African Citrus Greening Disease in East Africa: Incidence, Severity, and Distribution Patterns.

African citrus greening disease (ACGD) is considered as one of the major diseases of citrus threatening citrus production in East Africa. Our study aimed for the first time to assess the incidence, severity, and distribution patterns of ACGD in Kenya and Tanzania. In total, 105 citrus orchards were assessed in 13 regions representing low, mid, and high altitude areas. In each backyard and orchard, trees were randomly selected and rated for visual ACGD symptoms; then leaves and insect samples collected for analysis of 'Candidatus Liberibacter africanus' (CLaf), the presumptive causal agent of ACGD. Endpoint PCR, sequencing, and molecular phylogenetic tools were employed to confirm the identity of potential circulating pathogens. Incidence and severity of ACGD varied significantly among the different regions. Both Trioza erytreae (Del Guerico) (Hemiptera: Triozidae) and the invasive Asian citrus psyllid vector Diaphorina citri (Kuwayama) (Hemiptera: Liviidae) were found to co-occur in upper and lower midland regions. Molecular characterization identified 'Candidatus Liberibacter africanus spp. Clausenae' (CLafCl) as the main causal agent of ACGD in most of the citrus plants and insect samples. No instances of Candidatus Liberibacter asiaticus infection were found. These findings provide valuable insights into understanding and management of ACGD by employing stringent and early disease detection tools to curb the spread of the disease.

DNA Barcode Reference Library for the African Citrus Triozid, Trioza erytreae (Hemiptera: Triozidae): Vector of African Citrus Greening.

Citrus (Citrus spp.) production continues to decline in East Africa, particularly in Kenya and Tanzania, the two major producers in the region. This decline is attributed to pests and diseases including infestation by the African citrus triozid, Trioza erytreae (Del Guercio) (Hemiptera: Triozidae). Besides direct feeding damage by adults and immature stages, T. erytreae is the main vector of 'Candidatus Liberibacter africanus', the causative agent of Greening disease in Africa, closely related to Huanglongbing. This study aimed to generate a novel barcode reference library for T. erytreae in order to use DNA barcoding as a rapid tool for accurate identification of the pest to aid phytosanitary measures. Triozid samples were collected from citrus orchards in Kenya, Tanzania, and South Africa and from alternative host plants. Sequences generated from populations in the study showed very low variability within acceptable ranges of species. All samples analyzed were linked to T. erytreae of GenBank accession number KU517195. Phylogeny of samples in this study and other Trioza reference species was inferred using the Maximum Likelihood method. The phylogenetic tree was paraphyletic with two distinct branches. The first branch had two clusters: 1) cluster of all populations analyzed with GenBank accession of T. erytreae and 2) cluster of all the other GenBank accession of Trioza species analyzed except T. incrustata Percy, 2016 (KT588307.1), T. eugeniae Froggatt (KY294637.1), and T. grallata Percy, 2016 (KT588308.1) that occupied the second branch as outgroups forming sister clade relationships. These results were further substantiated with genetic distance values and principal component analyses.

A Mixed Infection of Lettuce chlorosis virus, Papaya ringspot virus, and Tomato yellow leaf curl virus-IL Detected in a Texas Papaya Orchard Affected by a Virus-Like Disease Outbreak.

Severe virus-like symptoms consisting of mosaic, distortion, yellowing, and brittleness were observed on papaya plants in a 20-ha orchard in South Texas during the 2014-15 growing season. Incidence of symptomatic plants increased from ∼40 to 100% within 6 months of the outbreak; the most severely affected plants were stunted, and fruit yield and quality were reduced compared with asymptomatic plants. The orchard papaya plant virome was explored using the Illumina NextSeq 500 platform and results were validated by Sanger DNA sequencing of complete viral genomes obtained by PCR amplification. The combined results revealed the presence of Papaya ringspot virus (PRSV; Potyvirus), Lettuce chlorosis virus (LCV; Crinivirus), and Tomato yellow leaf curl virus-IL (TYLCV-IL; Begomovirus). The RT-PCR analyses of leaves from 51 randomly sampled papaya plants indicated the presence of PRSV, LCV, and TYLCV-IL in 100, 39.2, and 15.7% of the samples, respectively. Plants infected with PRSV, in combination with LCV and/or TYLCV-IL, exhibited more severe symptoms compared with plants infected with PRSV alone. Furthermore, successful whitefly-mediated transmission of TYLCV-IL and LCV was accomplished by exposing virus-free papaya seedlings to viruliferous Bemisia tabaci (Genn.) under greenhouse conditions. The results of this study document a new host record for LCV and the first successful whitefly-mediated transmission of TYLCV-IL and LCV to papaya. As a perennial crop, infected papaya serving as an over-seasoning reservoir for TYLCV-IL and LCV, presents a new challenge to viral disease management in papaya orchards.

Efficacy and uptake of soil-applied imidacloprid in the control of Asian citrus psyllid and a citrus leafminer, two foliar-feeding citrus pests.

The systemic neonicotinoid insecticide imidacloprid, Admire Pro, was applied to 3- and 4-yr-old nonbearing 'Rio Red' grapefruit, Citrus x paradisi Macfad., trees in 2006 and 2007, respectively, to determine its effects in the control of two major citrus pests, the Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Psyllidae), and a citrus leafminer Phyllocnistis citrella Stainton (Lepidoptera: Gracillariidae). Young flush shoots were randomly collected weekly for 13 and 11 wk in 2006 and 2007, respectively, to determine the infestation levels and densities of immature stages of both Asian citrus psyllid and P. citrella. Additional flush shoot samples were collected in 2007 and titers of imidacloprid in leaf tissue were determined using an enzyme-linked immunosorbent assay. Soil application of imidacloprid significantly reduced the infestation levels and densities of both pests on flush shoots, starting from the second week post application. The effects of the neonicotinoid insecticide were similar in both years. Analysis of imidacloprid concentration in leaf tissue showed a gradual increase during the first 3 wk, and titers remained well above 200 ppb for 11 wk postapplication. Significant positive correlations were obtained between imidacloprid titers in leaf tissue and the percentage of control levels achieved for both pests. A high level of suppression of both P. citrella and Asian citrus psyllid populations on citrus trees was associated with imidacloprid titer in leaf tissue >200 ppb, which was reached 2 wk after soil treatment. Although soil application of imidacloprid did not provide rapid knockdown of Asian citrus psyllid and P. citrella populations, it resulted in chronic residues in leaf tissue and long-term suppression of both pests.

Responses of the Asian citrus psyllid to volatiles emitted by the flushing shoots of its rutaceous host plants.

Diaphorina citri Kuwayama (Hemiptera: Psyllidae) carries Candidatus liberibacter spp., the putative causal agents of Huanglongbing. D. citri reproduces and develops only on the flushing shoots of its rutaceous host plants. Here we examined whether D. citri is attracted to host plant odors and a mixture of synthetic terpenes. Tests conducted in a vertically oriented Y-tube olfactometer showed that both males and females preferentially entered the Y-tube arm containing the odor from the young shoots of Murraya paniculata (L.) Jack and Citrus limon L. Burm. f. cultivar Eureka. Only males exhibited a preference for the odor of C. sinensis L., whereas the odor of C. x paradisi MacFadyen cultivar Rio Red was not attractive to both sexes. The volatiles emitted by young shoots of grapefruit cultivar Rio Red, Meyer lemon (Citrus x limon L. Burm.f.), and M. paniculata were analyzed by gas chromatograph-mass spectrometry. The samples were comprised of monoterpenes, monoterpene esters, and sesquiterpenes. The number of compounds present varied from 2 to 17, whereas the total amount of sample collected over 6 h ranged from 5.6 to 119.8 ng. The quantitatively dominant constituents were (E)-beta-ocimene, linalool, linalyl acetate, and beta-caryophyllene. The attractiveness of a mixture of synthetic terpenes, modeled on the volatiles collected from M. paniculata, was evaluated in screened cages in a no-choice test. At three observation intervals, significantly more individuals were trapped on white targets scented with the mixture than on unscented targets. These results indicate the feasibility of developing D. citri attractants patterned on actual host plant volatiles.

Effects of photoperiod on boll weevil (Coleoptera: Curculionidae) development, survival, and reproduction.

Effects of photoperiod on development, survival, feeding, and oviposition of boll weevils, Anthonomus grandis grandis Boheman, were assessed under five different photophases (24, 14, 12, 10, and 0 h) at a constant 27 degrees C temperature and 65% RH in the laboratory. Analyses of our results detected positive relationships between photoperiod and puncturing (mean numbers of oviposition and feeding punctures per day), and oviposition (oviposition punctures/oviposition+feeding punctures) activities, and the proportion of squares attacked by boll weevil females. When boll weevil females developed in light:darkness cycles, they produced a significantly higher percentage of eggs developing to adulthood than those developed in 24-h light or dark conditions. In long photoperiod (24:0 and 14:10 h), the number of female progeny was significantly higher and their development time was significantly shorter than those developed in short photoperiod (0:24 and 10:14 h). Lifetime oviposition was significantly highest at 12- and 14-h photophase, lowest at 0- and 10-h photophase, and intermediate at 24 h of light. Life table calculations indicated that boll weevil populations developed in a photoperiod of 14:10 and 12:12 (L:D) h will increase an average of two-fold each generation (Ro) compared with boll weevils developed in 24:0- and 10:14-h photoperiods and 15-fold compared with those at 0:24 h. Knowledge of the photoperiod-dependent population growth potential is critical for understanding population dynamics to better develop sampling protocols and timing insecticide applications.

Reproductive potential of overwintering, F1, and F2 female boll weevils (Coleoptera: Curculionidae) in the Lower Rio Grande Valley of Texas.

The feeding and oviposition activity of overwintering boll weevils, Anthonomus grandis grandis (Boheman), and seasonal fluctuations in development, survival, and reproduction of progeny of overwintering and first- and second-generation boll weevil females were determined in the laboratory at 27 degrees C, 65% RH, and a photoperiod of 12:12 (L:D) h. During the cotton-free period in the Lower Rio Grande Valley, female boll weevils without access to cotton resorb their unlaid eggs and enter reproductive diapause. However, when they were provided daily with greenhouse-grown cotton squares, commencement of oviposition began after 7, 15, or 20 d, depending on when they were captured. Females captured later in the winter fed longer before laying eggs than those captured in the early fall, suggesting that it may take females longer to terminate diapause the longer they have been dormant. The rate of feeding by females was significantly less during the winter months, and this may have affected the rate of diet-mediated termination of dormancy. Females of the first and second generations after the overwintering generation produced a significantly higher percentage of progeny surviving to adulthood and a higher proportion of these progeny were females. Offspring development time from overwintering female parents was significantly longer than that from first and second generations under the same laboratory conditions. The total number of lifetime eggs produced by females of the second generation during the cotton-growing season were approximately 9.9-fold higher than for overwintering females and 1.5-fold higher than for first-generation females. Life table calculations indicated that the population of second-generation boll weevils increased an average of 1.5-fold higher each generation than for females of the first generation and 22.6-fold higher than for overwintering females. Our data showed variation in boll weevil survival, development, and reproductive potential among the overwintering and first- and second-generation females, suggesting inherent seasonal fluctuations in these parameters.

Olfactory and visual stimuli affecting host plant detection in Homalodisca coagulata (Hemiptera: Cicadellidae).

The relative effects of visual and olfactory stimuli on host plant detection in immature and adult Homalodisca coagulata Say (Homoptera: Cicadellidae) were studied using a novel olfactometer and factorial experimental designs. Colored, gray, and white cards were used as visual targets. Each card was attached to a glass thistle tube from which host-plant odor (from Vigna unguiculata L.) or blank, humidified air was dispensed. Visual + odor stimuli combinations were presented in no-choice tests. Nymphs were released onto a perch stick downwind from the target. Nymph response to color + odor treatments was measured by the duration of orientation behavior, residence time on the perch, and percentage of individuals that jumped to the target. The assay was modified so that adults crawled from the perch onto the target. Adult response was measured by the duration of individual behaviors (e.g., foraging) and by their position and residence time on the target. Both main effects and interactive effects of the stimuli were observed. Nymphs showed a decrease in orientation and residence times in the colored target + host odor treatments and increased jumping response in the gray + host odor treatment. When adults were exposed to host odor, the duration of foraging behavior increased, whereas crawling and phototactic behaviors decreased. Although nymphs and adults responded to visual stimuli + blank air treatments, host odor enhanced their responses. The primary effect of host odor on host detection behavior may be to enhance H. coagulata responsiveness to visual cues.

Size-dependent feeding and reproduction by boll weevil (Coleoptera: Curculionidae).

The considerable variation in adult size of the boll weevil, Anthonomus grandis grandis Boheman, has been well documented, but the influences of adult size on reproductive rate are not known. We examined the relationship between the size of boll weevils and their feeding and oviposition. Weevils weighed to the nearest milligram were grouped into five categories based on pupal weight: < or =5, 6-10, 11-15, 16-20, and >20 mg. Numbers of lifetime punctures produced in flower buds (squares) of cotton, Gossypium hirsutum L., by both sexes of adults tended to increase with pupal weight. Boll weevil females with pupal weights >10 mg produced progeny with significantly higher survival to adulthood and also produced a higher percentage of female progeny than those with pupal weights < or =10 mg. The population growth indices for females having pupal weights >10 mg averaged 1.8-fold higher than those of females weighing < or =10 mg. Survivorship of adults of both sexes also tended to increase with pupal weight. The percentage of females laying eggs on any given day averaged 2.1 times higher when their pupal weights were >10 mg than when their pupal weights were < or =10 mg. Although small size negatively affected female reproductive potential, even extremely small females produced some viable offspring. However, the penalties of small adult size, in terms of longevity and reproductive potential, suggest that cultural practices that result in the production of small adults may be used to impact weevil populations.

Leptodictya tabida (Hemiptera: Tingidae): a potential threat to sugarcane production in Lower Rio Grande Valley of Texas.

Areawide surveys and replicated cultivar trials were conducted in 2001 and 2002 in sugarcane (Saccharum spp.) fields in the Lower Rio Grande Valley of Texas to assess the distribution and incidence of the sugarcane tingid Leptodictya tabida (Herrich-Schaeffer). L. tabida was found in all fields surveyed during both years, infesting 60 and 68% of the plants, respectively. The average percentage of leaves infested was 11% in 2001 and 15% in 2002. In 2001, 'CP70-1133' was the most infested, 'CP72-1210' was the least infested, and intermediate infestation levels were evident in 'CP70-321' and 'TCP87-3388'. In 2002, however, TCP87-3388 and CP70-321 were more heavily infested, and CP71-1240 and CP71-1405 were the least infested. Mean densities of L. tabida recovered per plant varied between 1.2 bugs on CP72-1210 and 5.1 on CP70-1133 in 2001, and in 2002, from zero bugs on CP71-1240 and CP71-1405 to 5.3 on CP72-1210. In the cultivar trials, cultivar differences also were evident in both plant and leaf infestation levels, and the proportion of immatures to total L. tabida populations; 'HoCP91-555' had the lowest L. tabida infestations and 'NCo-310' had the greatest levels in both years. Although >5000 L. tabida from the field were collected and kept in the laboratory, no parasitoids were found. The distribution of the infestations during the surveys and in the field trial evaluations suggested that L. tabida populations have been spreading in sugarcane across the Lower Rio Grande Valley. Potential varietal resistance mechanisms are discussed.

Systemic effects of neem on western flower thrips, Frankliniella occidentalis (Thysanoptera: Thripidae).

The systemic effects of neem on the western flower thrips, Frankliniella occidentalis (Pergande), were investigated in laboratory trials using green bean, Phaseolus vulgaris L., in arena and microcosm experiments. In arena experiments, systemic effects of neem against western flower thrips larvae on primary bean leaves were observed with maximum corrected mortality of 50.6%. In microcosm experiments using bean seedlings, higher efficacy in the control of western flower thrips were observed with soil applications of neem on a substrate mixture (i.e., Fruhstorfer Erde, Type P, and sand) in a 1:1 ratio (93% corrected mortality) compared with application on the commercial substrate only (76% corrected mortality). However, longer persistence of neem was observed with soil application on the commercial substrate, which showed effects against thrips for up to 6 d after application. In addition to systemic effects observed on all foliage-feeding stages of western flower thrips, mortality on contact and repellent effects were observed on soil-inhabiting stages after soil applications of neem. Finally, bean seedlings grown from seeds pregerminated for 3 d in neem emulsion were also toxic to western flower thrips.

Effects of snowdrop lectin on Mexican rice borer (Lepidoptera: Pyralidae) life history parameters.

The effects of the snowdrop lectin, Galanthus nivalis agglutinin (GNA), delivered through an artificial diet, on growth, development, and life history parameters of the Mexican rice borer, Eoreuma loftini (Dyar), were evaluated in the laboratory. Incorporation of GNA at three treatment levels, 0.5, 1.0, and 2.0% of total dietary protein, in the larval diet significantly decreased larval survivorship and percentage of adults emerging relative to a control diet lacking GNA, whereas differences were not observed among the three treatment levels. Both larvae and pupae in the control were 8-25% larger than those in the GNA treatments, but differences were not observed between larvae in the GNA treatments. Furthermore, presence of GNA did not affect larval and pupal developmental periods, longevities, and fecundities compared with the control. Mexican rice borer life history parameters, such as net reproductive rate and intrinsic rate of increase, were substantially reduced by the presence of GNA in the diet, but differences were not evident among the three GNA treatment levels.

Resistance to the Mexican rice borer (Lepidoptera: Crambidae) among Louisiana and Texas sugarcane cultivars.

A 2-yr study to evaluate Louisiana and Texas sugarcane, Saccharum spp., cultivars for resistance to the Mexican rice borer, Eoreuma loftini (Dyar) was conducted in two locations in Texas, chosen for having different infestation levels. Criteria for assessment of resistance included percentage of bored internodes and adult emergence holes, the latter used to determine the relative impact of each cultivar on the potential areawide buildup or reduction of adult E. loftini populations. A recently released cultivar, HoCP 85-845, seemed to lose a portion of its resistance under heavy E. loftini infestation pressure, suggesting its value only in moderate-to-low infestation conditions. Cultivar CP 70-321 was the most resistant. Results indicated that cultivar LCP 85-384 was significantly (P < 0.05) more susceptible than NCo 310, traditionally the most susceptible cultivar commercially produced in Texas. In 2001, LCP 85-384, which now represents 85% of the production area in Louisiana, had the greatest moth production per hectare (17,052 +/- 3,956) under the lower infestation pressure, significantly (P < 0.05) higher than HoCP 85-845 (3,038 +/- 2,353). In a portion of the test at the high-infestation location, high levels of sodium and magnesium salt stress (15-30-cm soil depth) were associated with higher E. loftini damage in all cultivars except HoCP 91-555 and CP 70-321.

Natural enemies of the maize cob borer, Mussidia nigrivenella (Lepidoptera: Pyralidae) in Benin, West Africa.

Mussidia nigrivenella Ragonot is a pest of maize cobs in West Africa. It significantly reduces maize yields and grain quality, with quantitative losses of 2-25%at harvest, and up to 10-15% indirect losses due to an increase in storage pest infestation levels. Infestation by M. nigrivenella also significantly increased the susceptibility of maize to Aspergillus flavus infection and subsequent aflatoxin contamination. Surveys conducted in different agro-ecological zones of Benin on cultivated and wild host plants during 1994-1997 revealed one egg parasitoid, three larval parasitoids and one pupal parasitoid attacking M. nigrivenella. Egg parasitism was scarce on all host plants sampled and in all four agro-ecological zones. Parasitism by larval and pupal parasitoids was usually less than 10%, and varied with host plant species. Both larval and pupal parasitoids were rare or absent in cultivated maize fields. The solitary chalcidid pupal parasitoid, Antrocephalus crassipes Masi, was the predominant species, contributing approximately 53% of the observed mortality. Logistic regression analysis indicated that this parasitoid was more prevalent on fruits of Gardenia spp. (Rubiaceae) than on the other host plant species including maize used by M. nigrivenella, and was most abundant between February and September. The differences in parasitoid diversity and parasitism between Benin and other regions suggest that there are opportunities for biological control through introduction of exotic parasitoids or using the 'new association' approach, which uses natural enemies of closely related host species that occupy similar ecological niches to the target pest.

Evaluation of lectin-expressing transgenic sugarcane against stalkborers (Lepidoptera: Pyralidae): effects on life history parameters.

The impact of snowdrop lectin (Galanthus nivalis agglutinin, GNA) expressed in transgenic sugarcane on life history parameters of Mexican rice borer [Eoreuma loftini (Dyar)] and sugarcane borer [Diatraea saccharalis (F.)] (both Lepidoptera: Pyralidae) was evaluated. In the laboratory, lyophilized sugarcane leaf sheath tissue was incorporated in a meridic diet resulting in a GNA concentration of 0.47% of total protein, and used for insect bioassays over two successive generations. Deleterious effects of GNA were not observed on survival, weight, and developmental periods of larvae and pupae, nor on adult fecundity and egg viability of D. saccharalis. Moreover, in the first generation, addition of transgenic sugarcane tissue to the diet enhanced larval growth in D. saccharalis resulting in higher larval and pupal weight compared with diet with nontransgenic sugarcane, but this effect was not observed in the second generation. In contrast, larval survival, percent adult emergence, and female fecundity of E. loftini were significantly reduced when fed transgenic sugarcane diet compared with nontransgenic sugarcane diet. In addition, a substantial reduction of female pupal weight of E. loftini was observed in the second generation. For both species, the only consistent effect of GNA in both generations was a reduction in adult female longevity. Life table parameters showed that GNA at the level found in the transgenic diet negatively affected development and reproduction of E. loftini, whereas it had a nil to positive effect on development and reproduction of D. saccharalis.

Monitoring and modeling of field infestation and damage by the maize ear borer Mussidia nigrivenella (Lepidoptera: Pyralidae) in Benin, West Africa.

In many countries in West Africa, the pyralid ear borer Mussidia nigrivenella Ragonot occasionally causes severe damage to pre- and postharvest maize. Between 1992 and 1995, the distribution of and damage caused by M. nigrivenella were studied in Benin using survey data and an on-station field experiment. The borer was distributed across the whole country, and at maturity an average 25% of the ears sampled in maize fields were infested. Damage levels varied with agro-ecological zones and were highest in the Guinea Savannas. However, borer-related yield losses were comparatively low. Three applications of cypermethrin over the growing season did not provide sufficient control in the on-station field experiment. A model was developed to estimate maize losses caused by M. nigrivenella, using the percentage of infested ears, which explained 93% of the variance. Extrapolation of field data indicated a 25% yield loss once a 100% infestation of maize ears was reached. For surveys in maize fields the model is a valid tool for a rapid assessment of crop losses caused by M. nigrivenella.

The influence of storage practices on aflatoxin contamination in maize in four agroecological zones of Benin, west Africa.

Aflatoxin level in 300 farmers' stores in four agro-ecological zones in Benin, a west African coastal country, were determined over a period of 2 years. At sampling a questionnaire was used to evaluate maize storage practices. Farmers were asked what storage structure they used, their storage form, storage period, pest problems in storage and what was done against them. Beninese farmers often changed their storage structures during the storage period, transfering the maize from a drying or temporary store to a more durable one. Most of the farmers complained about insects damaging stored maize. Often, storage or cotton insecticides were utilized against these pests. Regression analysis identified those factors that were associated with increased or reduced aflatoxin.Maize samples in the southern Guinea and Sudan savannas were associated with higher aflatoxin levels and the forest/savanna mosaic was related to lower toxin levels. Factors associated with higher aflatoxin were: storage for 3-5 months, insect damage and use of Khaya senegalensis-bark or other local plants as storage protectants. Depending on the agroecological zone, storage structures that had a higher risk of aflatoxin development were the "Ago", the "Secco", the "Zingo" or storing under or on top of the roof of the house. Lower aflatoxin levels were related to the use of storage or cotton insecticides, mechanical means or smoke to protect against pests or cleaning of stores before loading them with the new harvest. Fewer aflatoxins were found when maize was stored in the "Ago" made from bamboo or when bags were used as secondary storage containers.

Aspergillus flavus Infection and Aflatoxin Contamination of Preharvest Maize in Benin.

Eighty and sixty maize fields were sampled in 1994 and 1995, respectively, to monitor Aspergillus infection and aflatoxin contamination of preharvest maize in Benin. Three Aspergillus species were isolated from different agroecological zones, with A. flavus being the most prevalent. The countrywide mean percentage of kernel infection was about 20% in both years. Aflatoxin was extracted from maize in at least 30% of the fields sampled. Toxin concentrations exhibited a distinct zonal variation, with relatively high levels in the Guinea Savanna. There was a trend toward higher rate of aflatoxin accumulation per percentage A. flavus infection from the south to the north. Damage by the ear borer, Mussidia nigrivenella, increased aflatoxin accumulation in maize. Hence, the geographic pattern observed in the occurrence of A. flavus and aflatoxin may be related to the incidence of M. nigrivenella.