The lived experiences and perceptions of middle-aged adults in Dominica who have survived severe storms-a qualitative exploration.

Introduction: The Eastern Caribbean island of Dominica has experienced diverse negative effects from the North Atlantic hurricane season, including deadly storms like Hurricane Maria in 2017. Vulnerability is increased by geographic location, small island developing state (SIDS) status, and ecosystem characteristics. A variety of negative health effects including stress and anxiety are caused by powerful storms. The perspectives of middle-aged (the "sandwich generation") survivors in this post-storm milieu are understudied. Methods: This phenomenological qualitative study describes the perceptions of middle-aged (35-55 years) Dominicans, purposively recruited with gatekeeper assistance from communities stratified according to four natural hazard vulnerability categories designated by the Climate Resilience Execution Agency for Dominica (CREAD), regarding their lived experiences in the context of severe storms. Data was collected between June and August 2022, using primarily Zoom-based semi-structured, individual interviews (12 of 13), guided by the principles of saturation and maximum variation. Verbatim interview transcripts were thematically analyzed with constant comparison using an ATLAS.ti-supported hybrid deductive-inductive coding frame. Reflexivity and contact summary sheets were used to minimize bias. Results: Ten women and three men from diverse CREAD vulnerability and sociodemographic backgrounds were recruited. Data condensation yielded three organizing themes: (i) "The diverse health effects of severe storms", (ii) "Response to and recovery from severe storms", and (iii) "Preparedness and precaution for severe storms". These themes encapsulated the health impacts of severe storms on Dominicans and elucidated the role of facilitating and barricading resilience factors. Discussion: Severe storms produced direct and indirect mental, social, and physical health impacts on middle-aged Dominicans, including anxiety and burnout. Participants used faith-based, tangible community-based support, and emotional mechanisms to cope with and demonstrate resilience. Better risk communication and early warning systems would improve population readiness. Persistent dispirited attitudes toward storm preparedness among some participants suggest the need for targeted methods to enhance community involvement in disaster planning, including traditional approaches like "coup-de-main" (self-help).

Trehalose 6-phosphate synthase gene rdtps1 contributes to thermal acclimation in Rhyzopertha dominica.

BACKGROUND: The lesser grain borer (Rhyzopertha dominica), a worldwide primary pest of stored grain, causes serious economic losses and threatens stored food safety. R. dominica can respond to changes in temperature, especially the adaptability to heat. In this study, transcriptome analysis of R. dominica exposed to different temperatures was performed to elucidate differences in gene expression and the underling molecular mechanism. RESULTS: Isoform-sequencing generated 17,721,200 raw reads and yielded 20,416 full-length transcripts. A total of 18,880 (92.48%) transcripts were annotated. We extracted RNA from R. dominica reared at 5 °C (cold stress), 15 °C (cold stress), 27 °C (ambient temperature) and 40 °C (heat stress) for RNA-seq. Compared to those of control insects reared at 27 °C, 119, 342, and 875 differentially expressed genes (DEGs) were identified at 5 °C, 15 °C, and 40 °C, respectively. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis revealed that pathways associated with "fatty acid metabolism", "fatty acid biosynthesis", "AMPK signaling pathway", "neuroactive ligand receptor interaction", and "longevity regulating pathway-multiple species" were significantly enriched. The functional annotation revealed that the genes encoding heat shock proteins (HSPs), fatty acid synthase (FAS), phospholipases (PLA), trehalose transporter (TPST), trehalose 6-phosphate synthase (TPS), and vitellogenin (Vg) were most likely involved in temperature regulation, which was also validated by RT-qPCR. Seven candidate genes (rdhsp1, rdfas1, rdpla1, rdtpst1, rdtps1, rdvg1, and rdP450) were silenced in the RNA interference (RNAi) assay. RNAi of each candidate gene suggested that inhibiting rdtps1 expression significantly decreased the trehalose level and survival rate of R. dominica at 40 °C. CONCLUSIONS: These results indicated that trehalose contributes to the high temperature resistance of R. dominica. Our study elucidates the molecular mechanisms underlying heat tolerance and provides a potential target for the pest management in R. dominica.

Stability evaluation of candidate reference genes for real-time qPCR normalization in Rhyzopertha dominica (Coleoptera: Bostrycidae).

Rhyzopertha dominica is a serious stored grain insect pest around the world. Real-time quantitative polymerase chain reaction (RT-qPCR) is a widely used experimental method in molecular biology for detecting the expression of target genes. As appropriate reference genes are essential for normalizing gene expression, the selection of suitable reference genes is the basis of RT-qPCR experiments. In this study, the expression profiles of 7 candidate reference genes of rps3, rps6, rps13, actin, gadph, tubulin, and 18S rRNA were analyzed under 4 different experimental conditions. The expression stability of candidate genes was evaluated using the ΔCt, GeNorm, BestKeeper, NormFinder, and RefFinder methods. The results revealed that different reference genes were suitable for various experiments. Specifically, rps3 and rps6 were appropriate for the developmental stages and all samples: 18S rRNA and rps13 for temperature-related experiments, actin and rps6 for sex-related experiments, and rps6 and gadph for starvation stress. Our results lay essential groundwork for the normalization of RT-qPCR analyses and contribute to genomic and gene functional research of R. dominica.

Regulation of Aquaporin Prip Expression and Its Physiological Function in Rhyzopertha dominica (Coleoptera: Bostrichidae).

Rhyzopertha dominica Prip (RdPrip) cDNA was cloned (GenBank accession no. OK318454), and the encoded 276-amino-acid protein indicated the typical aquaporin structure, including six transmembrane regions and two NPA motifs. The developmental and tissue profiles of RdPrip transcription were determined. RdPrip was highly transcribed in female adults, followed by larvae, pupae, and male adults. The transcriptional expression levels of RdPrip were significantly high in the ovary and hindgut (including cryptonephridial systems) compared with the foregut, testis, midgut, and Malpighian tubules. Knockdown of RdPrip in female adults did not decrease fecundity, but significantly decreased the hatching rate of eggs laid by the females. The results suggest that RdPrip functions in embryonic development, not in egg formation. In addition, the transcriptional expression level of RdPrip was lower in the spinosad-resistant strain than in the susceptible one, and the resistant strain produced fewer progeny than the susceptible strain did. These studies support the functional role of RdPrip in female reproduction. The absence of significant mortality reduction in the R. dominica exposed to spinosad after RdPrip RNAi suggests that other aquaporins that were not knocked down may exist for the excretion of metabolized pesticides.

Ozone reduces lifespan and alters gene expression profiles in Rhyzopertha dominica (Fabricius).

Rhyzopertha dominica is one of the most important stored grain pests that seriously damage rice and wheat. At present, the method of controlling stored grain pests mainly relies on insecticide fumigation. However, the excessive use of pesticides not only leaves pesticide residues, with harmful effects on human health and the environment, but also induces insect resistance. Ozone is a strong oxidant with the characteristics of easy decomposition and without residue. Although ozone has been widely used in the food industry in recent years, research on the control of stored grain pests is limited. In this research, we used ozone treatment to control R. dominica adults and explore the molecular mechanisms that affect them. Here, we found that ozone treatment on R. dominica adults could decrease life span and increase malondialdehyde (MDA) content, as well as reduce activity of total superoxide dismutase (SOD) and catalase (CAT). Using RNA-seq technology, we identified 641 genes that were differentially expressed between ozone-treated and control R. dominica adults [fold-change of ≥ 2 (q-value < 5%)]. When comparing ozone treatment with control R. dominica adults, 330 genes were significantly upregulated and 311 were downregulated. RT-qPCR confirmed that 11 genes were differentially expressed in ozone-treated and control R. dominica adults. These genes were involved in insect cuticle protein and antioxidant system. This research showed that ozone treatment could reduce the lifespan of R. dominica through antioxidant system. It is an environmentally benign method for the control of stored grain pests and has great development potential.

Formulation and characterisation of Azadirachta indica nanobiopesticides for ecofriendly control of wheat pest Tribolium castaneum and Rhyzopertha dominica.

This study aimed to formulate the green, sustainable, and ecofriendly nanobiopesticides of Azadirachta indica with enhanced pest control efficacy. Nanoprecipitation method was used for the development of nanobiopesticides. Optimisation was done by response surface methodology. Nanoformulations were characterised by zetasizer, scanning electron microscopy, energy dispersive x-ray spectroscopy, atomic force microscopy, and Fourier transform infrared spectroscopy. Pesticidal potential of nanosuspensions was evaluated by insecticide impregnated filter paper method. Optimised nanobiopesticide showed an average particle size of 275.8 ± 0.95 nm, polydispersity index (PDI) 0.351 ± 0.002, and zeta potential of -33 ± 0.90 mV. Nanobiopesticides exhibited significantly higher mortality rates of 86.81 ± 3.04 and 84.97 ± 2.83% against Tribolium castaneum and Ryzopertha dominica, respectively, as compared to their crude extract. Minor change in particle size from 275.8 ± 0.95 to 298.8 ± 1.00 nm and PDI from 0.351 ± 0.002 to 0.445 ± 0.02 were observed after 3 months of storage at 4 °C. Pesticidal efficacy of A. indica was significantly enhanced by the formulation of its nanobiopesticides.

Role of Lipids in Phosphine Resistant Stored-Grain Insect Pests Tribolium castaneum and Rhyzopertha dominica.

Insects rely on lipids as an energy source to perform various activities, such as growth, flight, diapause, and metamorphosis. This study evaluated the role of lipids in phosphine resistance by stored-grain insects. Phosphine resistant and susceptible strains of the two main stored-grain insects, Tribolium castaneum and Rhyzopertha dominica, were analyzed using liquid chromatography-mass spectroscopy (LC-MS) to determine their lipid contents. Phosphine resistant strains of both species had a higher amount of lipids than susceptible stains. Significant variance ratios between the resistant and susceptible strains of T. castaneum were observed for glycerolipids (1.13- to 53.10-fold) and phospholipids (1.05- to 20.00-fold). Significant variance ratios between the resistant and susceptible strains of R. dominica for glycerolipids were 1.04- to 31.50-fold and for phospholipids were 1.04- to 10.10-fold. Glycerolipids are reservoirs to face the long-term energy shortage. Phospholipids act as a barrier to isolate the cells from the surrounding environment and allow each cell to perform its specific function. Thus, lipids offer a consistent energy source for the resistant insect to survive under the stress of phosphine fumigation and provide a suitable environment to protect the mitochondria from phosphine. Hence, it was proposed through this study that the lipid content of phosphine-resistant and phosphine-susceptible strains of T. castaneum and R. dominica could play an important role in the resistance of phosphine.

Priorities for Bolstering Public Health Resilience in the Context of Climate Change in Dominica and Puerto Rico.

Caribbean small island developing states are highly exposed to climate change impacts. Incorporating weather and climate information into public health decisions can promote resilience to climate change's adverse health effects, but regionally it is not common practice. We implemented a project to enhance dialogue between climate and public health specialists in Puerto Rico and Dominica. First, we conducted environmental scans of public health vulnerability in the context of weather and climate for both islands. Then, we convened stakeholders to discuss the scan results and identify priorities for climate and health. A shared priority was increasing climate and health knowledge; thus, we developed several educational initiatives. In this viewpoint, we discuss our process for conducting environmental scans, building capacity and partnerships, and translating knowledge-to-action around climate and health.

Promising Insecticidal Efficiency of Essential Oils Isolated from Four Cultivated Eucalyptus Species in Iran against the Lesser Grain Borer, Rhyzopertha dominica (F.).

The lesser grain borer, Rhyzopertha dominica (F.), causes damage to stored grains resulting in both quantitative and qualitative losses. The use of synthetic fumigants in the management of stored-product pests resulted in undesirable side effects such as environmental contamination and threat to human and animal health. In this study, the lethal and sublethal effects of essential oils from four Eucalyptus species, E. microtheca, E. procera, E. spatulata, and E. torquata were studied against R. dominica adults. Gas chromatographic-mass spectral analysis of the essential oils was carried out, in which terpenes such as 1,8-cineole and globulol were abundant in essential oils. The pest was susceptible to the fumigation of the essential oils and, considering concentrations and exposure times (24, 48, and 72 h), had significant effects on the pest mortality. The total protein, glycogen, and lipid contents and digestive amylolytic and proteolytic activities of the adults treated with tested essential oils were reduced. The consumption index, relative consumption rate, and relative growth rate were also reduced in the treated adults. According to the insecticidal effects on the adults of R. dominica, the essential oils of E. microtheca, E. procera, E. spatulata, and E. torquata can be candidates for further investigations as grain protectant agents.

Chemical Profiles and Insecticidal Potential of Essential Oils Isolated from Four Thymus Species against Rhyzopertha dominica (F.).

Although chemical pesticides have been efficiently used to manage insect pest, their overuse has led to environmental contamination and threats to human health, enticing researchers to introduce eco-friendly and effective agents. In this study, the insecticidal effectiveness of essential oils isolated from Thymus species, including T. eriocalyx, T. kotschyanus, T. fallax, and T. vulgaris, was evaluated against the adults of Rhyzopertha dominica. The terpenes p-cymene, 1,8-cineole, linalool, α-terpineol, and carvacrol were the prominent compounds in the hydrodistilled essential oils. All essential oils produced significant fumigant at 24, 48, and 72-exposure times. The energy reserves protein by all essential oils, glycogen by T. kotschyanus and T. vulgaris, and lipid by T. fallax and T. vulgaris were significantly decreased compared to control. All essential oils except T. vulgaris affected the amylolytic and proteolytic activity of the pest. The pest increased the α- and ß-esterase enzyme activity in response to the essential oils. Nutritional indices of adults were also affected by essential oils, in which feeding deterrence index was calculated from 20.41% to 61.11%. Accordingly, based on lethal and extensive sub-lethal insecticidal activities, T. eriocalyx, T. kotschyanus, T. fallax, and T. vulgaris essential oils can be considered as efficient agents for R. dominica management.

Efficiency of a Novel Light-Emitting Diode (LED) Trap for Trapping Rhyzopertha dominica (Coleoptera: Bostrichidae) in Paddy Rice Storehouses.

The lesser grain borer Rhyzopertha dominica is the major pest of stored paddy rice globally, including in Taiwan. It has strong phototaxis and is good at flying, suitable for developing a light-trapping method to monitor and control it. In the present study, a wavelength of light-emitting diodes (LEDs), i.e., 373 nm, was determined to be the most efficient to trap R. dominica using a dodecagon maze. Accordingly, an LED trap, named the Taiwan Agricultural Research Institute-LED (TARI-LED) trap, was invented, which comprised LEDs of two distinct wavelengths (373 and 408 nm), a wavelength switch, a suction fan, and an insect collector. The trapping efficiency was assessed in a 4-m3 laboratory arena and two paddy rice storehouses. An initial assessment was performed in the laboratory arena and showed that the TARI-LED trap with 373-nm wavelength for R. dominica rapidly increased in the first 30 min, reaching the highest trapping rate (68.5%) after 3 h. In addition, no significant difference was observed between the suction fan turned on or off. The field tests showed that the 373-nm wavelength had the highest effectiveness for trapping R. dominica in the two paddy rice storehouses, and no significant difference was observed in the number of R. dominica trapped by the 373-nm TARI-LED trap or the CDC-UV light trap. In conclusion, our TARI-LED trap 373 nm exhibited high efficiency in trapping R. dominica in paddy rice storehouses. Moreover, a suction fan-free design should benefit long-term and safe use in paddy rice storehouses trapping R. dominica.

Returning to normal? 'Building back better' in the Dominican education system after Tropical Storm Erika and Hurricane Maria.

Child-centred disaster risk reduction aims to reduce child vulnerability and increase resilience to disasters. The 2015 Comprehensive School Safety Framework (CSSF) sought to decrease hazard risks to education. Between 2015 and 2017, Dominica was struck by Tropical Storm Erika and Hurricane Maria, which significantly affected the education system at the local and national scales. Since Maria, a couple of national initiatives (Safer Schools and Smart Schools) have been introduced to increase resilience and meet the CSSF's objectives. This paper assesses progress made through a qualitative analysis of interviews with 29 school leaders, government officials, and disaster risk reduction stakeholders. Implementation of the climate resilience programme in 2018 resulted in nationwide teacher training and production of school disaster plans. Limited successes have improved social resilience, but short-term implementation due to COVID-19 and a lack of a teacher knowledge base have presented challenges to the scheme's long-term sustainability and the implementation of the CSSF's goals.

The Genome of Rhyzopertha dominica (Fab.) (Coleoptera: Bostrichidae): Adaptation for Success.

The lesser grain borer, Rhyzopertha dominica (F.) (Coleoptera: Bostrichidae), is a major global pest of cereal grains. Infestations are difficult to control as larvae feed inside grain kernels, and many populations are resistant to both contact insecticides and fumigants. We sequenced the genome of R. dominica to identify genes responsible for important biological functions and develop more targeted and efficacious management strategies. The genome was assembled from long read sequencing and long-range scaffolding technologies. The genome assembly is 479.1 Mb, close to the predicted genome size of 480.4 Mb by flow cytometry. This assembly is among the most contiguous beetle assemblies published to date, with 139 scaffolds, an N50 of 53.6 Mb, and L50 of 4, indicating chromosome-scale scaffolds. Predicted genes from biologically relevant groups were manually annotated using transcriptome data from adults and different larval tissues to guide annotation. The expansion of carbohydrase and serine peptidase genes suggest that they combine to enable efficient digestion of cereal proteins. A reduction in the copy number of several detoxification gene families relative to other coleopterans may reflect the low selective pressure on these genes in an insect that spends most of its life feeding internally. Chemoreceptor genes contain elevated numbers of pseudogenes for odorant receptors that also may be related to the recent ontogenetic shift of R. dominica to a diet consisting primarily of stored grains. Analysis of repetitive sequences will further define the evolution of bostrichid beetles compared to other species. The data overall contribute significantly to coleopteran genetic research.

Variants in the Mitochondrial Genome Sequence of Rhyzopertha dominica (Fabricius) (Coleoptera: Bostrycidae).

The lesser grain borer, Rhyzopertha dominica, is a coleopteran pest of stored grains and is mainly controlled by phosphine fumigation, but the increase in phosphine-resistant populations threatens efficacy. Some phosphine-resistant insects have reduced respiration, and thus studying the mitochondrial genome may provide additional information regarding resistance. Genomic DNA from an inbred laboratory strain of R. dominica was extracted and sequenced with both short (Illumina) and long (Pacific Biosciences) read technologies for whole genome sequence assembly and annotation. Short read sequences were assembled and annotated by open software to identify mitochondrial sequences, and the assembled sequence was manually annotated and verified by long read sequences. The mitochondrial genome sequence for R. dominica had a total length of 15,724 bp and encoded 22 trna genes, 2 rRNA genes, 13 protein coding genes (7 nad subunits, 3 cox, 2 atp, and 1 cytB), flanked by a long control region. We compared our predicted mitochondrial genome to that of another from a R. dominica strain from Jingziguan (China). While there was mostly agreement between the two assemblies, key differences will be further examined to determine if mutations in populations are related to insecticide control pressure, mainly that of phosphine. Differences in sequence data, assembly, and annotation also may result in different genome interpretations.

Transcriptomic analysis of s-methoprene resistance in the lesser grain borer, Rhyzopertha dominica, and evaluation of piperonyl butoxide as a resistance breaker.

BACKGROUND: The lesser grain borer, Rhyzopertha dominica is a serious pest of stored grains. Fumigation and contact insecticides play a major role in managing this pest globally. While insects are developing genetic resistance to chemicals, hormonal analogues such as s-methoprene play a key role in reducing general pest pressure as well as managing pest populations that are resistant to fumigants and neurotoxic contact insecticides. However, resistance to s-methoprene has been reported in R. dominica with some reports showing a remarkable high resistance, questioning the use of this compound and other related analogues in grain protection. The current study attempts to identify possible molecular mechanisms that contribute in resistance to s-methoprene in R. dominica. RESULTS: Transcriptome analysis of resistant and susceptible strains of this pest species identified a set of differentially expressed genes related to cytochrome P450s, indicating their potential role in resistance to s-methoprene. Laboratory bioassays were performed with s-methoprene treated wheat grains in presence and absence of piperonyl butoxide (PBO), a cytochrome P450 inhibitor. The results indicate that PBO, when applied alone, at least at the concentration tested here, had no effect on R. dominica adult emergence, but has a clear synergistic effect to s-methoprene. The number of produced progeny decreased in presence of the inhibitor, especially in the resistant strain. In addition, we also identified CYP complement (CYPome) of R. dominica, annotated and analysed phylogenetically, to understand the evolutionary relationships with other species. CONCLUSIONS: The information generated in current study suggest that PBO can effectively be used to break resistance to s-methoprene in R. dominica.

Early warning systems and evacuation: rare and extreme versus frequent and small-scale tropical cyclones in the Philippines and Dominica.

Survey questionnaires were administered among populations affected by Super Typhoon Yolanda in the Philippines in 2013 and Hurricane Maria in Dominica in 2017 to test the efficacy of early warning systems in prompting residents to take appropriate action ahead of severe hazards. Both events were rare and extreme but occurred in locations that regularly experience less severe tropical cyclones. The research assessed if, how, and when residents received warnings, what instructions were given, and where and when people decided to seek safety. In both of the cases under review, residents were aware of the approaching storms, but critical information on their severity and potential impacts was either not received in time or not understood fully, resulting in low levels of evacuation and safety-seeking behaviour. This paper suggests that planning and public communication need to focus on the uncertainty surrounding the severity and multifaceted nature of tropical cyclones and accompanying hazards and their consequences.

Genetic diversity and ancestry of cacao (Theobroma cacao L.) in Dominica revealed by single nucleotide polymorphism markers.

Cacao (Theobroma cacao L.), an introduced tree crop in Dominica, is important for foreign exchange earnings from fine or flavour cocoa. The genetic structure of farmed cacao in Dominica was examined to identify varieties for conservation, breeding, and propagation to improve their cocoa industry. Cacao trees (156) from 73 sites over seven geographical regions were genotyped at 192 single nucleotide polymorphism (SNP) markers. Identity, regional differentiation, phylogenetic, multi-variate, ancestry, and core collection analyses were performed. Farmed cacao germplasm had moderate gene diversity (He = 0.320 ± 0.005) from generally unique trees, but cocoa growing regions were genetically similar. Synonymous matching (16.3%) showed that some clonal material was supplied to farmers. Cacao trees were mainly mixed from Amelonado, Criollo, Iquitos, Contamana, and Marañon ancestries, with predominantly Amelonado-Criollo hybrids. Criollo ancestry, linked to fine or flavour cocoa, was found at more than 30% in 28 unique trees. Forty-five trees, containing the SNP diversity of cacao in Dominica, are recommended as a core germplasm collection. This study identifies promising trees for improving cocoa quality; provides genetic evidence that community, regional, or country-wide pooling would not compromise the exclusive fine or flavour cocoa industry; and discusses other implications towards improving the Dominican cocoa industry.

Characterization of Chemical Components in Organs Emissions of Two Salvia dominica L. Populations from Jordan.

The current investigation reports the chemical composition of volatile organic compounds emitted from the different aerial organs of two populations of Salvia dominica L. from Jordan collected from Mediterranean and Irano-Turanian bio-geographical zones. Oxygenated monoterpenes dominated the emission profiles of most organs from the two populations but with variable qualitative and quantitative differences. Mediterranean samples contained higher content of oxygenated monoterpenes (stems: 88.37%, leaves 89.95%, pre-flowering buds 67.14%, fully opened flowers 79.43%, sepals 90.93% and petals 92.25%) as compared to those from Irano-Turanian origin (range 39.85% to 75.06%). trans-Sabinene hydrate dominated the emission profiles of all organs from Irano-Turanian zone (range 38.54% to 73.24%) in addition to the stem, sepal and petal samples from the Mediterranean zone (51.37% to 86.98%). The other organs from the Mediterranean zone were dominated by α-terpenyl acetate (27.66-54.87%). Cluster and Principle Component statistical analysis classified the two populations into two clusters based on their origin. The current study evidenced the different VOCs composition in the two populations, that was mainly related to climatic and environmental conditions and suggested the presence of two ecotypes of S. dominica L.in Jordan.

Preliminary Study on the Differences in Hydrocarbons Between Phosphine-Susceptible and -Resistant Strains of Rhyzopertha dominica (Fabricius) and Tribolium castaneum (Herbst) Using Direct Immersion Solid-Phase Microextraction Coupled with GC-MS.

Phosphine resistance is a worldwide issue threatening the grain industry. The cuticles of insects are covered with a layer of lipids, which protect insect bodies from the harmful effects of pesticides. The main components of the cuticular lipids are hydrocarbon compounds. In this research, phosphine-resistant and -susceptible strains of two main stored-grain insects, T. castaneum and R. dominica, were tested to determine the possible role of their cuticular hydrocarbons in phosphine resistance. Direct immersion solid-phase microextraction followed by gas chromatography-mass spectrometry (GC-MS) was applied to extract and analyze the cuticular hydrocarbons. The results showed significant differences between the resistant and susceptible strains regarding the cuticular hydrocarbons that were investigated. The resistant insects of both species contained higher amounts than the susceptible insects for the majority of the hydrocarbons, sixteen from cuticular extraction and nineteen from the homogenized body extraction for T. castaneum and eighteen from cuticular extraction and twenty-one from the homogenized body extraction for R. dominica. 3-methylnonacosane and 2-methylheptacosane had the highest significant difference between the susceptible and resistant strains of T. castaneum from the cuticle and the homogenized body, respectively. Unknown5 from the cuticle and 3-methylhentriacontane from the homogenized body recorded the highest significant differences in R. dominica. The higher hydrocarbon content is a key factor in eliminating phosphine from entering resistant insect bodies, acting as a barrier between insects and the surrounding phosphine environment.

New Method of Analysis of Lipids in Tribolium castaneum (Herbst) and Rhyzopertha dominica (Fabricius) Insects by Direct Immersion Solid-Phase Microextraction (DI-SPME) Coupled with GC-MS.

Lipids play an essential role in providing energy and other physiological functions for insects. Therefore, it is important to determine the composition of insect lipids from cuticular and internal tissues for a better understanding of insect biology and physiology. A novel non-derivatization method for the analysis of lipids including fatty acids, hydrocarbon waxes, sterols in Tribolium castaneum (Herbst) and Rhyzopertha dominica (Fabricius) was explored using the direct immersion solid-phase microextraction (DI-SPME) coupled with gas chromatography-mass spectrometry (GC-MS). Nine extraction solvents, acetonitrile, methanol, hexane, ethanol, chloroform, acetonitrile and ethanol (1:1 v/v), acetonitrile and water (1:1 v/v), ethanol and water (1:1 v/v) and acetonitrile and ethanol and water (2:2:1 v/v/v) were selected and evaluated for the extraction of insect lipids with DI-SPME fiber. Acetonitrile extraction offered the best qualitative, quantitative, and number of lipids extracted from insects samples results. Acetonitrile extracted high-boiling point compounds from both species of tested insects. The range of hydrocarbons was C25 (pentacosane) to C32 (dotriacontane) for T. castaneum and C26 (11-methylpentacosane) to C34 (tetratriacontane) for R. dominica. The major compounds extracted from the cuticular surface of T. castaneum were 11-methylheptacosane (20.71%) and 3-methylheptacosane (12.37%), and from R. dominica were 10-methyldotriacontane (14.0%), and 15-methyltritriacontane (9.93%). The limit of detection (LOD) for the n-alkane compounds ranged between 0.08 (nonacosane) and 0.26 (dotriacontane) µg/g and for the fatty acids between 0.65 (arachidic acid) to 0.89 (oleic acid) µg/g. The study indicated that DI-SPME GC-MS is a highly efficient extraction and a sensitive analytical method for the determination of non-derivatized insect lipids in cuticular and homogenized body tissues.