Galeruca daurica (Joannis) is a new outbreak pest in the Inner Mongolia grasslands in northern China. Heat shock protein 10 and 60 (Hsp10 and Hsp60) genes of G. daurica, designated as GdHsp10 and GdHsp60, were cloned by rapid amplification of cDNA ends techniques. Sequence analysis showed that GdHsp10 and GdHsp60 encoded polypeptides of 104 and 573 amino acids, respectively. Sequence alignment and phylogenetic analysis clearly revealed that the amino acids of GdHsp10 and GdHsp60 had high homology and were clustered with other Hsp10 and Hsp60 genes in insects which are highly relative with G. daurica based on morphologic taxonomy. The mRNA expression analysis by real-time PCR revealed that GdHsp10 and GdHsp60 were expressed at all development stages and in all tissues examined, but expressed highest in eggs and in adults’ abdomen; both heat and cold stresses could induce mRNA expression of GdHsp10 and GdHsp60 in the 2nd instar larvae; the two Hsp genes were expressed from high to low with the extension of treatment time in G. daurica eggs exposed to freezing point. Overall, our study provides useful information to understand temperature stress responses of Hsp60 and Hsp10 in G. daurica, and provides a basis to further study functions of Hsp60/Hsp10 relative to thermotolerance and cold hardiness mechanism.

Natural enemies are exposed to insecticide sprays for herbivorous species and may evolve field resistance to insecticides. Natural enemies selected for resistance in the field, however, are welcome for pest control. The susceptibility of 20 populations of Eriopis connexa from various crop ecosystems to λ-cyhalothrin was tested. Three bioassays were conducted: (i) topical treatment with lethal dose (LD)50 previously determined for populations considered standard for susceptibility (LD50S) and for resistance (LD50R) to λ-cyhalothrin at technical grade; (ii) dose–mortality assay to calculate the LD for populations exhibiting significant survival to the LD50R; and (iii) determination of survival when exposed to dried residues at field rates. Among the 20 tested populations, seven populations did not survive or survival rates were lower than 10% when treated with LD50R; three populations survived >20%, but lower than 50%; while ten populations exhibited equal or greater survival rates compared with the 50% expected survival for the LD50R. Thus, these ten populations were subjected to dose–mortality response, and the LD50 values varied from 0.046 to 5.44 µg a.i./insect with resistance ratio of 8.52- to 884.08-folds. Adults from these ten populations that were ranked as resistant according to the LD50R exhibited survival from 44.5 to 100% exposed to the lowest and from 38.8 to 100% exposed to the highest field rates of λ-cyhalothrin, respectively. Otherwise, the remaining ten populations ranked as susceptible according to the LD50R showed survival from 3.3 to 56% exposed to the lowest and from 0 to 17.7% exposed to the highest field rates of λ-cyhalothrin, respectively. Therefore, 50% of the tested E. connexa populations exhibited field-evolved resistance to λ-cyhalothrin and the use of a discriminatory LD50 for resistance matched the survival obtained when exposed to the insecticide field rates.

The preference-performance hypothesis (PPH) has widely been used to explain host exploitation patterns by phytophagous insects. However, this hypothesis often fails in the case of polyphagous species when compared with specialists. One explanation, validated by the information-processing hypothesis (IPH), considers that polyphagous insects are unable to process a large array of cues, which hinders females from distinguishing between high- and low- quality hosts. Here we analyzed Anastrepha ludens female host preference and offspring performance, and tested if neuronal limitations could possibly play a role in the incapacity of the polyphagous A. ludens to make ‘accurate decisions’ and therefore partially explain mismatches related to PPH. Results testing the PPH by correlating female preference to six naturally occurring hosts and its offspring outcomes show that A. ludens females oviposited greater proportions of eggs on fruit according to hierarchical preferences. Infestation level was low in white sapote, the preferential and seemingly putative ancestral host, likely due to sapote defence mechanisms. Pupal weight and adult size were lower when A. ludens larvae developed in guava (conditional host that was artificially infested) and peach, a lower ranked host compared with ‘Marsh’ grapefruit, white sapote, and ‘Manila’ mango (three preferred hosts). Larvae reared in ‘Manzano’ pepper, a low-ranked host, performed better than in peach and guava. Results testing the IPH, show that polyphagous A. ludens females were less accurate when discerning between a non natural host (guava) when compared with a preferred, natural host (grapefruit): error rate was significantly higher, number of oviposited fruit in a 6-h period was extremely low, time searching and ovipositing took longer, and pupae recovery was extremely low. Our findings indicate that both hypotheses tested are complementary and help better understand host use by A. ludens. However, we also discuss the complexity of polyphagy considering other factors such as plant resistance/defence mechanisms which are not fully addressed in both theories tested.

Thiamine is a vitamin that has been shown to act as a trigger to activate plant defence and reduce pathogen and nematode infection as well as aphid settling and reproduction. We have here investigated whether thiamine treatments of seeds (i.e. seed dressing) would increase plant resistance against aphids and whether this would have different effects on a generalist than on specialist aphids. Seeds of wheat, barley, oat and pea were treated with thiamine alone or in combination with the biocontrol bacteria Pseudomonas chlororaphis MA 342 (MA 342). Plants were grown in climate chambers. The effects of seed treatment on fecundity, host acceptance and life span were studied on specialist aphids bird cherry-oat aphid (Rhopalosiphum padi L.) and pea aphid (Acyrthosiphon pisum Harris) and on the generalist green peach aphid (Myzus persicae, Sulzer). Thiamine seed treatments reduced reproduction and host acceptance of all three aphid species. The number of days to reproduction, the length of the reproductive life, the fecundity and the intrinsic rate of increase were found reduced for bird cherry-oat aphid after thiamine treatment of the cereal seeds. MA 342 did not have any effect in any of the plant-aphid combinations, except a weak decrease of pea aphid reproduction on pea. The results show that there are no differential effects of either thiamine or MA 342 seed treatments on specialist and generalist aphids and suggest that seed treatments with thiamine has a potential in aphid pest management.

Rapid determination of the larval species composition and understanding of their genetic structure is important to establish the appropriate management system for multiple species infesting in fruits. We established accurate and rapid diagnostic methods based on multiplex polymerase chain reaction (PCR) diagnostic techniques to discriminate the three major lepidopteran species in orchard, Carposina sasakii, Grapholita dimorpha, and Grapholita molesta. Each species was identified by amplifying species-specific PCR products (375 bp for C. sasakii, 125 and 234 bp for G. dimorpha, and 125 bp for G. molesta). Based on species composition analysis from six types of infested fruits, G. dimorpha constituted the highest proportion (47.8%), followed by 35.2 and 13.5% for G. molesta and C. sasakii, respectively. Interestingly, high prevalence was found in G. dimorpha and G. molesta for plum and peach, respectively. Based on genetic diversity analysis, the three insect species exhibited moderate or high haplotype diversity and low nucleotide diversity, ranging from 0.319 to 0.699 and 0.0006 to 0.0045, respectively. Demographic expansion was not detected according to either a neutrality test or mismatch distribution analysis. Moreover, no significant genetic structure corresponding to province, host plant, fruit type, or collection period was observed. These results suggest that the population of each species would have high dispersal ability following fruit-generating periods via intrinsic host adaptation ability regardless of the spatial and temporal conditions. Determination of larval composition on fruit is valuable for establishing appropriate management systems that take the species into consideration; additionally, population genetic approaches can be utilized to understand the effects of environmental factors (province, host fruit, fruit type, etc.) on population structures.

A recent study on ladybird, Menochilus sexmaculatus (Fabricius) demonstrates that males perform post-copulatory mate guarding in the form of prolonged mating durations. We investigated whether food resource fluctuation affects pre- and post-copulatory behaviour of M. sexmaculatus. It has not been studied before in ladybirds. For this, adults were subjected to prey resource fluctuations sequentially at three levels: post-emergence (Poe; 10 days), pre-mating (Prm; 24 h) and post-mating (Pom; 5 days; only female). The food resource conditions at each level could be any one of scarce, optimal or abundant. Pre-copulatory and post-copulatory behaviour, and reproductive output were assessed. Post-emergence and pre-mating nutrient conditions significantly influenced the pre-copulatory behaviour. Males reared on scarce post-emergence conditions were found to require significantly higher number of mating attempts to establish mating unlike males in the other two food conditions. Under scarce post-emergence and pre-mating conditions, time to commencement of mating and latent period were high but opposite result was obtained for mate-guarding duration. Fecundity and per cent egg viability were more influenced by post-mating conditions, with scarce conditions stopping oviposition regardless of pre-mating and post-emergence conditions. Present results indicate that pre- and post-copulatory behaviour of ladybird is plastic in nature in response to food resource fluctuations.

RNA interference (RNAi) is a very effective technique for studying gene function and may be an efficient method for controlling pests. Trehalose-6-phosphate synthase (TPS), which plays a key role in the synthesis of trehalose and insect development, was cloned in Tribolium castaneum (Herbst) (TcTPS) and the putative functions were studied using RNAi via the injection of double-stranded RNA (dsRNA) corresponding to conserved TPS and trehalose-6-phosphate phosphatase domains. Expression analyses show that TcTPS is expressed higher in the fat body, while quantitative real-time polymerase chain reaction results show that the expression of four trehalase isoforms was significantly suppressed by dsTPS injection. Additionally, the expression of six chitin synthesis-related genes, such as hexokinase 2 and glutamine-fructose-6-phosphate aminotransferase, was suppressed at 48 and 72 h post-dsTPS-1 and dsTPS-2 RNA injection, which were two dsTPS fragments that had been designed for two different locations in TcTPS open reading frame, and that trehalose content and trehalase 1 activity decreased significantly at 72 h post-dsRNA injection. Furthermore, T. castaneum injected with dsTPS-1 and dsTPS-2 RNA displayed significantly lower levels of chitin and could not complete the molting process from larvae to pupae, revealing abnormal molting phenotypes. These results demonstrate that silencing TPS gene leads to molting deformities and high mortality rates via regulation of gene expression in the chitin biosynthetic pathway, and may be a promising approach for pest control in the future.

Characterization and gene cloning of acetylecholinesterase (AChE) in the insecticide-resistant (R) and -susceptible (S) insects have been reported in the past. However, the studies focused mostly on herbivorous pests, rather than predacious species, such as ladybird beetles. Using R and S Propylaea japonica (thunberg), a full-length cDNA sequence (2928 bp) of the ace1-type AChE gene was determined for the first time. The ace1 encoding a protein of 645 amino acids contained typical conserved motifs, such as FGESAG domains, catalytic triad, acyl pocket, oxyanino hole, choline binding site, peripheral anionic site, omega loop and conserved aromatic residues. R P. japonica displayed 50-times greater resistance to chlorpyrifos or mathamidophos with a significantly lower AChE sensitivity to paraoxon, malaoxon, chlorpyrifos or methamidophos than its S counterpart. Five amino acids in the ace1 of R P. japonica differed from those found in S P. japonica. One of them, F358S, located in the acyl-binding pocket, might play a crucial role in the resistance of the insect to organophosphates (OPs). Whereas, K493E and I538V, which were close to some of the conserved aromatic amino acids (i.e., H509, Y511, and W499) in the gorge, and G571R and T576A near C593 that formed the disulfide bonds with C471, might also involve in the change of insecticide resistance in P. japonica. AChE insensitivity and amino acid replacements, particularly F358S, might be the determining factors in the alteration of OPs-resistance in P. japonica.

The Red Palm Weevil (RPW) Rhynchophorus ferrugineus (Olivier 1790) is an invasive pest from southeastern Asia and Melanesia that in the last 30 years has spread widely in the Middle East and Mediterranean Basin. Its stem-boring larvae cause great damage to several palm species of the Arecaceae family, many of which are economically important for agricultural and ornamental purposes. Therefore, great attention has recently been focused in studying this species to identify sustainable and effective eradication strategies, such as sterile insect technique (SIT). The rapid spread of RPW is associated with its high reproductive success. To evaluate the suitability of a SIT strategy, particular physiological and behavioral aspects of RPW reproduction, such as the presence of polyandry and post-copulatory sperm selection mechanisms, were investigated. To determine paternity of progeny from multiply mated females, double-crossing experiments were carried out confining individual females with either a wild-type male or a γ-irradiated male (Co-60). Fecundity and fertility of females were scored to evaluate post-copulatory sperm selection. Results showed that progeny were almost exclusively produced by the sperm of the second male, suggesting that a last-male sperm precedence is expressed at high levels in this species, and providing interesting insights for an area-wide RPW management strategy such as the SIT.

We examined the life history consequences of cornicle secretion by Aphis fabae Scopoli in second and fourth instars, and its effects on host suitability for its parasitoid, Lysiphlebus fabarum (Marshall). Cornicle secretion did not affect aphid fecundity, but secretion in the second instar enhanced life table parameters, whereas secretion in the fourth instar affected them negatively, suggesting a higher cost of secretion in later instars. Secretion in either instar improved host suitability for L. fabarum. Although control and treated aphids were parasitized at similar rates, and with similar success, wasps developed faster and emerged as larger adults in aphids that had secreted, regardless of instar. Transgenerational effects were also evident. Progeny emergence was higher when parental wasps developed in fourth instars than in seconds, whether aphids secreted or not, and progeny were larger when parental hosts secreted in the second instar, but not in the fourth. Secreting fourth instars were preferred to controls by L. fabarum females in choice tests, but not secreting second instars, and fourth-instar secretion improved wasp emergence. When control aphids were attacked, second instars were more likely to secrete than fourth instars, whereas the latter were more likely to kick the parasitoid. Cornicle secretion reduced the probability of subsequent secretion events and the frequency of other aphid defensive behaviors, indicating energetic tradeoffs among defensive tactics. Overall, our results revealed that cornicle secretion by immature A. fabae exacts both physiological and behavioral costs and results in improved host suitability for its parasitoid.

It is difficult to control Holotrichia parallela Motschulsky with chemical insecticides due to the larvae's soil-living habit, thus the pest has caused great economic losses in agriculture. In addition, uridine diphosphate-glycosyltransferases (UGTs) catalyze the glycosylation process of a variety of small lipophilic molecules with sugars to produce water-soluble glycosides, and play multiple roles in detoxification, endobiotic modulation, and sequestration in an insect. Some UGTs were found specifically expressed in antennae of Drosophila melanogaster and Spodoptera littoralis, and glucurono-conjugated odorants could not elicit any olfactory signals, suggesting that the UGTs may play roles in odorant inactivation by biotransformation. In the current study, we performed a genome-wide analysis of the candidate UGT family in the dark black chafer, H. parallela. Based on a UGT gene signature and the similarity of these genes to UGT homologs from other organisms, 20 putative H. parallela UGT genes were identified. Bioinformatics analysis was used to predict sequence and structural features of H. parallela UGT proteins, and revealed important domains and residues involved in sugar donor binding and catalysis by comparison with human UGT2B7. Phylogenetic analysis of these 20 UGT protein sequences revealed eight major groups, including both order-specific and conserved groups, which are common to more than one order. Of these 20 UGT genes, HparUGT1265-1, HparUGT3119, and HparUGT8312 were highly (>100-fold change) expressed in antennae, suggesting a possible role in olfactory tissue, and most likely in odorant inactivation and olfactory processing. The remaining UGT genes were expressed in all tissues (head, thorax, abdomen, leg, and wing), indicating that these UGTs likely have different biological functions. This study provides the fundamental basis for determining the function of UGTs in a highly specialized olfactory organ, the H. parallela antenna.

European dry-wood termites belong to the genus Kalotermes (Kalotermitidae), one of the two termite genera in Europe. Until the recent description of two new species, Kalotermes italicus in Italy and Kalotermes phoenicae in the eastern Mediterranean area, Kalotermes flavicollis was the only taxon known in this region. The presence of additional entities, suggested by morphological and physiological variation observed in K. flavicollis, was supported by molecular studies revealing four distinct genetic lineages: lineage A, K. flavicollis sensu strictu, from the Aegean area to Italy; lineage B, in Tuscany; lineage SC, in Sardinia and Corsica; lineage SF, in southern France. Lineages A and B may form mixed colonies, suggesting hybridization. To draw a more detailed picture of Kalotermes evolution and biogeography in Europe, we analyzed samples from previously unsampled areas, such as Spain and southern Italy, by means of the highly informative cox1/trnL/cox2 mitochondrial DNA marker. Overall, phylogenetic analyses confirmed previously identified lineages and taxa, but widened the distribution of the lineage SC to the mainland and of the lineage SF to Spain and Portugal. Results further provided evidence for the synonymy between lineage B and K. italicus. Species delimitation analysis suggested that the three K. flavicollis lineages, as well as K. italicus, can be separate taxa. Data also suggest a possible interspecific hybridization between K. italicus and both K. flavicollis lineages A and SC.

Aphelinus mali (Haldeman) is an effective natural enemy of woolly apple aphid (WAA), Eriosoma lanigerum (Hausmann). Previous studies have found that, with WAA from Shandong Province (Qingdao) as the host, there are significant differences in various biological characteristics between a Shandong clade and Liaoning clade of A. mali. The ability of the Shandong clade to control this aphid was significantly higher than that of the Liaoning clade in Shandong Province. In order to determine whether differences were caused by better adaptation of the Shandong parasitoid clade to the population of the host in that province or if it represents a more general fitness of this clade to control the host regardless of location, we compared the same parasitoid clades with hosts from Hebei Province. We found no significant differences in the developmental threshold temperature, effective accumulated temperature, fecundity, longevity, and oviposition period of the two clades, but the duration of host searching of the Shandong clade was significantly longer than that of the Liaoning clade. The instantaneous attack rate, the control ability (a/Th), the search parameter (Q) of the Shandong clade (0.0946, 0.543, 0.0725) of A. mali were higher than that of the Liaoning clade (0.0713, 0.382, 0.0381), and therefore, with WAA from Hebei Province as the host, the host adaptability of the Shandong clade of A. mali was not worse than that of the Liaoning clade, while the pest control ability of the Shandong clade was still greater than that of the Liaoning clade.

The effect of increasing mating delay on the reproductive performance and population growth rates of the vine mealybug, Planococcus ficus (Signoret) (Hemiptera: Pseudococcidae), was investigated under laboratory conditions. Virgin females were mated at 1, 3, 5, 7, 14, 21 and 28 days after emergence and reproductive and life table parameters were estimated. The pre-oviposition period (number of days between mating and the onset of oviposition) significantly decreased in females mated within 7 days, whereas females mated at older ages showed equivalent pre-oviposition periods (<4 days). The length of the oviposition period did not vary with increasing age at mating. Female longevity significantly increased in females mated at 21 and 28 days, as a consequence of a longer pre-reproductive period. Fecundity and sex ratio were not affected by the female age at mating, whereas fertility was higher in mealybugs mated at older ages. Additional field observations highlighted that young and old virgin females were equally able to attract males, as both mated on the same day as the field release. Mating delay also affected the life table parameters of P. ficus, as the intrinsic and finite rates of increase did not differ in mealybugs mated within 7 days and significantly decreased in females mated at older ages. The mean generation time and the population doubling time were overall similar in females mated at 1–7 days, and increased significantly in females that experienced longer mating delays. In terms of the mating disruption control of P. ficus, our findings indicate that this method would be effective if mating is delayed >7 days, as shorter delays in mating did not reduce the population growth rates.

Over the last 40 years, many types of population genetic markers have been used to assess the population structure of the pest moth species Helicoverpa armigera. While this species is highly vagile, there is evidence of inter-continental population structure. Here, we examine Z-chromosome molecular markers within and between Chinese and Australian populations. Using 1352 polymorphic sites from 40 Z-linked loci, we compared two Chinese populations of moths separated by 700 km and found virtually no population structure (n = 41 and n = 54, with <1% of variation discriminating between populations). The levels of nucleotide diversity within these populations were consistent with previous estimates from introns in Z-linked genes of Australian samples (π = 0.028 vs. 0.03). Furthermore, all loci surveyed in these Chinese populations showed a skew toward rare variants, with ten loci having a significant Tajima's D statistic, suggesting that this species could have undergone a population expansion. Eight of the 40 loci had been examined in a previous study of Australian moths, of which six revealed very little inter-continental population structure. However, the two markers associated with the Cyp303a1 locus that has previously been proposed to be a target of a selective sweep, exhibited allele structuring between countries. Using a separate dataset of 19 Australian and four Chinese moths, we scanned the molecular variation distributed across the entire Z-chromosome and found distinct blocks of differentiation that include the region containing Cyp303a1. We recommend some of these loci join those associated with insecticide resistance to form a set of genes best suited to analyzing population structure in this global pest.

The aim of this study was to verify the occurrence of the coffee berry borer, Hypothenemus hampei (Ferrari), and to evaluate the population fluctuation of the pest in the Brazilian Cerrado (Federal District). The study was conducted, between November 2014 and October 2015, at Embrapa Cerrados (Planaltina/DF, Brazil) in an irrigated conilon coffee production area. In November 2014, 120 samples (ten berries/sample) were collected from berries that had fallen on the ground from the previous harvest. Between November 2014 and October 2015, insects were collected weekly, using traps (polyethylene terephthalate bottles) baited with ethyl alcohol (98 GL), ethyl alcohol (98 GL) with coffee powder, or molasses. Between January and July 2015, samples were collected fortnightly from 92 plants (12 berries per plant). All samples were evaluated for the presence of adult coffee berry borers. Samples from the previous harvest had an attack incidence of 72.4%. The baited traps captured 4062 H. hampei adults, and showed no statistical difference in capture efficiency among the baits. Pest population peaked in the dry season, with the largest percentage of captured adults occurring in July (31.0%). An average of 18.6% of the collected berries was attacked by the borer and the highest percentage incidence was recorded in July (33.2%). Our results suggest that the coffee berry borer, if not properly managed, could constitute a limiting factor for conilon coffee production in the Brazilian Cerrado.

Natural control by predators and parasitoids provides an important and often unnoticed ecosystem service to agricultural landscapes by reducing pest populations in crops. The current model of horticultural intensification in south-eastern Spain produces high yields but has also resulted in a landscape almost completely covered by plastic. Promoting natural areas among greenhouses could enhance biodiversity, by being beneficial insects, and reduce pest pressure outdoors. The first step is to ascertain how pests and their natural enemies (NEs) use Mediterranean vegetation for selecting the best plants for pest suppression outdoors. The abundance of the two major horticultural pests, the tobacco whitefly, Bemisia tabaci, and the western flower thrips, Frankliniella occidentalis, together with their NEs, were assayed in 22 flowering perennial plants, which were newly planted in an experimental field surrounded by greenhouses. Eight plant species were identified as the most critical species for sustaining pest populations outdoors. A set of five plant species supported a medium level of pests, and another set of ten plant species supported the lowest level of both pests. Tobacco whitefly occurred in a few plants species, whereas western flower thrips occurred on almost all the plant species studied, and was favoured by the presence of flowers in perennial plants. The results suggest that plant diversity may provide relatively few acceptable host plants for tobacco whitefly than for western flower thrips. NEs were generally collected in plants that also supported abundance of pests, indicating that host/prey availability, more than food resources from flowers, was a stronger predictor of NE abundance in perennial plants. Field trials using the plants with the lowest host acceptance by pests are needed in order to ascertain whether pest abundance outdoors is reduced.

The subterranean termite Reticulitermes grassei Clément causes lesions in the trunk of Quercus suber L. by constructing feeding galleries, but no information is available regarding other Quercus species from the Mediterranean region. This work aimed to study the suitability of the other main oak species of Mediterranean forests as a food resource for R. grassei. Two experiments, choice and non-choice feeding, were conducted lasting for 15, 30, and 45 days each. In the non-choice experiment, termites were offered one of the following food types: Quercus suber, Quercus ilex L., Quercus faginea Lam, cork or Pinus pinea L., which was considered the control. The choice feeding experiment used all the same food types listed above, supplied simultaneously in the same container. Food selection was examined by analysing the relationships over time between surviving termites and food consumption. The results indicated that R. grassei could be considered a generalist species, as it consumed the cork and wood of all oak species, as well as displaying a clear preference for soft wood (pine). Correlation analysis indicated that consumption was not dependent on wood density. Survival of R. grassei was influenced by the time of exposure to different oak species, but a high survival rate was maintained over time in the pine treatment (upper 70% in the three experiments). Given these results, it can be concluded that all the oak species are a suitable food source for R. grassei.

A wide variety of abiotic and biotic factors act on insect pests to regulate their populations. Knowledge of the time and magnitude of these factors is fundamental to understanding population dynamics and developing efficient pest management systems. We investigate the natural mortality factors, critical pest stages, and key mortality factors that regulate Chrysodeixis includens populations via ecological life tables. The total mortality caused by natural factors was 99.99%. Natural enemies were the most important mortality factors in all pest stages. The critical stages of C. includens mortality were second and fourth instars. The key mortality factors were predation by ants in the second instar and predation by Vespidae in the fourth instar. The elimination of these factors can cause an increase of 77.52 and 85.17% of C. includens population, respectively. This study elucidates the importance of natural enemies and other natural mortality factors in C. includens population regulation. These factors should be considered in developing and implementing C. includens management strategies and tactics in order to achieve effective and sustainable pest control.

The date palm hopper, Ommatissus lybicus, is a key pest of the date palm, which is expected to be comprised of many allopatric populations. The current study was carried out to determine bacterial endosymbiont diversity in the different populations of this pest. Ten date palm hopper populations were collected from the main date palm growing regions in Iran and an additional four samples from Pakistan, Oman, Egypt and Tunisia for detection of primary and secondary endosymbionts using polymerase chain reaction (PCR) assay with their specific primers. The PCR products were directly sequenced and edited using SeqMan software. The consensus sequences were subjected to a BLAST similarity search. The results revealed the presence of ‘Candidatus Sulcia muelleri’ (primary endosymbiont) and Wolbachia, Arsenophonus and Enterobacter (secondary endosymbionts) in all populations. This assay failed to detect ‘Candidatus Nasuia deltocephalinicola’ and Serratia in these populations. ‘Ca. S. muelleri’ exhibited a 100% infection frequency in populations and Wolbachia, Arsenophonus and Enterobacter demonstrated 100, 93.04 and 97.39% infection frequencies, respectively. The infection rate of Arsenophonus and Enterobacter ranged from 75 to 100% and 62.5 to 100%, respectively, in different populations of the insect. The results demonstrated multiple infections by ‘Ca. Sulcia muelleri’, Wolbachia, Arsenophonus and Enterobacter in the populations and may suggest significant roles for these endosymbionts on date palm hopper population fitness. This study provides an insight to endosymbiont variation in the date palm hopper populations; however, further investigation is needed to examine how these endosymbionts may affect host fitness.