Identifying two moth species (Lepidoptera: Ditrysia) from Saudi Arabia using mitochondrial 16S rRNA sequences.

The mitochondrial genetic markers are considered useful tools for discrimination between more closely related lepidopteran taxa. Therefore, the present study aimed to investigate the role of mitochondrial (mt) 16 s rRNA gene in the determination of the taxonomic position for two moth species within Ditrysia clade. Maximum likelihood analysis has indicated a well-supported dendrogram based on the Tamura-Nei model for the recovered lepidopterans. The mt 16 s rRNA query sequences from 24 species within seven families were analyzed. This analysis and bootstrap confidence revealed two major clades representing Glossata suborder within Lepidoptera, with a close relationship of Noctuoidea + (Pyraloidea (Hesperioidea + Papilionoidea)). The subfamily Heliothinae forming a sister group with Risobinae (Noctinae + Hadeninae). In addition, there is a clear observation about the close relation between Phycitinae + Galleriinae within Pyraloidea and Cyrestinae + Limenitidinae within Papilionoidea. The present study supported that the Helicoverpa and Meroptera species are the first accounts of these genera inhabiting Saudi Arabia.

Farmers' knowledge, perception and management of mango mealy bug, Drosicha mangiferae Green (Hemiptera: Monophlebidae), on Mangifera indica in Punjab, Pakistan.

In 2004-05, during the December to February about 141 mango farmers were interviewed during peak activity of mango mealy bug in southern Punjab, Pakistan. The objective was to know the farmers' knowledge, perceptions and practices in the management of mango mealy bug. Most of the farmers (94.33%) reported that Chaunsa variety (king of all mango varieties) was susceptible to mango and irrigation water was the major source of flare up of this pest. Basudin and Supracide were the most commonly used insecticides as 72.92 and 51.77 percent farmers gave positive response and grease bands were applied for the control of mango mealy bug by the majority of the respondents. Hundred percent yield losses was told by 22.7 percent respondents whereas 75 percent, 50 percent and 25 percent losses were reported by 39.7, 31.9 and 14.2 insecticidal spray did not show satisfaction to the respondents for the control of fertilized females of mango mealy bug coming down from the trees. Lack of knowledge about the pest, lack of money, adulterated and shortage of pesticides, lack of unity amongst farmers Further the growers' views were tested in the field for confirmation and small land holdings were the main constraints for the control of mango mealy bug.

The phylogenetic position of Arhaphe deviatica within Hemipteran insects: A potential model species for eco-devo studies of symbiosis.

Insecta is known to be the most diverse group of species, exhibiting numerous forms of endosymbiotic associations. Molecular techniques have provided significant indicators for insect-microbe interactions. The present study aimed to register one of the true bugs of pentatomomorpha and clarify its taxonomic position through phylogenetic analysis of the partial 16S rRNA gene region. A maximum likelihood analysis retrieved a generally well-supported phylogeny based on Tamura 3-parameter model. Based on the partial mitochondrial 16S rRNA gene sequences, a phylogenetic study of suborder Heteroptera relationships within Hemipteras' order was constructed. Sequences of 221 bases of the 3' end of the gene from 28 species within 16 families were analyzed. This analysis and bootstrap confidence revealed two major clades comprising four suborders within Hemiptera, with a close relationship between Heteroptera + (Sternorrhyncha + (Auchenorrhycha + Coleorrhyncha)). Infraorder Pentatomomorpha is forming a sister group with a substantial bootstrap value to Cimicomorpha. Pyrrhocoroidea forms a sister relationship with Lygaeoidea + Coreoidea. There is a close relationship between Largidae and Pyrrhocoridae within Pyrrhocoroidea. The results show that the present species is firmly embedded in the genus Arhaphe with 94.35% sequence resemblance to its congeners. Besides, the recovered hemipteran species considered a potential model group for studying different symbionts. We propose both phylogenetic and ecological evolutionary developmental biology viewpoints for a more synthetic understanding of insect populations' molecular evolution.

Body Size and Cuticular Hydrocarbons as Larval Age Indicators in the Forensic Blow Fly, Chrysomya albiceps (Diptera: Calliphoridae).

Chrysomya albiceps (Wiedemann 1819) is one of the most important insects in forensic entomology. Its larval developmental and survival rates are influenced by nutritional resources, temperature, humidity, and geographical regions. The present study investigated the possibility of relying on body size and cuticular hydrocarbon composition as indicators for age estimation of the different larval instars of C. albiceps. Larvae were maintained in standardized laboratory conditions at different experimental temperatures. All larval instars (first, second, and third) were randomly collected for measuring their body sizes and for estimating their cuticular hydrocarbons at different rearing temperatures (30, 35, 40, and 45°C) using gas chromatography-mass spectrometry (GC-MS). Results indicated that the duration of larval stage was temperature dependent as it gradually decreased on increasing the rearing temperature (30, 35, and 40°C) except 45°C at which larval development was ceased. In contrary, larval body size, in terms of length, width, and weight, was temperature dependent as it gradually increased with larval development on increasing rearing temperature except at 45°C at which larval development was ceased. The GC-MS showed a significant difference in the extracted components of cuticular hydrocarbons between different larval instars reared in the same temperature and between the same larval instar that reared at different temperatures. Furthermore, the highest and lowest amounts of cuticular hydrocarbons were detected at 35 and 40°C, respectively. Overall, larval body size and cuticular hydrocarbon components were temperature dependent within the range 30-40°C, which may suggest them as possible reliable age indicators for estimating the postmortem interval in the field of medicolegal entomology.

Gene expression as age estimation marker in the larval stages of the forensic blowfly, Chrysomya albiceps, at different temperatures.

The blowfly Chrysomya albiceps (Diptera: Calliphoridae) has been known to breed in both animal and human carcasses in different geographical regions and is considered an important post-mortem indicator in forensic entomology. Determining the age of its larvae is an essential tool for the accurate determination of post-mortem intervals (PMI). This study adopted a molecular-based approach for age determination of the immature stages of the blowfly, C. albiceps, by evaluating the genes 15_2, 2014192, EcdR, AR, hsp90, and Actin, during larval development at different temperatures. Targeted genes were amplified by real-time PCR (RT-PCR), and the resulting amplicons were normalized against the two housekeeping genes, rp49 and 19_2. Data showed that the expression profile of AR was constant throughout all larval developmental stages at all tested temperatures. 2014192 showed low expression levels throughout the 1st, 2nd, and 3rd larval stages. Hsp90 and EcdR showed different expression profiles. The expression of 15_2 was low at the tested temperatures and was undetectable on most days. Collectively, the results of this study indicate that larvae exhibit temperature-dependent and age-specific up- and down-regulation in the expression profile of the targeted genes. This may indicate a possibility to be utilized as age estimation markers for C. albiceps.

Seasonal Impact of Heroin on Rabbit Carcass Decomposition and Insect Succession.

This study investigates the succession of insects and the length of the decomposition of rabbit carcasses injected with heroin. In total, 24 rabbits (2-2.5 kg, Oryctolagus cuniculus domesticus L.) were used in the study. Twelve carcasses were exposed to insect activity in winter (January and February) and 12 in summer (July and August) in Riyadh, Saudi Arabia. Three replicate experiments were conducted in each season. For each experiment, three rabbits were injected with varying doses of heroin, and the fourth rabbit was used as a control, being injected only with saline solution. Insects belonging to the order Diptera, Coleoptera, and Hymenoptera were attracted to the carcasses. In both of the investigated seasons, heroin appeared to delay the decomposition process but did not have a significant impact on the number of insects. In general, the number of flies in both seasons was substantially higher than that of beetles and ants. During the summer, flies were more attracted to treated carcasses with a higher dose of heroin. Some flies such as Musca domestica L. (Diptera: Muscidae) and Chrysomya albiceps Wiedemann (Diptera: Calliphoridae) and some beetles such as Pimelia boyeri Solier (Coleoptera: Tenebrionidae) were represented on the carcasses by different developmental instars (larvae, pupae, and adults). Overall, heroin has not potential effects on the insect succession patterns of insect, however, influences the decomposition of rabbit carcasses. These findings are important while using insects in medicolegal investigation, since heroin is one of the common poisons used as an addictive agent.

Molecular phylogeny of Monomorium pharaonis (Hymenoptera: Formicidae) based on rRNA sequences of mitochondrial gene.

Many studies on forensic entomology have focussed on Diptera and lack the Hymenopteras population. Nonetheless, hymenopterans are part of the entomofaunal colonization of a corpse. Morphologically, it is difficult to identify and distinguish between them. In this study, using mitochondrial DNA knowledge, the molecular analysis was performed to classify the recovered species of hymenoptera collected from rabbit carcass, quickly and accurately. A molecular identification method with a 251-bp fragment of the 16S ribosomal gene RNA (16S rRNA) from a single ant species was evaluated. The maximum likelihood method analysis has recovered a generally well-supported phylogeny, with most taxa and species groups currently being recognized as monophyletic. The aculeate consistsof some Hymenoptera's best known. Their sister group has traditionally been considered in Ichneumonoidea. In addition, Trigonaloidea was found as the aculeates' sister group and Crabronidae in Apoidea forming the Formicidae's sister group. These results will play an important role in the implementation of the Saudi database forensically relevant ants.

A Comparative Study of Insect Succession on Rabbit Carrion in Three Different Microhabitats.

Insect succession on three rabbit carcasses placed 110.49 cm above ground in winter in Riyadh City, Saudi Arabia was examined in three different microhabitats: dry, semi-submerged, and submerged in tap water. The complete decomposition of the carcasses took 20 d and included five stages. In total 715 insects collected from carrion in different microhabitats were identified morphological, followed by the partial sequence confirmation of their 16S rRNA mitochondrial genes. Results showed that the insect species found and species richness differed among microhabitats. Carrion placed in the dry microhabitat showed the highest species richness and colonized by 400 insect specimens belonging to 16 species. Carrion placed in the semi-submerged microhabitat contained 271 insect specimens belonging to 12 species, showing a relatively low species richness. Lastly, the submerged microhabitat showed the lowest species richness, as the carrion placed in it attracted only 44 insects belonging to 9 species.