BackgroundThe predatory mirids of the genus Macrolophus are key natural enemies of various economically important agricultural pests. Both M. caliginosus and M. pygmaeus are commercially available for the augmentative biological control of arthropod pests in European greenhouses. The latter species is known to be infected with Wolbachia -inducing cytoplasmic incompatibility in its host- but the presence of other endosymbionts has not been demonstrated. In the present study, the microbial diversity was examined in various populations of M. caliginosus and M. pygmaeus by 16S rRNA sequencing and denaturing gradient gel electrophoresis.ResultsBesides Wolbachia, a co-infection of 2 Rickettsia species was detected in all M. pygmaeus populations. Based on a concatenated alignment of the 16S rRNA gene, the gltA gene and the coxA gene, the first is phylogenetically related to Rickettsia bellii, whereas the other is closely related to Rickettsia limoniae. All M. caliginosus populations were infected with the same Wolbachia and limoniae-like Rickettsia strain as M. pygmaeus, but did not harbour the bellii-like Rickettsia strain. Interestingly, individuals with a single infection were not found. A PCR assay on the ovaries of M. pygmaeus and M. caliginosus indicated that all endosymbionts are vertically transmitted. The presence of Wolbachia and Rickettsia in oocytes was confirmed by a fluorescence in situ hybridisation. A bio-assay comparing an infected and an uninfected M. pygmaeus population suggested that the endosymbionts had minor effects on nymphal development of their insect host and did not influence its fecundity.ConclusionTwo species of the palaearctic mirid genus Macrolophus are infected with multiple endosymbionts, including Wolbachia and Rickettsia. Independent of the origin, all tested populations of both M. pygmaeus and M. caliginosus were infected with three and two endosymbionts, respectively. There was no indication that infection with endosymbiotic bacteria had a fitness cost in terms of development and fecundity of the predators.

Egg parasitoids of the genus (Hymenoptera: Trichogrammatidae) are important natural enemies of many lepidopterous pests in agricultural and forest areas. Because the use of indigenous populations/species can significantly increase the biological control efficiency, the characterization of endemic populations and precise species identification is important. In this study, wasps were collected from parasitized eggs of several lepidopteran pests in Northern and Central Iran. Seven species were identified based on the morphology and the nucleotide sequence, size and restriction profile (PCR-RFLP) of the internal transcribed spacer 2 (ITS2) region of the rDNA of . Bezdenko, Marchal, Hartig, Westwood, Girault, Voegel, and Sorokina. This is the first report of in Iran. Additionally, natural populations were surveyed for the prevalence of . Two populations of were found to be infected by a single strain which was identified by using a multi-locus sequencing approach

Macrolophus pygmaeus is a heteropteran predator that is widely used in European glasshouses for the biological control of whiteflies, aphids, thrips and spider mites. We have demonstrated that the insect is infected with the endosymbiotic bacterium Wolbachia pipientis. Several gene fragments of the endosymbiont were sequenced and subsequently used for phylogenetic analysis, revealing that it belongs to the Wolbachia supergroup B. The endosymbiont was visualized within the ovarioles using immunolocalization. Tetracycline treatments were used to cure M. pygmaeus from its infection. Although a completely cured line could not be obtained by this approach, the application of a constant antibiotic pressure over 13 generations resulted in a line with a significantly reduced Wolbachia concentration. Crosses performed with this tetracycline-treated line revealed that the endosymbiont causes severe cytoplasmic incompatibility. This is the first report of a reproductive effect induced by Wolbachia in an economically important heteropteran predator that may have vital implications for its commercial production and use in biological control.

The generalist predator Macrolophus pygmaeus Rambur (Hemiptera: Miridae) is a key biological control agent in European greenhouses. The influence of acclimation, infection with endosymbiotic bacteria and diet on the cold tolerance of the Mediterranean biocontrol population was assessed by determining the supercooling point, i.e. the temperature at which the insect's body fluids freeze. This parameter provides a first indication of an insect's establishment potential in a new region and of its possible geographical range. Allowing the predatory bugs to adapt to lower temperatures resulted in an increase in supercooling ability. Macrolophus pygmaeus bugs exposed to antibiotics in their artificial diet and hence cured from their infection with the endosymbiotic bacteria Wolbachia pipientis, Rickettsia bellii and R. limoniae were more tolerant to freezing than infected bugs. The diet of the predators also affected the freezing temperature of the body fluids. Predators fed an artificial diet based on egg yolk were less resistant to freezing than those fed Ephestia kuehniella Zeller (Lepidoptera: Pyralidae) eggs. These findings illustrate that several factors may affect the cold hardiness of a biocontrol agent and may thus complicate the evaluation of its establishment potential in the framework of an environmental risk assessment