SARS-CoV-2 infection impacts carbon metabolism and depends on glutamine for replication in Syrian hamster astrocytes.

COVID-19 causes more than million deaths worldwide. Although much is understood about the immunopathogenesis of the lung disease, a lot remains to be known on the neurological impact of COVID-19. Here, we evaluated immunometabolic changes using astrocytes in vitro and dissected brain areas of SARS-CoV-2 infected Syrian hamsters. We show that SARS-CoV-2 alters proteins of carbon metabolism, glycolysis, and synaptic transmission, many of which are altered in neurological diseases. Real-time respirometry evidenced hyperactivation of glycolysis, further confirmed by metabolomics, with intense consumption of glucose, pyruvate, glutamine, and alpha ketoglutarate. Consistent with glutamine reduction, the blockade of glutaminolysis impaired viral replication and inflammatory response in vitro. SARS-CoV-2 was detected in vivo in hippocampus, cortex, and olfactory bulb of intranasally infected animals. Our data evidence an imbalance in important metabolic molecules and neurotransmitters in infected astrocytes. We suggest this may correlate with the neurological impairment observed during COVID-19, as memory loss, confusion, and cognitive impairment.

Alteration of hippocampal Egr3, GABA A receptors, Il-1ß, Il6 and Ccl3 expression in audiogenic tremor mice after seizure.

Neuroinflammation plays a protective role in the brain; however, in neurological diseases such as epilepsy, overactivated neuroinflammation, along with overexpression of inflammatory mediators, can cause neuronal tissue damage, which can trigger seizures due to loss of ionic or neurotransmitter homeostasis. Therefore, we aimed to evaluate mRNA expression levels of proinflammatory cytokines, early growth response factor 3 (Egr3), and GABA A receptors in the hippocampus of naive audiogenic mutant tremor mice, and stimulated tremor mice after a seizure. Gene expression of Il-1ß, Il-6, Tnf-α, Ccl2, Ccl3, Egr3, Gabra1, and Gabra4 from hippocampal samples of naive and stimulated tremor mice were measured by quantitative reverse transcription polymerase chain reaction (qRT-PCR). Relative to resistant mice, Ccl3 gene expression was increased and Il6 was decreased in the hippocampus of naïve tremor mice. Thirty minutes after a seizure, Ccl3 and Il-1ß mRNA expression were decreased (p < 0.0001; p = 0.0034, respectively) while Il6 was increased (p = 0.0052) in stimulated tremor mice, relative to naïve animals. In addition, Egr3, Gabra1, and Gabra4 mRNA expression was decreased in the hippocampus of naive tremor mice, relative to resistant mice, which increased 30 minutes after a seizure (p = 0.0496; p = 0.0447, and p = 0.0011, respectively), relative to naïve animals. In conclusion, overexpression of Ccl3 in the hippocampus of naive tremor mice, followed by downregulation soon after seizure in stimulated tremor mice, could be involved in changes in the blood-brain barrier (BBB) permeability in epilepsy. Il-1ß may be involved in hippocampal downregulation of GABA A receptors of naive tremor mice, characterizing an important mechanism in audiogenic seizures triggering. Hippocampal alterations of proinflammatory cytokines, Egr3, and GABA A receptors in tremor mice reinforce them as an alternative tool to modeling temporal lobe epilepsy.

Behavioral and neurochemical characterization of the spontaneous mutation tremor, a new mouse model of audiogenic seizures.

The tremor mutant phenotype results from an autosomal recessive spontaneous mutation arisen in a Swiss-Webster mouse colony. The mutant mice displayed normal development until three weeks of age when they began to present motor impairment comprised by whole body tremor, ataxia, and decreased exploratory behavior. These features increased in severity with aging suggesting a neurodegenerative profile. In parallel, they showed audiogenic generalized clonic seizures. Results from genetic mapping identified the mutation tremor on chromosome 14, in an interval of 5 cM between D14Mit37 (33.21 cM) and D14Mit115 (38.21 cM), making Early Growth Response 3 (Egr3) the main candidate gene. Comparing with wild type (WT) mice, the tremor mice showed higher hippocampal gene expression of Egr3 and Gabra1 and increased concentrations of noradrenalin (NOR; p = .0012), serotonin (5HT; p = .0083), 5-hydroxyindoleacetic acid (5-HIAA; p = .0032), γ-amino butyric acid (GABA; p = .0123), glutamate (p = .0217) and aspartate (p = .0124). In opposition, the content of glycine (p = .0168) and the vanillylmandelic acid (VMA)/NOR ratio (p = .032) were decreased. Regarding to dopaminergic system, neither dopamine (DA) and 3,4-dihydroxyphenylacetic acid (DOPAC) contents nor the turnover rate of DA showed statistically significant differences between WT and mutant mice. Data demonstrated that audiogenic seizures of tremor mice are associated with progressive motor impairment as well as to hippocampal alterations of the Egr3 and Gabra1 gene expression and amino acid and monoamine content. In addition, the tremor mice could be useful for study of neurotransmission pathways as modulators of epilepsy and the pathogenesis of epilepsies occurring with generalized clonic seizures.

Impacts of seven insecticides on Cotesia flavipes (Cameron) (Hymenoptera: Braconidae).

The endoparasitoid wasp Cotesia flavipes (Cameron) (Hymenoptera: Braconidae) is inundatively released in Brazilian sugarcane plantations to control the sugarcane borers Diatraea saccharalis (Fabricius) and Diatraea flavipennella (Box) (Lepidoptera: Crambidae). In conjunction with these releases, several synthetic insecticides are used to control the neonate larvae of these pests. We assessed the lethal and transgenerational sublethal effects of seven of these insecticides on C. flavipes. Leaf discs were sprayed at the highest field concentrations of chlorantraniliprole, lambda-cyhalothrin + chlorantraniliprole, chlorfluazuron, triflumuron, lambda-cyhalothrin + thiamethoxam, tebufenozide, and novaluron. Distilled water was used as a negative control. Newly emerged females (24 h old) were placed in Petri dishes containing the treated leaves, and the lethal and transgenerational sublethal effects were assessed for the next two generations. Lambda-cyhalothrin + chlorantraniliprole and lambda-cyhalothrin + thiamethoxam caused 100% mortality of the parasitoid and were highly persistent, causing more than 30% mortality at 30 days after spraying. Chlorantraniliprole, chlorfluazuron, novaluron, and triflumuron did not cause significant mortality compared to the negative control, but did have transgenerational sublethal effects. The length of the tibia of the right posterior leg, used as a growth measurement, was reduced in the progeny (F1 generation) of exposed female parasitoids. In addition, chlorantraniliprole increased and chlorfluazuron reduced the proportion of females in the F1 generation, whereas novaluron reduced the proportion of females in the F2 generation. Overall, only tebufenozide was considered harmless to C. flavipes. The results of this study suggest that lambda-cyhalothrin + chlorantraniliprole and lambda-cyhalothrin + thiamethoxam are harmful to C. flavipes, although field studies are needed to obtain results for actual sugarcane crops.

Oral acute toxicity and impact of neonicotinoids on Apis mellifera L. and Scaptotrigona postica Latreille (Hymenoptera: Apidae).

Wild and managed bees are essential for crop pollination and food production. However, the widespread use of insecticides such as neonicotinoids may affect the survival, development, behavior, and maintenance of bee colonies. Therefore, in this study we evaluated the impacts of three neonicotinoid insecticides on the survival and walking abilities of the Africanized honeybee A. mellifera and stingless bee S. postica. A. mellifera was more susceptible than S. postica to all neonicotinoids tested. The median lethal concentrations LC50 values estimated for acetamiprid, imidacloprid, and thiacloprid were 189.62, 22.78, and 142.31 ng µL-1 of diet for A. mellifera, and 475.94, 89.11, and 218.21 ng µL-1 of diet for S. postica, respectively. All tested neonicotinoids affected the speed, distance traveled, duration and frequency of resting, and continuous mobility of both bee species. The results showed that in spite of the different susceptibility to compounds with cyano and nitro radicals, the behavioral variables showed different levels of commitment according to the molecule insecticide and bee species. These results contribute not only to the understanding of the effects of neonicotinoid insecticides on A. mellifera and S. postica, but also to help in the development of protocols that aim to reduce the impact of these insecticides in Neotropical environments.

The Asian Citrus Psyllid Host Murraya koenigii Is Immune to Citrus Huanglongbing Pathogen 'Candidatus Liberibacter asiaticus'.

The Asian citrus psyllid (ACP) Diaphorina citri, vector of 'Candidatus Liberibacter asiaticus' (CLas), the putative causal agent of citrus Huanglongbing (HLB), is controlled by application of insecticides, which, although effective, has resulted in serious biological imbalances. New management tools are needed, and the technique known as "trap crop" has been attracting attention. A potential plant for use as a trap crop in the management of the ACP is Murraya koenigii (curry leaf). However, for this plant to be used in the field, it needs to be attractive for the vector and must not harbor CLas. To verify the potential of curry leaf as trap crop for the management of HLB, we investigated the ability of D. citri to transmit CLas to M. koenigii, and to other test plants, including M. paniculata (orange jasmine) and cultivar Valencia sweet-orange seedlings. For the tests, the insects were reared on a symptomatic CLas-infected plant and allowed to feed on the three test plant species. The overall maximum transmission rate for the citrus seedlings was 83.3%, and for orange jasmine was 33.3%. Successful transmission of CLas by ACP to the curry-leaf seedlings was not observed, and it was treated as immune to CLas. Supported by the previous results that M. koenigii is attractive for ACP, these results indicate that curry leaf is an excellent candidate for use as a trap crop, to improve the management of the insect vector and consequently of HLB.

Sublethal effects of insecticides used in soybean on the parasitoid Trichogramma pretiosum.

To control crop pests, parasitoid wasps of the genus Trichogramma are one alternative to the use of insecticides. Since a wide variety of agrochemicals may be applied to the same crops, it is essential to assess the selectivity of insecticides used for pest control on Trichogramma pretiosum. Information on which insecticides are less harmful to T. pretiosum can improve biological control using this insect, an important tactic in IPM programs for field crops. This study aimed to determine the effects of insecticides on the pupal stage and on the parasitism capacity of T. pretiosum. Lambda-cyhalothrin + thiamethoxam were slightly harmful and chlorpyriphos was moderately harmful to the pupal stage, while acephate, chlorfenapyr and flubendiamide, although considered innocuous, affected the succeeding generations of wasps, with low emergence of F1. Chlorfenapyr, chlorpyriphos and lambda-cyhalothrin + thiamethoxam reduced the parasitism, and acephate had a deleterious effect on the generation that contacted the insecticide residue. For an effective IPM program, it is important to apply selective insecticides. Further studies are needed to determine the selectivity of these insecticides under field conditions.

Spraying pyrethroid and neonicotinoid insecticides can induce outbreaks of Panonychus citri (Trombidiformes: Tetranychidae) in citrus groves.

Panonychus citri (McGregor) (Trombidiformes: Tetranychidae) is one of the main defoliator mites in citrus groves. In Brazil, P. citri was formerly considered a secondary pest, but in recent years, the population levels of this mite have increased in many groves, requiring additional acaricide applications for its control. The population growth of P. citri is associated with the increase in the number of applications of pyrethroid and neonicotinoid insecticides to control insect citrus pests. This study assessed the effects of pyrethroid (deltamethrin, esfenvalerate, λ-cyhalothrin) and neonicotinoid (imidacloprid and thiamethoxam) insecticides on biological, behavioral and demographic parameters of P. citri. None of the insecticides tested affected larval hatching, but deltamethrin, esfenvalerate, and λ-cyhalothrin reduced the survival of larvae and protonymphs. Deltamethrin, esfenvalerate, and λ-cyhalothrin induced a significant increase in the fecundity of surviving females. Pyrethroids also caused repellency and changed feeding and oviposition preferences. In contrast, imidacloprid and thiamethoxam did not affect the survival of immature stages, but imidacloprid significantly increased fecundity. Imidacloprid and thiamethoxam did not cause any repellent effect or changes in the feeding and oviposition preferences. Based on the demographic parameters, deltamethrin, λ-cyhalothrin, and imidacloprid increased Ro, r, and λ, whereas esfenvalerate and thiamethoxam were similar to the control. Therefore, the use of these insecticides (especially deltamethrin, λ-cyhalothrin, and imidacloprid) requires caution, such as avoiding repeated use of these compounds during the periods of P. citri incidence in citrus groves.

Sublethal effects of pyrethroid and neonicotinoid insecticides on Iphiseiodes zuluagai Denmark and Muma (Mesostigmata: Phytoseiidae).

The predator mite Iphiseiodes zuluagai Denmark & Muma is an important biological-control agent of mite pests, and it is one of the most common species found in citrus orchards. This study assessed, under laboratory conditions, the toxicity and duration of the harmful effects of five insecticides, the three pyrethroids deltamethrin, esfenvalerate and lambda-cyhalothrin, and the two neonicotinoids imidacloprid and thiamethoxam on I. zuluagai. Furthermore, we estimated the life-table parameters of the predator. Our results showed that deltamethrin and lambda-cyhalothrin caused higher mortality of larvae and adults than imidacloprid and thiamethoxam. In contrast, esfenvalerate provided larval mortality similar to imidacloprid and thiamethoxam, but it did not cause significant adult mortality of the predator. Mites that developed on pyrethroid residues showed lower survival of the immature stages, fecundity, and longevity compared to neonicotinoid residues and the control treatment. The estimated life-table parameters indicated that deltamethrin, lambda-cyhalothrin and esfenvalerate caused greater reduction in R o and r of I. zuluagai compared with imidacloprid and thiamethoxam, which were similar to the control treatment. Besides the impacts on biological and population parameters, the duration of the harmful activity of pyrethroid insecticides was longer than the neonicotinoids. Therefore, the use of pyrethroid insecticides to control pest insects may involve serious implications for integrated pest-management programs that aim to exploit the biological control by I. zuluagai in citrus orchards.

Susceptibility of Euseius concordis (Mesostigmata: Phytoseiidae) to pesticides used in citrus production systems.

Euseius concordis (Chant) is an important predatory mite found in citrus orchards. The toxicity of 19 pesticides used in citrus orchards on biological and population parameters of this mite was assessed. Our results indicated that formetanate hydrochloride, dimethoate and phosmet were highly harmful (100% mortality) to E. concordis. Carbosulfan, diflubenzuron, fenpropathrin, gamma-cyhalothrin, imidacloprid, lambda-cyhalothrin, lambda-cyhalothrin + thiamethoxam, mineral and vegetable oils, spinosad and thiamethoxam reduced the female's survival and/or fecundity, and were moderately harmful to E. concordis. Besides the acute toxicity, carbosulfan and formetanate hydrochloride were highly persistent [>30 days after spraying (DAS)]; dimethoate was moderately persistent (16-30 DAS); spinosad, gamma-cyhalothrin, lambda-cyhalothrin and lambda-cyhalothrin + thiamethoxam were slightly persistent (5-15 DAS); and the other pesticides were considered to be short-lived (<5 DAS). All compounds except lambda-cyhalothrin and thiamethoxam increased the pre-oviposition period in the female offspring. Carbosulfan, deltamethrin, diflubenzuron, etofenprox, fenpropathrin, gamma-cyhalothrin, mineral and vegetable oils, pyriproxyfen and tebufenozide reduced offspring fecundity, whereas thiamethoxam increased the fecundity. Mineral and vegetable oils reduced female longevity of the predator mite. Regarding population effects, imidacloprid, lambda-cyhalothrin, lambda-cyhalothrin + thiamethoxam and thiamethoxam led to an increase in net reproductive rate (R o ), intrinsic rate of increase (r), and finite rate of increase (λ) of E. concordis. Diflubenzuron, etofenprox, and mineral and vegetable oils reduced R o , r and λ. All pesticides except beta-cypermethrin, fenpropathrin and imidacloprid reduced the mean generation time (T) of the predator. Therefore, semi-field and field studies are needed to assess the compatibility of these compounds with E. concordis before adoption in IPM programs.

Impact of five insecticides used to control citrus pests on the parasitoid Ageniaspis citricola Longvinovskaya (Hymenoptera: Encyrtidae).

The parasitoid Ageniaspis citricola Longvnovskaya is a main biological control agent of the citrus leafminer Phyllocnistis citrella Stainton, an insect pest that causes considerable damage to citrus worldwide. However, the use of pesticides to control arthropod pests can reduce the effectiveness of parasitoids and disrupt integrated pest management in citrus groves. This study evaluated the impact on A. citricola of five insecticides that are used to control arthropod pests in citrus. Our results indicated that imidacloprid, chlorpyrifos, bifenthrin and ß-cyfluthrin were harmful (mortality >89 %) to A. citricola adults; whereas abamectin did not cause significant mortality and was considered harmless to the parasitoid. In addition to causing high mortality, imidacloprid and bifenthrin were considered moderately persistent, because they caused <25 % mortality to 17 and 24 days after spraying (DAS), respectively. Chlorpyrifos and ß-cyfluthrin were considered slightly persistent (mortality <25 %, 7 DAS). Although abamectin was considered harmless to A. citricola adults, had a short life (mortality <25 %, 3 DAS), and did not significantly affect the parasitism rate, the number and viability of pupae, or the longevity of A. citricola, this insecticide significantly reduced the proportion of females in the progeny compared to the control treatment. Therefore, semi-field and field studies that consider demographic parameters are needed to evaluate the impacts of these insecticides on the A. citricola parasitoid.

Impact of insect growth regulators on the predator Ceraeochrysa cincta (Schneider) (Neuroptera: Chrysopidae).

The generalist predator Ceraeochrysa cincta (Schneider) (Neuroptera: Chrysopidae) is an important biological control agent of several arthropod pests in different agroecosystems. This study assessed the lethal and sublethal effects of six insect growth regulators sprayed on first-instar larvae of C. cincta. Lufenuron and diflubenzuron were highly harmful to first-instar larvae of C. cincta, causing 100 % of mortality before they reached the second instar. Buprofezin caused ~25 % mortality of the larvae and considerably reduced the fecundity and longevity of the insects, but substantially increased the proportion of females in the surviving population of C. cincta. Methoxyfenozide and tebufenozide did not affect the duration and survival of the immature stages, but methoxyfenozide significantly reduced the fecundity and longevity of the insects. Pyriproxyfen reduced the survival of the larval stage by 19.5 %, but did not affect the development, survival and reproduction of the surviving individuals. Based on reduction coefficient, the insecticides diflubenzuron and lufenuron were considered harmful to C. cincta, whereas buprofezin and methoxyfenozide were slightly harmful and tebufenozide and pyriproxyfen were harmless. The estimation of life-table parameters indicated that buprofezin and methoxyfenozide significantly reduced the R o , r and λ of C. cincta, whereas pyriproxyfen and tebufenozide caused no adverse effect on population parameters, indicating that these insecticides could be suitable for use in pest management programs towards the conservation and population increase of the predator in agroecosystems. However, more studies should be conducted to evaluate the compatibility of these insecticides with the predator C. cincta under semi-field and field conditions.

Are the Pupae and Eggs of the Lacewing Ceraeochrysa cubana (Neuroptera: Chrysopidae) Tolerant to Insecticides?

The tolerance of Ceraeochrysa cubana (Hagen) pupae and eggs to 11 insecticides was evaluated under laboratory conditions, based on lethal and sublethal effects. Eggs at three ages (≤24-h-old, 48- to 72-h-old, and 96- to 120-h-old) and pupae at ≤48-h-old were used. All the insecticides were considered harmless when applied at the pupal stage. Phosmet and pyriproxyfen insecticides were considered harmless to eggs irrespective of the age. Esfenvalerate was harmless to eggs at the ages of ≤24-h-old and 48- to 72-h-old. Imidacloprid SC and azadirachtin were harmless to eggs at ages of 48- to 72-h-old and 96- to 120-h-old, and thiamethoxam was only harmless to eggs at 96- to 120-h-old of age. In contrast, chlorpyrifos and malathion were harmful to eggs at the age of 96- to 120-h-old, and imidacloprid WG was slightly harmful to the three egg ages evaluated. Lambda-cyhalothrin + chlorantraniliprole and lambda-cyhalothrin + thiamethoxam were slightly and/or moderately harmful to all egg ages evaluated. Based on the life table parameters, the insecticides thiamethoxam, imidacloprid SC, phosmet, pyriproxyfen, and azadirachtin did not affect the net rate of reproduction (Ro) of C. cubana. Lambda-cyhalothrin + chlorantraniliprole decreased the Ro and increased the population doubling time (Td) independently of the egg ages evaluated. Therefore, the insecticides pyriproxyfen and phosmet are compatible with eggs of the predator C. cubana, but other insecticides should be evaluated under field conditions to verify their effects on the predator.

Development and reproduction of Panonychus citri (Prostigmata: Tetranychidae) on different species and varieties of citrus plants.

The species and varieties of citrus plants that are currently grown can favor the population growth of the citrus red mite Panonychus citri (McGregor) (Prostigmata: Tetranychidae) and alter the pest management programs in citrus groves. In this study we evaluated, in the laboratory, the development and reproduction of P. citri and estimated its life table parameters when reared on four varieties of Citrus sinensis (L.) Osbeck (Valencia, Pera, Natal, and Hamlin), one variety of Citrus reticulata Blanco (Ponkan) and one variety of Citrus limon (L.) Burm. (Sicilian). The incubation period and egg viability were not affected by the host plant. However, the development and survival of the immature stage were significantly lower on Hamlin orange than on Valencia, Pera and Natal oranges, Ponkan mandarin and Sicilian lemon. The fecundity and oviposition period of females were lower on Hamlin orange than on the other hosts. Mites reared on Valencia orange and Sicilian lemon had a higher net reproductive rate (R 0 ), intrinsic growth rate (r) and finite rate of increase (λ), and a shorter interval between generations (T) than on Pera, Natal and Hamlin oranges and Ponkan mandarin. On the other hand, mites reared on Hamlin orange had the lowest R 0 , r and λ and the highest T among the hosts. Based on the results obtained we recommend that for Valencia orange and Sicilian lemon, the mite monitoring programs should be more intense to detect the initial infestation of pest, avoiding the damage in plants and the increase in production costs.

Comparative toxicity of an acetogenin-based extract and commercial pesticides against citrus red mite.

Acetogenins, a class of natural compounds produced by some Annonaceae species, are potent inhibitors of mitochondrial electron transport systems. Although the cellular respiration processes are an important biochemical site for the acaricidal action of compounds, few studies have been performed to assess the bioactivity of acetogenin-based biopesticides on spider mites, mainly against species that occur in orchards. Using residual contact bioassays, this study aimed to evaluate the bioactivity of an ethanolic extract from Annona mucosa seeds (ESAM) (Annonaceae) against the citrus red mite Panonychus citri (McGregor) (Acari: Tetranychidae), an important pest of the Brazilian citriculture. ESAM is a homemade biopesticide which was previously characterized by its high concentration of acetogenins. It caused both high mortality of P. citri females (LC50 = 7,295, 4,662, 3,463, and 2,608 mg l(-1), after 48, 72, 96, and 120 h of exposure, respectively) and significant oviposition deterrence (EC50 = 3.194,80 mg l(-1)). However, there was no effect on P. citri female fertility (hatching rate). In addition, the ESAM efficacy (in terms of its LC90) was compared with commercial acaricides/insecticides (at its recommended rate) of both natural [Anosom(®) 1 EC (annonin), Derisom(®) 2 EC (karanjin), and Azamax(®) 1.2 EC (azadirachtin + 3-tigloylazadirachtol)] and synthetic origin [Envidor(®) 24 SC (spirodiclofen)]. Based on all of the analyzed variables, the ESAM exhibited levels of activity superior to other botanical commercial acaricides and similar to spirodiclofen. Thus, our results indicate that ESAM may constitute a biorational acaricide for citrus red mite integrated pest management in Brazilian citrus orchards, particularly for local use.

Sublethal and Transgenerational Effects of Reduced-Risk Insecticides on Macrolophus basicornis (Hemiptera: Miridae).

Reduced-risk insecticides and mirid predators have been used to control Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) in tomato crops. However, even when causing low mortality to the beneficial insects, these products might cause side effects. This study investigated the sublethal and transgenerational effects of buprofezin, cyantraniliprole, and spiromesifen on Macrolophus basicornis (Stal) (Hemiptera: Miridae). After 72 h of exposure of third-instar nymphs and adults to residues on tomato leaves, adult couples were formed and kept in cages with residue-free tomato leaves. The leaves were changed every 48 h and the offspring were assessed in 6 different periods. Body size was assessed by measuring the hind-tibia length of adults (F0) from exposed nymphs and in three different offspring groups. None of the insecticide residues caused a reduction on offspring populations or affected the body size of adults in generation F0. Regardless, buprofezin and spiromesifen reduced the tibia length of adults (F1) from exposed nymphs assayed in the third mating period. Cyantraniliprole did not affect any parameter and could be recommended for control of B. tabaci in association with M. basicornis releases. This study may contribute to future field assays of the compatibility of these insecticides with M. basicornis.

Efficacy of Area-Wide Inoculum Reduction and Vector Control on Temporal Progress of Huanglongbing in Young Sweet Orange Plantings.

Huanglongbing (HLB), caused by 'Candidatus Liberibacter' spp. and transmitted by the Asian citrus psyllid Diaphorina citri (ACP), is an important threat to citrus industries worldwide, causing significant yield loss. The current recommended strategies to manage HLB are to eliminate HLB-symptomatic trees to reduce sources of bacterial inoculum and to apply insecticides to reduce psyllid vector populations. The objective of this study was to assess the effectiveness and the importance of both strategies applied within young citrus plots (local management), in different frequencies and combinations, on HLB temporal progress. Two factorial field experiments, E1 and E2, were initiated in a new plantation of sweet orange in an HLB epidemic region of Sao Paulo, Brazil, in October 2005 and May 2006, respectively. Local inoculum reduction (tree removal) intervals for E1 were every 4, 8, and 16 weeks and, for E2, every 2, 4, 12, and 26 weeks. Local vector control strategies for E1 were no control, program A (PA), and program B (PB); and, for E2, no control and program C (PC), as follows. Psyllids were controlled with two 56-day-interval soil or drench applications of systemic insecticides concurrently with the rainy season each year and, during the rest of the year, with insecticide sprays every 28 days for PA and every 14 days for PB and PC. Regional HLB management was present for E1 and absent for E2. The beginning of the HLB epidemic was delayed for 10 months in E1 compared with appearance of the first diseased tree in E2 but wasn't affected by different local strategies in either experiment. After 60 (E1) and 53 (E2) months, the HLB incidence and progress rates were not affected by different frequencies of local inoculum reduction in either experiment, and were different only in plots with and without local vector control in E2. In E1, the disease incidence was reduced by 90% and the disease progress rate by 50% in plots both with and without vector control. These reductions were explained by smaller psyllid populations and lower frequency of bacterialiferous psyllids in E1 compared with E2. Annual productivity increased over time in E1, as expected for young plantings, but remained stable or decreased in E2. These results confirm that immigration of bacterialiferous ACP vectors plays a critical role in HLB epidemics and suggest that area-wide inoculum reduction and ACP management strongly affect HLB control.

Is Integrated Management of Gymnandrosoma aurantianum Possible with Trichogramma atopovirilia and Novel Products Used in Citrus Orchards in Brazil?

In Brazil, the citrus fruit borer, Gymnandrosoma aurantianum Lima, 1927, is a serious pest in orange orchards, causing an annual loss of 80 million US dollars, and is managed with multiple insecticide applications, often 56 in a single season. On the other hand, the parasitoid wasp Trichogramma atopovirilia Oatman & Platner, 1983 has the potential for controlling G. aurantianum by attacking its eggs. Considering the intensive insecticide applications in citrus orchards in Brazil to control the large complex of pests, especially Diaphorina citri Kuwayama, 1908, evaluation of the harmful effects of insecticides on T. atopovirilia is important to maximize efficiency in managing G. aurantianum. Here, we tested the effects of new products used in citrus orchards (cyantraniliprole, cyantraniliprole + abamectin, abamectin, sulfoxaflor, spinetoram, flupyradifure, and Cordyceps fumosorosea (Wize) Kepler, B. Shrestha & Spatafora) on adults and pupae of T. atopovirilia. Of the insecticides tested, spinetoram caused the highest impacts on T. atopovirilia parasitism, longevity, emergence, and mortality. The other products caused more sublethal than lethal effects and were classified as 1 and/or 2 in the IOBC/WPRS classification. Abamectin, cyantraniliprole, cyantraniliprole + abamectin, and the entomopathogenic fungus C. fumosorosea were classified as short-lived. Except for spinetoram, these products were classified as selective. In this study, spinetoram was considered harmful to T. atopovirilia and, therefore, should be managed carefully in IPM programs combining this parasitoid. In order to safely use this insecticide, one should respect the interval of release of the parasitoid, which is 21 days after its spraying. The novel products tested, cyantraniliprole, cyantraniliprole + abamectin, abamectin, sulfoxaflor, and the entomopathogenic fungi C. fumosorosea were selective and non-persistent to T. atopovirilia. These products are possible replacements for non-selective insecticides to achieve higher control from both chemical and biological tools.

Effect of Systemic Insecticides Applied via Drench on the Mortality of Diaphorina citri on Curry Leaf.

Huanglongbing (HLB), the most serious disease in citriculture, is caused by the bacteria Candidatus Liberibacter spp., which is transmitted by the Asian citrus psyllid (ACP) Diaphorina citri. HLB is mainly controlled with insecticides, necessitating the development of alternative methods, e.g., the use of trap plants such as curry leaf Bergera koenigii, which is highly attractive to the ACP. We evaluated the effects of the main systemic insecticides used by citrus growers, applied via drench to adults of D. citri on the curry leaf tree. We tested the persistence of three pesticides: thiamethoxam, thiamethoxam + chlorantraniliprole, and imidacloprid in protected cultivation and the field condition at 7, 14, 28, 42, 56, 70, 98, and 154 days after the application. Different concentrations of insecticides containing the active ingredient thiamethoxam were tested on adults to determine the LC10 and LC50. Finally, we assessed the sublethal effects on the oviposition and development of D. citri. The insecticides controlled the adults for long periods. However, in the field experiment, from 42 days after application there was a decrease in mortality caused by pesticides applied via drench, while in the protected cultivation, mortality did not decline until the last day of evaluation. The median lethal concentration (LC50) for thiamethoxam was 0.031 g of active ingredient per plant, and for thiamethoxam in a mixture, the LC50 was 0.028 g a.i. per plant. In the experiment with sublethal doses, D. citri did not oviposit on the treated plants. Our findings suggest that the attract-and-kill system using the curry leaf tree and systemic insecticides is effective for the control of D. citri and contributes to the integrated management of HLB.

Impact of Bemisia tabaci MEAM1 (Hemiptera: Aleyrodidae) on Soybean Yield and Quality Under Field Conditions.

Bemisia tabaci MEAM1 (Hemiptera: Aleyrodidae) is a key insect pest in soybean fields in Brazil but data are lacking on the relationship between pest abundance and crop yield and quality. Controlled infestation studies were conducted on caged soybean plants in the field over a two year period at two sites in Brazil. Differences in temperature in the two years affected population growth of B. tabaci, reaching 413 nymphs per leaflet in the first year, and 179 the second year even when the average temperature was 3ºC higher. Higher temperatures promoted a shorter lifecycle and nearly one more generation. Yield was affected with losses up to 500 kg/ha in 2017/2018 and 1,147 kg/ha in 2018/2019. A decrease in the weight of a thousand grains of 18 and 33 g was observed in the first and second year, respectively. No significant differences were observed in grain germination, but estimated losses in protein content were up to 440 kg/ha at the highest infestation level. Pest density and yield data were used to estimate economic injury levels (EILs). EILs ranged from 2.5 to 25.67 nymphs per leaflet and 0.17-1.79 adults per leaflet over a range of control costs, soybean production values, and control efficacies. These results should provide data useful toward development of pest management decision making tools.