Regional susceptibilities to 12 insecticides of melon and cotton aphid,Aphis gossypii (Hemiptera: Aphididae) and a point mutation associated with imidacloprid resistance

The melon and cotton aphid Aphis gossypii Glover (Hemiptera; Aphididae) is one of the most serious pests worldwide. We surveyed insecticide susceptibility in A. gossypii field populations to 12 insecticides (6 neonicotinoids, 3 pyrethroids and 3 others) to examine resistance ratios. The levels of insecticide resistance were extremely high, especially to neonicotinoids, such as acetamiprid, clothianidin, thiacloprid and imidacloprid. To identify the neonicotinoid resistance mechanisms, we used an imidacloprid-resistant (IMI-R) strain as a model strain. IMI-R showed an extremely high resistance ratio and also cross-resistance to all the test neonicotinoids. However, there was little or no cross-resistance to the other insecticides, including sulfoxaflor. Synergist tests and enzyme activity assays suggested the absence of resistance mechanisms based on enhanced detoxification enzymes, such as cytochrome P450, esterase and glutathione S-transferase. One point mutation was found in the beta1 subunit loop D region of the nicotinic acetylcholine receptor (nAChR) of the IMI-R strain. This R81T point mutation was also found in field populations collected from 5 regions. Therefore, the R81T point mutation was identified as an important mechanism of imidacloprid resistance in A. gossypii.     

Functional Diversification of Oyster Big Defensins Generates Antimicrobial Specificity and Synergy against Members of the Microbiota

Big defensins are two-domain antimicrobial peptides (AMPs) that have highly diversified in mollusks. Cg-BigDefs are expressed by immune cells in the oyster Crassostrea gigas, and their expression is dampened during the Pacific Oyster Mortality Syndrome (POMS), which evolves toward fatal bacteremia. We evaluated whether Cg-BigDefs contribute to the control of oyster-associated microbial communities. Two Cg-BigDefs that are representative of molecular diversity within the peptide family, namely Cg-BigDef1 and Cg-BigDef5, were characterized by gene cloning and synthesized by solid-phase peptide synthesis and native chemical ligation. Synthetic peptides were tested for antibacterial activity against a collection of culturable bacteria belonging to the oyster microbiota, characterized by 16S sequencing and MALDI Biotyping. We first tested the potential of Cg-BigDefs to control the oyster microbiota by injecting synthetic Cg-BigDef1 into oyster tissues and analyzing microbiota dynamics over 24 h by 16S metabarcoding. Cg-BigDef1 induced a significant shift in oyster microbiota β-diversity after 6 h and 24 h, prompting us to investigate antimicrobial activities in vitro against members of the oyster microbiota. Both Cg-BigDef1 and Cg-BigDef5 were active at a high salt concentration (400 mM NaCl) and showed broad spectra of activity against bacteria associated with C. gigas pathologies. Antimicrobial specificity was observed for both molecules at an intra- and inter-genera level. Remarkably, antimicrobial spectra of Cg-BigDef1 and Cg-BigDef5 were complementary, and peptides acted synergistically. Overall, we found that primary sequence diversification of Cg-BigDefs has generated specificity and synergy and extended the spectrum of activity of this peptide family.     

Susceptibility levels of Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae) to minor classes of insecticides in Brazil

The tomato leafminer, Tuta absoluta (Meyrick), is one of the most important pests of tomato worldwide. The use of chemicals is still the major tactic of control against this pest in Brazil, where spray overuse leads to resistance, frequently associated with control failures in the field. In this study, a survey of the susceptibility to indoxacarb, metaflumizone, chlorfenapyr, cartap, and abamectin aimed to determine the resistance status of T. absoluta populations. Also, the major enzyme systems associated with metabolic resistance were assessed to infer variability. The LC₅₀ values varied among the populations for abamectin (0.54–3.38 mg a.i./L), cartap (93.1–589.8 mg a.i./L), chlorfenapyr (0.62–2.83 mg a.i./L), indoxacarb (0.86–2.89 mg a.i./L), and metaflumizone (0.35–7.44 mg a.i./L). Resistance ratios varied among populations being 6.2-, 6.4-, 4.6-, 3.3-, and 21.2-times for abamectin, cartap, chlorfenapyr, indoxacarb, and metaflumizone, respectively. Only the cartap confidence limits of the LC₈₀ bracketed the recommended label concentration for three populations (Anápolis, Guaraciaba do Norte, and Tianguá), suggesting control failures. No cross-resistance was observed between indoxacarb and metaflumizone, and natural variation may explain the variability of response to this latter insecticide. The activity of enzymes frequently associated with metabolism of insecticides significantly differed among populations, and glutathione S-transferases and cytochrome P450-dependent monooxygenases were variable among the populations of T. absoluta, while alpha and beta-esterases were very homogeneous. T. absoluta resistance to abamectin and cartap has not been critical in Brazil despite their long use and together with chlorfenapyr appear to be an interesting option of rotation with indoxacarb, for which there is no cross-resistance to metaflumizone.     

The dynamics of in vitro 17 β-estradiol secretion by isolated ovarian follicles of the rainbow trout (Oncorhynchus mykiss)

This study investigated the effects of human chorionic gonadotropin (hCG), 17-hydroxyprogesterone (17P) and testosterone (T) on the in vitro production of 17-estradiol (E₂) by isolated ovarian follicles of the rainbow trout (Oncorhynchus mykiss). 17P at 100 ng ml^-1, and hCG at 100 IU ml^-1 stimulated E₂ production relative to controls, whereas lower doses were ineffective. T was the most effective in stimulating E₂ production, followed by 17P and hCG respectively. The timecourse of E₂ production was investigated for both static culture, and incubations with media replacement, with follicles being exposed to hormone treatment for 30 min, 1 or 3 h, or constantly. E₂ production was observed after 30 min, 3 and 3-6 h in response to T, 17P and hCG respectively. Under static culture, E₂ levels reached maximal levels in 6 h. Longer incubations resulted in further metabolism of E₂ to E₂-glucuronide, which resulted in the blurring of treatment effects after 18 h. Incubations with media replacement resulted in higher E₂ production than in static culture. The results indicate that a 6 h incubation period is sufficient to produce significant increases in E₂ production in response to hCG, 17P and T, and that incubations longer than 12 h result in losses E₂ from the incubation media. These findings have implications for the validity of using static cultures to examine the effects of hormone treatment on the activity of steroid converting enzymes in vitro.     

Mechanism of action of sodium channel blocker insecticides (SCBIs) on insect sodium channels

Sodium channel blocker insecticides (SCBIs) are a relatively new class of insecticides, with a mechanism of action different from those of other classes of insecticides that target voltage-gated sodium channels. These compounds have no effect at hyperpolarized membrane potentials, but cause a voltage-dependent, nearly irreversible block as the membrane potential is depolarized. The mechanism of action of SCBIs is similar to that of local anesthetics (LAs), class I anticonvulsants and class I antiarrhythmics. In this article, we review the physiological actions of these compounds on the whole animal, the nervous system and sodium channels, and also present the results from recent studies that elucidate the receptor site of SCBIs.     

Acibenzolar-S-methyl Induces the Accumulation of Defense-related Enzymes in Apple and Protects from Fire Blight

Acibenzolar-S-methyl (Novartis) is a chemical inducer of systemic acquired resistance in several annual plants. The ability of this novel chemical to induce resistance was studied in a perennial plant (apple) affected by fire blight caused by Erwinia amylovora. Acibenzolar-S-methyl (100 and 200mg/l active ingredient) protected Golden Delicious seedlings, scions and trees from artificial infection when applied before inoculation. The protection of apple seedlings was similar to the protection obtained with the standard for fire blight control, plantomycin (100mg/l streptomycin sulfate), applied immediately before inoculation. The mean levels of control in scions in the greenhouse and in trees in orchards were approximately 69% and 50%, respectively. The protection of apple seedlings was constantly associated with the activation of two families of defense-related enzymes, peroxidases and -1,3-glucanases. Accumulation of both enzymes was induced locally in treated leaves and systemically, especially for -1,3-glucanases, in upper untreated leaves, and was sustained for at least 17 days. These results suggest that acibenzolar-S-methyl promotes induced systemic resistance in apple by increasing defense-related compounds. This chemical could provide a new approach of control of fire blight but its practical use needs further investigation.     

Modelling the progress of Asiatic citrus canker on Tahiti lime in relation to temperature and leaf wetness

The combined effect of temperature (15°C, 20°C, 25°C, 30°C, 35°C, 40°C and 42°C) and leaf wetness duration (0, 4, 8 12, 16, 20 and 24 h) on infection and development of Asiatic citrus canker (Xanthomonas citri subsp. citri) on Tahiti lime plant was examined in growth chambers. No disease developed at 42°C and zero hours of leaf wetness. Periods of leaf wetness as short as 4 h were sufficient for citrus canker infection. However, a longer leaf duration wetness (24 h) did not result in much increase in the incidence of citrus canker, but led to twice the number of lesions and four times the disease severity. Temperature was the greatest factor influencing disease development. At optimum temperatures (25–35°C), there was 100% disease incidence. Maximum disease development was observed at 30–35°C, with up to a 12-fold increase in lesion density, a 10-fold increase in lesion size and a 60-fold increase in disease severity.     

Efficiency of producing bioflocs with aquaculture waste by using poly-β-hydroxybutyric acid as a carbon source in suspended growth bioreactors

With additional organic carbon, fish waste can be used as a substrate to produce bioflocs, a protein source for aquaculture animals. In choosing a carbon source, one should consider convenience, cost and biodegradability. This study investigates the efficiency of poly-β-hydroxybutyric acid (PHB), a biologically degradable polymer, as a carbon source to produce bioflocs in suspended growth bioreactors (SGRs), PHB-SGRs, compared with glucose (GLU-SGRs). The C:N ratio in PHB-SGRs could be maintained around 15:1. The volatile suspended solids (VSS) yield was 2.94 ± 0.72 gVSS/g fish waste for PHB-SGRS and 4.90 ± 0.23 gVSS/g fish waste for GLU-SGRs. The recycling rate of nitrogen in aquaculture solid waste was 56 ± 2% and 87 ± 7% for the PHB-SGRs and Glu-SGRs. No significant differences were found in the bioflocs produced and in the crude protein content of the produced bioflocs between PHB-SGRs and GLU-SGRs. PHB-SGRs and GLU-SGRs could remove dissolved inorganic nitrogen from aquaculture wastewater, with average values of 11.82 ± 8.95 and 16.27 ± 3.95 mg/g TSS/d. Because the calculation of the added amount of carbon and the multiple additions of carbon was avoided, PHB is considered to be a good choice as an organic carbon source for this process, even though not all parameters used for assessment were better than those of GLU-SGRs.