Final predicted populations of R. dominica on 31 December from grain starting temperatures of 29.4, 32.2, and 35.0 degrees C were 5465, 6848, and 11,855 per ton, respectively; final predicted populations of S. oryzae were 13,288, 21,252 and 4355, respectively. Aeration led to a reduction in grain temperature and a decrease in pest populations, regardless of starting grain temperature or aeration airflow rates. Predicted populations of R. dominica on 31 December ranged from 12 to 63 adults per ton at all grain starting temperatures and airflow rates; populations of S. oryzae on 31 December ranged from 108 to 193 adults per ton at all grain starting temperatures and airflow rates. The predicted population levels in aerated rice represented at least a 98% reduction compared with unaerated rice. Results show the utility of the web-based model and how the various model inputs can help define broader patterns of insect control in rice stored in the south central United States.     

Genetic variation and structure in native populations of the fire ant Solenopsis invicta: evolutionary and demographic implications

We studied population genetic variation and structure in the fire ant Solenopsis invicta using nuclear genotypic and mitochondrial DNA (mtDNA) sequence data obtained from samples collected throughout its native range. Geographic populations are strongly differentiated at both genomes, with such structure more pronounced in Brazil than in Argentina. Higher-level regional structure is evident from the occurrence of isolation-by-distance patterns among populations, the recognition of clusters of genetically similar, geographically adjacent populations by ordination analysis, and the detection of an mtDNA discontinuity between Argentina and Brazil coinciding with a previously identified landform of biogeographical relevance. Multiple lines of evidence from both genomes suggest that the ancestors of the ants we studied resembled extant northern Argentine S. invicta , and that existing Brazilian populations were established more recently by serial long-distance colonizations and/or range expansions. The most compelling evidence for this is the corresponding increase in F _K (a measure of divergence from a hypothetical ancestor) and decrease in genetic diversity with distance from the Corrientes population in northern Argentina. Relatively deep sequence divergence among several mtDNA clades, coupled with geographical partitioning of many of them, suggests prolonged occupation of South America by S. invicta in more-or-less isolated regional populations. Such populations appear, in some cases, to have come into secondary contact without regaining the capacity to freely interbreed. We conclude that nominal S. invicta in its native range comprises multiple entities that are sufficiently genetically isolated and diverged to have embarked on independent evolutionary paths.  © 2007 The Linnean Society of London, Biological Journal of the Linnean Society , 2007, 92 , 541–560.     

Microsatellite Markers Reveal Genetic Variation within Sclerotinia sclerotiorum Populations in Irrigated Dry Bean Crops in Brazil

Microsatellites are powerful markers to infer population genetic parameters. We used 10 microsatellite loci to characterize the genetic diversity and structure of 79 samples of Sclerotinia sclerotiorum isolated from four Brazilian dry bean populations and observed that eight of them were polymorphic within populations. We identified 102 different haplotypes ranging from 6 to 18 per locus. Analyses based on genetic diversity and fixation indices indicated variability among and within populations of 28.79% (F_ST = 28793) and 71.21%, respectively. To examine genetic relatedness among S. sclerotiorum isolates, we used internal spacer (ITS1‐5.8S‐ITS2) restriction fragment length polymorphism (PCR‐RFLP) and sequencing analysis. PCR‐RFLP analysis of these regions failed to show any genetic differences among isolates. However, we detected variability within the sequence, which does not support the hypothesis of clonal populations within each population. High variability within and among populations may indicate the introduction of new genotypes in the areas analysed, in addition to the occurrence of clonal and sexual reproduction in the populations of S. sclerotiorum in the Brazilian Cerrado.     

The rph2 gene is responsible for high level resistance to phosphine in independent field strains of Rhyzopertha dominica

The lesser grain borer Rhyzopertha dominica (F.) is one of the most destructive insect pests of stored grain. This pest has been controlled successfully by fumigation with phosphine for the last several decades, though strong resistance to phosphine in many countries has raised concern about the long term usefulness of this control method. Previous genetic analysis of strongly resistant (SR) R. dominica from three widely geographically dispersed regions of Australia, Queensland (SR(QLD)), New South Wales (SR(NSW)) and South Australia (SR(SA)), revealed a resistance allele in the rph1 gene in all three strains. The present study confirms that the rph1 gene contributes to resistance in a fourth strongly resistant strain, SR2(QLD), also from Queensland. The previously described rph2 gene, which interacts synergistically with rph1 gene, confers strong resistance on SR(QLD) and SR(NSW). We now provide strong circumstantial evidence that weak alleles of rph2, together with rph1, contribute to the strong resistance phenotypes of SR(SA) and SR2(QLD). To test the notion that rph1 and rph2 are solely responsible for the strong resistance phenotype of all resistant R. dominica, we created a strain derived by hybridising the four strongly resistant lines. Following repeated selection for survival at extreme rates of phosphine exposure, we found only slightly enhanced resistance. This suggests that a single sequence of genetic changes was responsible for the development of resistance in these insects.     

Genetic diversity and assessment of 23 microsatellite markers for parentage testing of Santa Inês hair sheep in Brazil

Santa Inês is the most common hair sheep breed in Brazil and probably has the highest genetic diversity among sheep breeds in this country. Successful breeding programs for Brazilian sheep breeds are not common for various reasons, including a lack of control of parentage in the flocks. We developed an allele frequency database for 23 STR loci for the Santa Inês breed based on 285 animals sampled from five populations distributed across the central-western and north-eastern regions of Brazil. The marker set included seven microsatellites used in the 2011 International Society for Animal Genetics sheep genotyping comparison tests and all eight microsatellites currently approved by the Brazilian Agricultural Ministry laboratory accreditation guidelines for sheep identification. The microsatellites had an average of 10 alleles and a mean expected heterozygosity of 0.745. Combined paternity exclusion probabilities when no parent or one parent was known were >99.99%. A small proportion (5.8%) of the existing genetic variation was found to be among the Santa Inês populations, possibly derived from genetic drift and selection. We found that the marker panel proposed by the Agricultural Ministry, although generally useful, should be enhanced by including more markers for improved exclusionary power in parentage testing. This database provides a useful tool for parentage testing of this major Brazilian breed, contributing to improved management and breeding of existing herds.     

Frequency and origins of hemoglobin S mutation in African-derived Brazilian populations

Africans arrived in Brazil as slaves in great numbers, mainly after 1550. Before the abolition of slavery in Brazil in 1888, many communities, called quilombos, were formed by runaway or abandoned African slaves. These communities are presently referred to as remnants of quilombos, and many are still partially genetically isolated. These remnants can be regarded as relicts of the original African genetic contribution to the Brazilian population. In this study we assessed frequencies and probable geographic origins of hemoglobin S (HBB*S) mutations in remnants of quilombo populations in the Ribeira River valley, S?o Paulo, Brazil, to reconstruct the history of African-derived populations in the region. We screened for HBB*S mutations in 11 quilombo populations (1,058 samples) and found HBB*S carrier frequencies that ranged from 0% to 14%. We analyzed beta-globin gene cluster haplotypes linked to the HBB*S mutation in 86 chromosomes and found the four known African haplotypes: 70 (81.4%) Bantu (Central Africa Republic), 7 (8.1%) Benin, 7 (8.1%) Senegal, and 2 (2.3%) Cameroon haplotypes. One sickle cell homozygote was Bantu/Bantu and two homozygotes had Bantu/Benin combinations. The high frequency of the sickle cell trait and the diversity of HBB*S linked haplotypes indicate that Brazilian remnants of quilombos are interesting repositories of genetic diversity present in the ancestral African populations.     

Genetic structure in Brazilian breeding colonies of the Roseate Spoonbill (Platalea ajaja, Aves: Threskiornithidae)

Roseate Spoonbills (Platalea ajaja, Linnaeus) are wading birds present in two of the most important Brazilian wetlands: the Pantanal wetlands and Rio Grande do Sul marshes. Natural populations of these species have not been previously studied with variable nuclear molecular markers. In order to support decision making regarding the management and conservation of these populations, we estimated and characterized the distribution of genetic variability among five Brazilian breeding colonies. The average observed heterozygosity in Brazilian Roseate Spoonbill populations (Ho = 0.575) did not differ significantly from the value determined in a U.S. wild-caught sample of 15 individuals, using data generated by the same set of microsatellite loci. Considering that the U.S. population underwent a recent reduction in size, we discuss this result supposing that the U.S. population was not genetically affected or that both populations had suffered a bottleneck. Global F(ST) indicated the lack of genetic differentiation among colonies, indicating the occurrence of past and/or present gene flow among them. Analysis of molecular variance revealed that most of the genetic variation is distributed within the colonies. Results are explained by a recent origin of colonies or by high levels of gene flow. Management decisions should take into consideration the fact that, even in the presence of high genetic exchange, ecological adaptations to different environments are important for species survival.     

Is there genetic structure in populations of Helicoverpa armigera from Australia?

Recent studies suggest that populations of the pest moth Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) may be genetically differentiated over short distances and time periods within Queensland, Australia. To test for genetic structure in another region of Australia, we characterized population differentiation in Victorian samples of H. armigera using eight microsatellite loci. We found no evidence of genetic structure among samples from different locations or among samples collected at different times. Moreover, Victorian samples were not differentiated from other samples of H. armigera from Queensland and New Zealand. All samples showed substantial deviations from Hardy–Weinberg equilibrium, suggesting a high frequency of null alleles typically found in microsatellites of Lepidoptera. These results indicate that populations of H. armigera are not strongly structured among regions in south‐eastern Australia.     

Molecular dissimilarities of <Emphasis Type="Italic">Rhipicephalus sanguineus</Emphasis> (Acari: Ixodidae) in Brazil and its relation with samples throughout the world: is there a geographical pattern?

In this study the genetic variability of Rhipicephalus sanguineus within Brazil and its relation with ticks of the same group from different continents was evaluated. Mitochondrial 12S and 16S rDNA fragments of R. sanguineus from seven Brazilian States were sequenced and compared to GenBank sequences of R. sanguineus and R. turanicus ticks from Africa, Asia, Europe, South America and USA. Results indicate a relatively high intra-specific variability between Brazilian samples but also a global latitude linked distribution pattern of at least two major R. sanguineus groups; one group distributed from latitude 25°N to 22°S including R. sanguineus from Brazil, Taiwan and Thailand and R. turanicus from Zambia and Zimbabwe, and the other group found closer to the poles, roughly above 29°N and below 30°S with ticks from Argentina, Uruguay, France, Oklahoma (USA), Israel and Egypt.     

Genetic structure and molecular variability of Spodoptera frugiperda (Lepidoptera: Noctuidae) collected in maize and cotton fields in Brazil

The purpose of this research was to evaluate the genetic similarity and structure of the fall armyworm, Spodoptera frugiperda (J.E. Smith), populations associated with maize and cotton crops in Brazil using amplified fragment length polymorphisms. Mean genetic similarity among populations was 0.45. The unweighted pair group method with arithmetic mean analysis dendrograms did not separate populations of S. frugiperda into clusters related to the host plant in which the insects were collected. No genetic variation was observed among maize and cotton populations of S. frugiperda, suggesting that the same populations are injuring both crops in Brazil. This research validates the need for stewardship of crop-protection methods for managing S. frugiperda to reduce the incidence of pesticide resistance, due to the spatial and temporal overlapping of maize and cotton crops in some regions in Brazil.     

Population genetic structure of mahogany (Swietenia macrophylla King, Meliaceae) across the Brazilian Amazon, based on variation at microsatellite loci: implications for conservation

Mahogany (Swietenia macrophylla, Meliaceae) is the most valuable and intensively exploited Neotropical tree. No information is available regarding the genetic structure of mahogany in South America, yet the region harbours most of the unlogged populations of this prized hardwood. Here we report on the genetic diversity within and the differentiation among seven natural populations separated by up to 2100 km along the southern arc of the Brazilian Amazon basin. We analysed the variation at eight microsatellite loci for 194 adult individuals. All loci were highly variable, with the number of alleles per locus ranging from 13 to 27 (mean = 18.4). High levels of genetic diversity were found for all populations at the eight loci (mean HE = 0.781, range 0.754-0.812). We found moderate but statistically significant genetic differentiation among populations considering both estimators of FST and RST, theta = 0.097 and rho = 0.147, respectively. Estimates of theta and rho were significantly greater than zero for all pairwise population comparisons. Pairwise rho-values were positively and significantly correlated with geographical distance under the isolation-by-distance model. Furthermore, four of the populations exhibited a significant inbreeding coefficient. The finding of local differentiation among Amazonian mahogany populations underscores the need for in situ conservation of multiple populations of S. macrophylla across its distribution in the Brazilian Amazon. In addition, the occurrence of microgeographical genetic differentiation at a local scale indicates the importance of maintaining populations in their diverse habitats, especially in areas with mosaics of topography and soil.     

Immediate and delayed mortality of Rhyzopertha dominica (Coleoptera: Bostrichidae) and Sitophilus oryzae (Coleoptera: Curculionidae) adults exposed to spinosad-treated commodities

A series of tests was conducted to characterize differences in the mortality of the lesser grain borer, Rhyzopertha dominica (F.) (Coleoptera: Bostrichidae), and rice weevil, Sitophilus oryzae (L.) (Coleoptera: Curculionidae), exposed to three commodities treated with a liquid and dry spinosad formulation. In laboratory bioassays, adults of the two insect species were exposed to untreated wheat, Triticum aestivum L., corn, Zea mays L., and sorghum, Sorghum bicolor (L.) Moench., and to commodities treated with 1 mg (AI)/kg of liquid and dry spinosad formulations. Mortality was assessed from independent samples examined at specific time intervals to determine immediate mortality and after 24 h of recovery on untreated grain at 28 degrees C and 65% RH to determine delayed mortality. Comparison of the time required for 50% (LT50) and 95% (LT95) mortality indicated that R. dominica adults were consistently and significantly more susceptible (died quickly) than S. oryzae adults when exposed to spinosad-treated commodities. In general, the toxicity of liquid and dry spinosad formulations was similar against R. dominica or S. oryzae. The toxicity of spinosad to each species varied slightly among the three commodities, and there were no consistent trends to suggest that spinosad was more effective on one commodity versus another. LT50 values based on immediate mortality for R. dominica on all commodities ranged from 0.45 to 0.74 d; corresponding values based on delayed mortality ranged from 0.04 to 0.23 d, suggesting delayed toxic action of spinosad in R. dominica. LT50 values based on immediate and delayed mortality for S. oryzae on all three commodities treated with the two spinosad formulations were essentially similar and ranged from 2.75 to 4.56 d. LT95 values for R. dominica based on immediate mortality on spinosad-treated commodities ranged from 1.75 to 3.36 d, and those based on delayed mortality ranged from 0.49 to 1.88 d. There were no significant differences in LT95 values based on immediate and delayed mortality for S. oryzae on spinosad-treated commodities, and the LT95 values ranged from 7.62 to 18.87 d. The toxicity of spinosad was enhanced during a 24-h holding period after removal from spinosad-treated commodities only against R. dominica adults, and possible reasons for increased postexposure mortality of R. dominica adults after brief exposures to spinosad warrant further study.     

Differences in population structure of the anther smut fungus Microbotryum violaceum on two closely related host species, Silene latifolia and S. dioica

We investigated the genetic population structure of the sexually transmitted plant pathogen, the fungus Microbotryum violaceum, on the two closely related host species Silene latifolia and S. dioica using microsatellite markers. We found strong deviations from Hardy-Weinberg expectations, with significant heterozygote deficiency in almost all populations. Fungal strains from the two host species were differentiated, and these host races differed in amount of variation within populations and differentiation among populations. Anther smut from S. latifolia harboured significantly less microsatellite diversity and were more genetically differentiated from each other than those from S. dioica. Small effective population sizes, rapid population turnover, and less gene flow among populations could lead to this higher population differentiation and lower within population genetic diversity for anther smut populations on S. latifolia than on S. dioica. These results are in concordance with host ecology because S. latifolia grows in more disturbed habitats than S. dioica and may provide a shorter-lived host environment.     

Analysis of Amblyomma sculptum haplotypes in an area endemic for Brazilian spotted fever

Amblyomma sculptum (Ixodida: Ixodidae) Berlese, 1888, a member of the Amblyomma cajennense complex, is the major vector of Brazilian spotted fever (BSF) in southeastern Brazil. In this study, the genetic diversity of A. sculptum populations in the state of Rio de Janeiro (RJ), Brazil, was investigated because genetic variability in tick populations may be related to vector competence. Samples of A. sculptum from 19 municipalities in 7 regions of RJ were subjected to DNA extraction, amplification and sequencing of D‐loop, cytochrome oxidase II and 12S rDNA mitochondrial genes. These sequences were used to map the genetic diversity of this tick. Amblyomma sculptum populations are genetically diverse in RJ, especially in the South Centre and Highland regions. Few unique haplotypes were observed in all populations, and the majority of genetic variation found was among ticks within each population. Phylogenetic reconstruction reinforced the assumption that all the haplotypes identified in RJ belong to A. sculptum. However, some RJ haplotypes are closer to A. sculptum from Argentina than to A. sculptum from elsewhere in Brazil. In RJ, A. sculptum has high genetic diversity, although little genetic differentiation. Observations also indicated a high level of gene flow among the studied populations and no evidence of population structure according to region in RJ.     

Detection of stored-grain insect infestation in wheat transported in railroad hopper-cars

Levels of insect infestation, insect spatial distribution, and the relationship between the number of insect-damaged kernels (IDK) and the number of insects present in grain samples in three-hopper railcars transporting wheat from country elevators to a mill were studied. Six of eight sampled railcars were infested with more than two species of insects. The most abundant species collected were the lesser grain borer, Rhyzopertha dominica (F.), and rusty grain beetle, Cryptolestes ferrugineus (Stephens), with the larval stage of the two species being the most prevalent (>90%). The spatial distributions of these two species within the grain mass were typically clumped in railcar compartments containing >0.4 insect/2.75-kg sample of wheat, and these foci of high-infestation levels varied in compartments within the railcars and among the sampled railcars. There were no significant correlations between IDK and insect density for any of the different stage-specific insect populations that were collected in the grain samples. Mean numbers of immatures and IDK differed among railcars and compartments within railcars, but not among grain depths. Number of insects in the first discharge sample was not correlated with mean numbers of insects in the entire compartment. This indicates that each compartment of a railcar should be sampled to determine level of insect infestation but that sampling at different depths within a compartment is less important.