Cockroach Blattella Germanica Research Articles

Synergistic effect of piperonyl butoxide on the toxicity of alpha-cypermethrin and deltamethrin against pyrethroid-resistant german cockroach Blattella germanica (Blattodea: Ectobiidae) strains in Turkey

Synergistic effect of piperonyl butoxide on the toxicity of alpha-cypermethrin and deltamethrin against pyrethroid-resistant german cockroach Blattella germanica (Blattodea: Ectobiidae) strains in Turkey

Broad complex and wing development in cockroaches

In hemimetabolan insects, the transcription factor Broad complex (Br–C) promotes wing growth and development during the nymphal period. We wondered whether Br–C could trigger the initiation of wing development, using the cockroach Blattella germanica as a model. We show that first instar nymphs have their unique identity of these three thoracic segments specified. During embryogenesis, the expression of Br–C and some wing-related genes show two matching waves. The first takes place before the formation of the germ band, which might be involved in the establishment of various developmental fields including a potential “wing field”, and the second wave around organogenesis, possibly involved in the initiation of wing development. However, the expression of Br–C in early embryogenesis concentrates in the developing central nervous system, thus not co-localizing with the expression of the typical wing-related gene vestigial, which is expressed at the edge of the thoracic and abdominal segments. This suggests that Br–C is not specifically involved in the establishment of a potential “wing field” in early embryogenesis. Moreover, maternal RNAi for Br–C depletes the first wave of Br–C expression but does not affect the early expression of wing-related genes. As maternal Br–C RNAi did not deplete the second expression wave of Br–C, we could not evaluate if Br–C is involved in the initiation of wing development. Alternatively, using nymphal RNAi of Br–C and Sex combs reduced (Scr), we show that Br–C contributes to the formation of ectopic wing structures that develop in the prothorax when Scr is depleted. The gene most clearly influenced by Br–C RNAi is nubbin (nub), which, in nymphs is crucial for wing growth. Together, these results suggest that Br–C does not specifically contribute to the establishment of the “wing field”, but it does seem important later, in the initiation of wing development, enhancing the expression of wing-related genes, especially nub. This supports the hypothesis previously proposed by the authors, whereby Br–C might have facilitated the evolution of holometaboly. However, there is no doubt that other factors have also contributed to this evolution.

Expression profiles of an inactive aspartic protease (Bla g 2 allergen) in different tissues and developmental stages of the German cockroach (Blattella germanica).

Tergal glands are found in many insect species and contain constituents such as pheromones, sugars, proteins, and so forth. Preliminary studies have revealed that tergal gland secretions in the German cockroach (Blattella germanica L.) contain the human allergen Bla g 2 (B. germanica allergen 2), an inactive aspartic protease. Although Bla g 2 protein expression has been detected previously in various German cockroach body parts, including male tergal glands, studies that link protein expression in various life stages and tissues with mRNA and protein abundance have not been conducted. Therefore, the goal of this study was to measure the relative abundances of Bla g 2 protein and mRNA in different tissues and life stages of B. germanica using immunoblotting, quantitative PCR, and liquid chromatography‐tandem mass spectrometry (LC‐MS/MS)‐based quantitative profiling. We found that Bla g 2 protein was detected in every sampled tissue, including the male tergal glands. Protein abundance was relatively high in adult males and their tergal glands in comparison to nymphs and virgin females. Similarly, Bla g 2 mRNA transcript levels were also comparatively higher in male tergal glands and adult males. In conclusion, this study provides new information on the relative abundance and distribution of Bla g 2 allergen, a medically significant protein, in different tissues and developmental stages of the German cockroach and lays the foundation for future studies that aim to determine the function of this protein in B. germanica development.

Nephridiophagids (Chytridiomycota) reduce the fitness of their host insects

Nephridiophagids are unicellular fungi (Chytridiomycota), which infect the Malpighian tubules of insects. While most life cycle features are known, the effects of these endobionts on their hosts remain poorly understood. Here, we present results on the influence of an infection of the cockroach Blattella germanica with Nephridiophaga blattellae (Ni = Nephridiophaga-infected) on physical, physiological, and reproductive fitness parameters. Since the gut nematode Blatticola blattae is a further common parasite of B. germanica, we included double infected cockroaches (N + Ni = nematode plus Ni) in selected experiments. Ni individuals had lower fat reserves and showed reduced mobility. The lifespan of adult hosts was only slightly affected in these individuals but significantly shortened when both Nephridiophaga and nematodes were present. Ni as well as N + Ni females produced considerably less offspring than parasite-free (P-free) females. Immune parameters such as the number of hemocytes and phenoloxidase activity were barely changed by Nephridiophaga and/or nematode infections, while the ability to detoxify pesticides decreased. Quantitative proteomics from hemolymph of P-free, Ni, and N + Ni populations revealed clear differences in the expression profiles. For Ni animals, for example, the down-regulation of fatty acid synthases corroborates our finding of reduced fat reserves. Our study clearly shows that an infection with Nephridiophaga (and nematodes) leads to an overall reduced host fitness.

Antimicrobial peptide expression in the cockroach gut during enterobacterial infection is specific and influenced by type III secretion.

ABSTRACTOmnivorous synanthropic cockroaches, such as the German cockroach (Blattella germanica), are reservoirs and vectors of enteric bacterial pathogens. A lifestyle conducive to frequent encounters with high loads of diverse bacteria may have led to the evolution of unique innate immune systems in these insects. The innate immune response of insects relies largely on generalized mechanisms to sense and eliminate foreign microbes. However, analyses of the genomes of common synanthropic cockroaches previously revealed a repertoire of pathogen associated molecular pattern (PAMP) receptors and antimicrobial peptides (AMPs) that is significantly expanded relative to most holometabolous insect models and vectors, supporting the intriguing possibility that cockroaches may encode enhanced recognition within their immune system and may possess an enhanced capacity to fine tune innate immune responses. Investigating how cockroaches respond to infection with enterobacteria provides the opportunity to expand our fundamental knowledge of the regulation of insect innate immunity in a context that is biologically and medically relevant. German cockroaches can harbor both Salmonella enterica serovar Typhimurium and Escherichia coli in their gut without experiencing pathogenesis. The former colonizes the gut and replicates while the latter persists only transiently. We hypothesized that differences in the innate immune response may contribute to or result from the difference in infection dynamics between the two enterobacteria. To test this hypothesis, we used qRT-PCR to analyze expression of five genes encoding representative AMPs (Attacins, Blattellicin, Defensins) in the gut of German cockroaches 1 and 24 h after ingestion of live or heat-killed enterobacteria. We found that robust AMP expression was induced in response to ingestion of a live wild-type strain of S. Typhimurium, but not in response to live E. coli, heat-killed S. Typhimurium, or a live mutant strain of S. Typhimurium lacking type III secretion systems. These results indicate that the cockroach immune system does not respond to stimulation with high levels of ingested bacterial PAMPs such as peptidoglycan. Rather, AMP expression in the gut appears to be induced by active bacterial colonization involving type III secretion. We speculate that this form of regulation may have evolved to prevent over activation of the immune system from frequent ingestion of innocuous, non-colonizing, or non-viable bacteria. While additional work is needed to delineate the molecular mechanisms underlying our observations, our findings provide significant novel insight into the immunological adaptation of cockroaches to life in septic environments as well as the factors that regulate bacterial pathogen transmission by these insects.

MicroRNA let-7 is required for hormonal regulation of metamorphosis in the silkworm, Bombyx mori

The heterochronic microRNA let-7, which was first identified in Caenorhabditis elegans, controls the timing of developmental programs, and let-7 triggers the onset of the juvenile-adult transition in bilaterians. The expression of let-7 is strongly induced during the last larval stage of C. elegans and is highly expressed in the late last instar larvae/nymphs of the fly Drosophila melanogaster and the cockroach Blattella germanica. In the silkworm Bombyx mori, the expression of let-7 remarkably increases in the corpus cardiacum-corpus allatum complex (CC-CA) at the beginning of the last larval instar and is maintained at high levels during this instar. To determine the biological function of let-7 in B. mori, we generated a let-7 knockout line and a transgenic UAS-let-7 line. The let-7 knockout larvae were developmentally arrested in the prepupal stage and became pupal-adult intermediates after apolysis. When let-7 was ubiquitously overexpressed under the transcriptional control of an Actin3-GAL4 driver, developmental timing and growth of larvae were severely impaired in the penultimate (L4) instar, and these larvae underwent precocious metamorphosis from L4. Furthermore, our results showed that reception and signaling of ecdysteroids and juvenile hormones (JHs) normally occurred in the absence of let-7, whereas the biosynthesis of ecdysone and JHs were affected by disruption and overexpression of let-7. Together, the present study demonstrates that let-7 is required for the coordination of the biosynthesis of ecdysone and JH to ensure the developmental transition during the metamorphosis of B. mori.

Population Structure of German Cockroaches (Blattodea: Ectobiidae) in an Urban Environment Based on Single Nucleotide Polymorphisms.

German cockroaches (Blattella germanica L.) harbor and disperse medically important pathogens and are a source of allergens that impact human health and wellbeing. Management of this pest requires an understanding of their distribution and dispersal. In this study, we collected German cockroaches from three apartment buildings in New Jersey, USA. We identified single-nucleotide polymorphisms (SNPs) from DNA extractions using next generation sequencing. We analyzed the SNPs and characterized cockroach population genetic structure using Fst, principal component, phylogenetic, and STRUCTURE analyses. We found significant differences in German cockroach population structure among the buildings. Within buildings, we found variable population structure that may be evidence for multiple colonization events. This study shows that SNPs derived from next generation sequencing provide a powerful tool for analyzing the genetic population structure of these medically important pests.

Genome-wide identification and expression pattern analysis of novel chemosensory genes in the German cockroach Blattella germanica.

The German cockroach Blattella germanica is an important urban insect pest worldwide. In many insects, chemosensation is essential for guiding their behaviors for survival. Although a large number of chemosensory-related genes have been identified in B. germanica, little information on tissue-specific and developmental expression patterns has not been uncovered yet. In this study, we performed transcriptome analysis of different B. germanica tissues to reveal novel chemosensory proteins (CSPs) and sensory neuron membrane proteins (SNMPs). In addition, a phylogenetic tree and gender-specific expression of multiple chemosensory gene families have been analyzed. We identified three CSPs genes (BgerCSP11, BgerCSP12, and BgerCSP13) and five SNMP genes in B. germanica. Tissue-specific expression profiling showed that CSP1, 8, and 9 exhibited significant expression levels in both adult and 5th instar nymph antennae. The results have paved the way for further functional study of the chemosensory mechanism in B. germanica and provided potential insecticide targets.

Of Cockroaches and Symbionts: Recent Advances in the Characterization of the Relationship between Blattella germanica and Its Dual Symbiotic System.

Mutualistic stable symbioses are widespread in all groups of eukaryotes, especially in insects, where symbionts have played an essential role in their evolution. Many insects live in obligate relationship with different ecto- and endosymbiotic bacteria, which are needed to maintain their hosts’ fitness in their natural environment, to the point of even relying on them for survival. The case of cockroaches (Blattodea) is paradigmatic, as both symbiotic systems coexist in the same organism in two separated compartments: an intracellular endosymbiont (Blattabacterium) inside bacteriocytes located in the fat body, and a rich and complex microbiota in the hindgut. The German cockroach Blattella germanica is a good model for the study of symbiotic interactions, as it can be maintained in the laboratory in controlled populations, allowing the perturbations of the two symbiotic systems in order to study the communication and integration of the tripartite organization of the host–endosymbiont–microbiota, and to evaluate the role of symbiotic antimicrobial peptides (AMPs) in host control over their symbionts. The importance of cockroaches as reservoirs and transmission vectors of antibiotic resistance sequences, and their putative interest to search for AMPs to deal with the problem, is also discussed.

Effects of Organic Substrates on Development and Survival of <i>Blattela germanica</i> for Food and Feed in Kenya

Entomophagy has gained popularity as a novel approach to addressing food and nutrition insecurity, particularly in sub-tropical and tropical countries. Cockroaches, for example, have the potential to alleviate nutritional deficiencies, as well as the erratic supply of protein in feeds. They can also be reared on locally available organic substrates. This study investigated growth performance and survival of german cockroach (Blattella germanica) reared on locally available organic matter substrates as treatments; spent brewers’ grain (Treatment A), Treatment B consisting of (40%: wheat bran: 40% spent brewers’ grain: 20% Caridina nilotica), wheat bran (Treatment C), and C. nilotica (Treatment D) and in a completely randomised design (CRD). Each treatment was replicated four times, while in each replicate 20 - 30 nymphs were reared for forty-two days. Feed intake for all the treatments was not significantly different. There was a significant difference (P 0.05) in mature weight, average daily weight gain, live weight, and survival during rearing period. Cockroach performed well on treatment B and recorded highest mature weight, overall performance index, survival was 90.25 mg, 197.35, 96%. The least perfomance was reported in Treatment C; 1.78, 83, 143.17 and 72.5 for average growth rate, survival, overall perfomance and mature weight respectively. B. Based on these findings, a single feed as used in this study was inferior to the composite (Treatment B) in all parameters of growth performance and survival. The composite diet (Treatment B) could be suitable substrate for mass production of B. germanica for feed and food.

New insight into the relationship between Salmonella Typhimurium and the German cockroach suggests active mechanisms of vector-borne transmission

Diarrheal diseases are among the most common illnesses in the world and the bacterium Salmonella enterica serovar Typhimurium is a leading cause of morbidity and mortality from diarrhea globally. The German cockroach (Blattella germanica) frequently harbors and has been linked to human outbreaks of Salmonella, but the mechanisms of vector-borne transmission are not fully clear. Transmission of S. Typhimurium by cockroaches has been previously described as mechanical. Mechanical transmission is a wholly passive process that involves physical transfer of a pathogen from one location or host to another but lacks bacterial replication in the vector and active bacterial processes that promote vector colonization or transmission. Towards the goal of obtaining novel insight into the mechanisms of S. Typhimurium transmission by cockroaches, here we orally provisioned wild type and mutant strains to adult B. germanica and examined several aspects of colonization and shedding. Our results provide evidence of three previously unappreciated phenomena with significant implications. First, we demonstrate that S. Typhimurium undergoes replication at multiple phases during colonization of the cockroach gut. Second, we show the formation of biofilm-like aggregates by S. Typhimurium in the cockroach foregut. Lastly, we identify two mutant strains of S. Typhimurium that are deficient in colonization and shedding relative to isogenic controls, implicating type III secretion and the formation of fimbriae as two processes that are necessary for interaction with the cockroach vector. Together, our data indicate that transmission of S. Typhimurium by cockroaches is not solely mechanical but may resemble biological transmission by other insect vectors that intake human pathogenic bacteria from infected hosts and are subsequently colonized, enabling active dissemination. Thus, these findings suggest that cockroaches and their control may be more important for infection prevention than is currently appreciated. Additional studies to better understand the cycle and biological mechanisms of vector-borne transmission are warranted.

First Report of the L993S Mutation in the Voltage-Gated Sodium Channel in Field Populations of the German Cockroach Blattella germanica.

The long-term and frequent use of pyrethroid insecticides has led to the development of pyrethroid resistance in many insect populations around the world. Specific mutations in the voltage-gated sodium channel (VGSC) have been well documented to be responsible for knockdown resistance (kdr) to pyrethroids and dichlorodiphenyltrichloroethane (DDT) in a variety of arthropods. However, reports regarding naturally occurring kdr mutation in field populations of the German cockroach Blattella germanica (Linnaeus) (Dictyoptera: Blattellidae) in China have remained scarce. In this study, a survey was conducted to detect the presence and frequency of kdr mutations in field populations of B. germanica across Neijiang, Sichuan province of China. In addition to the previously reported L993F mutation, a new amino acid substitution L993S was discovered for the first time. Overall, the classical 993F was the dominant allele with frequencies ranging from 61.8 to 89.6%, while the frequencies of the novel L993S mutation were in the range between 2.5 and 15.0%. Notably, high frequencies (50.0-79.2%) of resistant homozygotes were detected in our samples, indicating high levels of pyrethroid resistance in these B. germanica populations. The results suggest that alternative insecticides with a mode of action different from pyrethroids should be considered in the control of German cockroaches in these regions.

Differential microbial responses to antibiotic treatments by insecticide-resistant and susceptible cockroach strains (Blattella germanica L.)

The German cockroach (Blattella germanica L.) is a major urban pest worldwide and is known for its ability to resist insecticides. Past research has shown that gut bacteria in other insects can metabolize xenobiotics, allowing the host to develop resistance. The research presented here determined differences in gut microbial composition between insecticide-resistant and susceptible German cockroaches and compared microbiome changes with antibiotic treatment. Cockroaches received either control diet or diet plus kanamycin (KAN) to quantify shifts in microbial composition. Additionally, both resistant and susceptible strains were challenged with diets containing the insecticides abamectin and fipronil in the presence and absence of antibiotic. In both strains, KAN treatment reduced feeding, leading to higher doses of abamectin and fipronil being tolerated. However, LC50 resistance ratios between resistant and susceptible strains decreased by half with KAN treatment, suggesting gut bacteria mediate resistance. Next, whole guts were isolated, bacterial DNA extracted, and 16S MiSeq was performed. Unlike most bacterial taxa, Stenotrophomonas increased in abundance in only the kanamycin-treated resistant strain and was the most indicative genus in classifying between control and kanamycin-treated cockroach guts. These findings provide unique insights into how the gut microbiome responds to stress and disturbance, and important new insights into microbiome-mediated insecticide resistance.

Level of Pyrethroid-Resistance Associated with Cytochrome P450 Expression in German Cockroach Blattella germanica (Blattodea: Ectobiidae) in the Field Collected Strains.

Background:Cytochrome P450-dependent monooxygenases are a very important metabolic system involve in insecticide resistance. This study was conducted to find the association between the expression level of cytochrome P450 (CYP450) and permethrin-resistance level among four strains of the German cockroach Blattella germanica (L) (Blattodea: Ectobiidae).Methods:Three field strains of German cockroach with different frequency of exposure to pesticides, and a laboratory susceptible strain were used in the present study. Insecticide susceptibility bioassays were carried out to detect resistance to permethrin. The concentration of CYP450 in each strain was determined using ion-exchange HPLC chromatography. Biochemical assays was performed to analyse CYP450 activities.Results:The resistance ratios (RR) to permethrin among three field strains were 3.29, 4.10 and 6.17-fold comping with the susceptible stain. The CYP450 activity of three field strains was 1.6, 2.4 and 2.7 times higher than in the susceptible stain. The amount of CYP450 per mg of protein was significantly different between the susceptible and the three resistant strains. The resistant cockroaches showed a relatively high expression of CYP450 enzymes. A strong correlation was found between permethrin resistance level and total concentration of CYP450 enzymes.Conclusion:The results of current study show that more frequent usage of a pyrethroid insecticide cause the metabolic insecticide resistance to rise in German cockroach. Therefore, there is a ratio correlation between resistance level and monooxygenases activities in insect. Thus, the control program must be managed according to history of pesticide usage.

Binding of de novo synthesized radiolabeled juvenile hormone (JH III) by JH receptors from the Cuban subterranean termite Prorhinotermes simplex and the German cockroach Blattella germanica

Juvenile hormone (JH) controls insect reproduction and development through an intracellular receptor complex comprising two bHLH-PAS proteins, the JH-binding Methoprene-tolerant (Met) and its partner Taiman (Tai). Many hemimetabolous insects including cockroaches strictly depend on JH for stimulation of vitellogenesis. In termites, the eusocial hemimetabolans, JH also regulates the development of caste polyphenism. Studies addressing the agonist ligand binding to recombinant JH receptors currently include three species belonging to two holometabolous insect orders, but none that would represent any of the hemimetabolous orders. Here, we examined JH receptors in two representatives of Blattodea, the cockroach Blattella germanica and the termite Prorhinotermes simplex. To test the JH-binding capacity of Met proteins from these species, we performed chemical synthesis and tritium labeling of the natural blattodean JH homolog, JH III. Our improved protocol increased the yield and specific activity of [10-3H]JH III relative to formerly available preparations. Met proteins from both species specifically bound [3H]JH III with high affinity, whereas Met variants mutated at a critical position within the ligand-binding domain were incapable of such binding. Furthermore, JH III and the synthetic JH mimic fenoxycarb stimulated dimerization between Met and Tai components of the respective JH receptors of both species. These data present primary evidence for agonist binding by JH receptors in any hemimetabolous species and provide a molecular basis for JH action in cockroaches and termites.

Modulation of fatty acid elongation in cockroaches sustains sexually dimorphic hydrocarbons and female attractiveness.

Insect cuticular hydrocarbons (CHCs) serve as important intersexual signaling chemicals and generally show variation between the sexes, but little is known about the generation of sexually dimorphic hydrocarbons (SDHCs) in insects. In this study, we report the molecular mechanism and biological significance that underlie the generation of SDHC in the German cockroach Blattella germanica. Sexually mature females possess more C29 CHCs, especially the contact sex pheromone precursor 3,11-DimeC29. RNA interference (RNAi) screen against the fatty acid elongase family members combined with heterologous expression of the genes in yeast revealed that both BgElo12 and BgElo24 were involved in hydrocarbon (HC) production, but BgElo24 is of wide catalytic activities and is able to provide substrates for BgElo12, and only the female-enriched BgElo12 is responsible for sustaining female-specific HC profile. Repressing BgElo12 masculinized the female CHC profile, decreased contact sex pheromone level, and consequently reduced the sexual attractiveness of female cockroaches. Moreover, the asymmetric expression of BgElo12 between the sexes is modulated by sex differentiation cascade. Specifically, male-specific BgDsx represses the transcription of BgElo12 in males, while BgTra is able to remove this effect in females. Our study reveals a novel molecular mechanism responsible for the formation of SDHCs and also provide evidences on shaping of the SDHCs by sexual selection, as females use them to generate high levels of contact sex pheromone.

The effectiveness of commercial gel baits against German cockroach Blattella germanica (Linnaeus, 1767) in Indonesia

Gel baits product sold commercially to the public has long been used to control German cockroach populations in urban areas. The effectiveness and resistance level of gel bait products need to be known for the successful control of German cockroaches in the field. However, there have been no reports of resistance in Indonesia. This study used toxicity tests of six gel bait products (MF, CBG, SPB, OTG, BTX, and UBA) against two populations of adult male German cockroaches (VCRU-WHO and RKMN-BKT). Fipronil, boric acid and emamectin benzoate containing gel baits were effective to knock down and kill the German cockroaches. In contrast, gel bait with imidacloprid as an active ingredient was only useful to knock down the German cockroaches. Commercial gel bait product in Indonesia are still useful in controlling German cockroaches’ populations with a time of effectiveness to cause mortality between three to four weeks after being exposed in the field and the level of cockroach resistance to gel bait are susceptible until low resistance.

Laboratory and Field Evaluations of Food-Based Attractants for Monitoring German Cockroaches.

Sticky traps are standard tools for monitoring German cockroaches (Blattella germanica L.); however, because they lack an attractant, their ability to catch cockroaches is by chance and largely dependent upon the location of placement and length of time they are left in place. Currently, highly effective and economical cockroach attractants are not available. Food-based attractants have the potential to be employed in sticky traps and enhance their effectiveness on trapping German cockroaches. We conducted laboratory and field experiments to evaluate the effectiveness of novel food-based attractants. Laboratory results showed that sticky traps containing the following attractants: apple oil, blueberry oil, orange oil, or their combination, fish oil, peanut butter, Roach lure tablet, and bacon extract had significantly higher trap catch compared to those traps that do not contain an attractant. Apple plus blueberry oil mixture and Roach lure tablet enhanced the trap catch by ≥ 103%, and bacon extract enhanced the trap catch by 92% in the field assays. Apple plus blueberry oil mixture and Roach lure tablet were the most promising attractants based on laboratory and field experiments. Apple plus blueberry oil mixture aged for 2 wk was as attractive to German cockroaches as fresh mixture.

SiRNA enrichment in Argonaute 2-depleted Blattella germanica

BackgroundRNA interference (RNAi) is a cellular mechanism used to fight various threats, including transposons, aberrant RNAs, and some types of viruses. This mechanism relies on the detection of dsRNA molecules, which through a pathway involving Dicer-2 (Dcr-2) and Argonaute 2 (AGO2), produces small interfering RNAs (siRNAs) that bind to the complementary RNAs triggering their degradation. MethodsUsing the cockroach Blattella germanica as a model, we examined AGO2 activity by depleting its mRNA using RNAi and analyzing the phenotypes produced. ResultsDepleting AGO2 expression had no remarkable effect on nymphal development or reproduction. dsRNA treatment triggered an immediate and transitory increase in AGO2 expression, independently of Dcr-2 action. In addition, we analyzed the siRNAs generated after injecting a heterologous dsRNA in control and AGO2-depleted animals. The results revealed that obtained siRNAs mapped non-uniformly along the dsRNA sequence. In AGO2-depleted animals, the proportion of 22 nucleotide reads was higher and accumulations of reads appeared in areas less well-represented in the controls. We also detected a preference for cytosine as the first nucleotide in controls that was significantly attenuated in AGO2-depleted individuals. Conclusions/general significanceThe siRNAs produced from a dsRNA mapped heterogeneously along the length of the dsRNA and this arrangement depends on the dsRNA sequence. AGO2 exerts its role as nuclease on the siRNA duplexes independently of its action on the corresponding mRNA. This study sheds light on an extremely useful process for reverse genetics in laboratories, in addition to the design of more effective, specific, and eco-friendly pest-control strategies.

First report of Lophomonas spp. in German cockroaches (Blattella germanica) trapped in hospitals, northern Iran.

Cockroaches are considered to be important reservoirs for protozoan and helminth parasites,particularly an emerging Lophomonas spp., protozoan parasite. German cockroaches, Blattella germanica (B. germanica), are domestic pest distributed worldwide. However, little information is available regarding the burden of Lophomonas spp. infestation among domestic cockroach population worldwide. The present study investigated parasitic agents, emphasizing Lophomonas spp. infestation among B. germanica in northern Iran. During spring and summer 2018, a total of 496 adults B. germanica (265 males and 231 females) were trapped by hand and glass traps from the teaching hospitals in Mazandaran, Sari, northern Iran. All cockroaches were identified using taxonomic keys. Each cockroaches was anesthetized with chloroform and the alimentary tract removed. Each section was smeared onto a glass slide. All the smears were examined under light microscopy to determine the presence of the parasites. Ten genera of parasites were identified from the trapped B. germanica; overall, 233 (47%) were found to be positive for helminths (n = 135) and protozoa (n = 98). The helminths included Aspiculuris tetraptera egg (67/233, 28.7%), Oxiuros sp. (39/233; 16.7%), Dentostomella translucida (17/233; 7.3%), and nematodes Ova (12/233; 5.2%). The protozoan parasites identified Gregarina sp. (61/233; 26.2%), Entamoeba sp. (19/233; 8.2%), Blastocystis sp. (5/233; 2.1%), Lophomonas (4/233; 1.7%), Nyctotherus sp. (4/233; 1.7%), unclassified flagella (5/233; 2.1%). Lophomonas spp. was detected within the gut of B. germanica, for the first time, in Iran. It has previously been identified in the gut of cockroaches a long time ago, however, this is the first attempt to detect Lophomonas in cockroaches housing in hospitals. Thereby, it possibly presenting pose a serious respiratory infection risk for patients and health care personnel.