The resistance dynamics of Blattella germanica in Yangpu District of Shanghai from 2014 to 2020 / 上海预防医学

Objective To investigate the resistance dynamics of Blattella germanica to commonly used insecticides in Yangpu District of Shanghai during 2014‒2020， and to provide the basis for the rational use of insecticides against cockroaches. Methods Blattella germanica was captured in two farmer’s markets in 2014， 2017 and 2020. The residual film method recommended by WHO was used to test resistance of Blattella germanica to the insecticides. Results In 2014， 2017 and 2020， the resistance ratio of Blattella germanica in Yangpu District was 6.96， 4.39， 7.26 to beta-cypermethrin， 10.41， 6.71， 5.24 to permethrin， and 2.28， 3.16， 1.01 to propoxur. Conclusion The resistance of Blattella germanica to permethrin and propoxur decreases， and the resistance to beta-cypermethrin keeps at low level. Comprehensive management and rotation of insecticides should be implemented to delay the increase of resistance.

Laboratory study on chain-killing efficacy of four gel baits / 上海预防医学

Objective To study the first-time killing efficacy and the chain-killing efficacy of four gel baits against Blattella germanica: 1% chlorpyrifos, 0.05% fipronil, 2.15% imidacloprid, and 0.5% dinotefuran and provide a basis for drug selection in controlling Blattella germanica. Methods Laboratory killing efficacy test was conducted according to the national standard GB/T 13917.7-2009 and the chain-killing efficacy test was conducted for three rounds.The first round of chain efficacy test was conducted by feeding the cockroaches killed in the laboratory efficacy test, and each next round by feeding the cockroaches killed in the last round.Median lethal time (LT50), 95% confidence limit, and toxicological regression equation of each test were calculated by software DPS V9.01. Results The LT50 of the efficacy test with 1% chlorpyrifos gel bait was 0.745 5 (0.603 4-0.890 3) d.The LT50 of the first, second and third chain experiments increased by 3.30, 2.18 and 2.76 times, respectively.The LT50 of the efficacy test with 0.05% fipronil gel bait was 0.846 5(0.464 7-1.228 0)d, and increased by 5.42, 2.09 and 1.48 times, respectively, in the first, second and third chain experiments.The LT50 of the efficacy test with 2.15% imidacloprid gel bait was 3.192 1(2.865 0-3.506 0)d, and increased by 1.13, 1.65 and 1.15 times, respectively in the first, second and third chain experiments.The LT50 of the efficacy test with 0.5% dinotefuran gel bait was 0.997 1(0.805 8-1.191 6) d, and increased by 3.85, 1.37 and 1.78 times, respectively in the first, second and third chain experiments. Conclusion In the laboratory killing efficacy test, 1% chlorpyrifos, 0.05% fipronil, and 0.5% dinotefuran gel baits are better than 2.15% imidacloprid gel bait.In the chain-killing efficacy test, 2.15% imidacloprid and 0.5% dinotefuran gel baits are better than 1% chlorpyrifos and 0.05% fipronil gel baits.Based on our results, we recommend the use of 0.5% dinotefuran gel bait for comprehensive and sustained killing effect.

Synergists action of piperonyl butoxide and S,S,S-tributyl phosphorotrithioate on toxicity of carbamate insecticides against Blattella germanica

OBJECTIVE@#To determine the synergists action of piperonyl butoxide (PBO) and S,S,S-tributyl phosphorotrithioate (DEF) on toxicity of carbamate insecticides against Blattella germanica in Tehran city.@*METHODS@#In the current study, German cockroach strains were collected from several hospitals and dormitories in Tehran. At the beginning, different concentrations of bendiocarb and carbaryl (insecticides belong to carbamate group) were determined by surface contact on a susceptible strain. Then, the level of susceptibility and type of resistance mechanisms in the collected strains from contaminated sites to the aforementioned insecticides were studied by using PBO and DEF synergists with different insecticide ratios to synergist (1:0, 1:1, 1:2, 1:3).@*RESULTS@#The DEF synergist along with bendiocarb and carbaryl completely eliminated the resistance in all strains but PBO did not completely eliminate the resistance in the strains of Mofid, Alvand, Valiasr hospitals and Shariati dormitory. Generally, the impact of DEF was observed in the removing resistance more than PBO.@*CONCLUSIONS@#In most of these strains, resistance to bendiocarb and carbaryl is completely eliminated by DEF, showing a very high role of estraze enzymes in resistance to bendiocarb and carbaryl. But in most strains PBO does not remove the resistance because other mechanisms, such as reduced cuticle penetration and insensitivity to the acetylcholine esterase enzyme, may be involved.

Synergists action of piperonyl butoxide and S,S,S-tributyl phosphorotrithioate on toxicity of carbamate insecticides against Blattella germanica

Objective To determine the synergists action of piperonyl butoxide (PBO) and S,S,S-tributyl phosphorotrithioate (DEF) on toxicity of carbamate insecticides against Blattella germanica in Tehran city. Methods In the current study, German cockroach strains were collected from several hospitals and dormitories in Tehran. At the beginning, different concentrations of bendiocarb and carbaryl (insecticides belong to carbamate group) were determined by surface contact on a susceptible strain. Then, the level of susceptibility and type of resistance mechanisms in the collected strains from contaminated sites to the aforementioned insecticides were studied by using PBO and DEF synergists with different insecticide ratios to synergist (1:0, 1:1, 1:2, 1:3). Results The DEF synergist along with bendiocarb and carbaryl completely eliminated the resistance in all strains but PBO did not completely eliminate the resistance in the strains of Mofid, Alvand, Valiasr hospitals and Shariati dormitory. Generally, the impact of DEF was observed in the removing resistance more than PBO. Conclusions In most of these strains, resistance to bendiocarb and carbaryl is completely eliminated by DEF, showing a very high role of estraze enzymes in resistance to bendiocarb and carbaryl. But in most strains PBO does not remove the resistance because other mechanisms, such as reduced cuticle penetration and insensitivity to the acetylcholine esterase enzyme, may be involved.

Cost-effectiveness of integrated pest management compared with insecticidal spraying against the German cockroach in apartment buildings

This study assessed the cost and effectiveness of an integrated pest management (IPM) program using hydramethylnon gel baits compared with conventional spraying for controlling the German cockroach, Blattella germanica (L.) (Blattodea: Blattellidae), in two residential buildings in Yasuj, Iran. The IPM approach was based on educational programs using pamphlets, posters and lectures, sanitation using vacuuming and application of hydramethylnon gel baits. Conventional approach used cypermethrin (10 percent EC) on baseboard and cracks-and-crevices. Sticky traps were used as tools for monitoring cockroach population densities. The IPM approach reduced (943 percent) the rate of insecticide application compared to the conventional spray. Cockroach populations in the IPM treatment were significantly reduced from an average of 12.2 ± 3.01 cockroaches per unit before treatment to zero cockroach per unit by week four and thereafter. Cockroach populations in the conventional spray treatment were reduced from an average of 11.5 ± 4.43 cockroaches per unit before treatment to an average of 3.4 ± 0.99 cockroach per unit after 11 weeks of post treatment. The IPM treatment improved 100 percent of infested units compared to 78 percent for spray treatment to obtain a clean level of infestation (< 1cockroach per trap per unit). The results suggest that the intervention by IPM using hydramethylnon gel baits significantly reduced cockroach infestation compared to cypermethrin spray throughout the 11 weeks of post-treatment period. However, within the study period, the IPM system involving gel baits, educational program and sanitation was 363.2 percent more expensive than the conventional method.

A revisit to cockroach allergens.

Among cockroaches (CR) that live in people’shomes, two species, i.e., German CR (Blattella germanica) and American CR (Periplaneta americana) predominate in temperate and tropical areas, respectively. CR is an important source of inhalant indoor allergens that sensitize atopic subjects to (localized) type I hypersensitivity or atopy including allergic rhinitis and atopic asthma. In Thailand the predominant CR species is P. americana. CR allergens are found throughout CR infested houses; the number found in kitchens correlates with the degree of CR infestation while sensitization and reactivation of the allergic morbidity are likely to occur in the living room and bedroom. Levels of the CR allergens in homes of CR allergic Thais, measured by using locally made quantification test kits, revealed that the highest levels occur in dust samples collected from the wooden houses of urban slums and in the cool and dry season. CR allergens are proteins that may be derived from any anatomical part of the insect at any developmental stage. The allergens may be also from CR secretions, excretions, body washes or frass. The proteins may be the insect structural proteins, enzymes or hormones. They may exist as dimers/multimers and/or in different isoforms. Exposure to CR allergens in infancy leads to allergic morbidity later in life. Clinical symptoms of CR allergy are usually more severe and prolonged than those caused by other indoor allergens. The mechanisms of acute and chronic airway inflammation and airway hyper-responsiveness (AHR) have been addressed including specific IgE- and non-IgEmediated mechanisms, i.e., role of proteaseactivated receptor-2 (PAR2). Participation of various allergen activated-CD4+ T cells of different sublineages, i.e., Th2, Th17, Th22, Th9, Th25, Tregs/Th3 as well as invariant NKT cells, in asthma pathogenesis have been mentioned. The diagnosis of CR allergy and the allergy intervention by CR population control are also discussed.

Enzymatic Activities of Allergen Extracts from Three Species of Dust Mites and Cockroaches Commonly Found in Korean Home

Allergen extracts from dust mites and cockroaches commonly found in Korean homes were used to evaluate their enzymatic activity as they are believed to influence allergenicity. Allergen extracts were prepared from 3 dust mite species (Dermatophagoides farinae, D. pteronyssinus, and Tyrophagus putrescentiae) and 3 cockroach species (Blattella germanica, Periplaneta americana, and P. fuliginosa) maintained in the Korea National Arthropods of Medical Importance Resource Bank. Proteins were extracted in PBS after homogenization using liquid nitrogen. The activities of various enzymes were investigated using the API Zym system. No significant difference in phosphatase, lipase, or glycosidase activity was observed among the 6 allergen extracts, but much difference was observed in protease activity. Protease activity was assessed in more detail by gelatin zymography and the EnzChek assay. Extract from T. putrescentiae showed the highest protease activity, followed by those of the cockroach extracts. Extracts from D. farinae and D. pteronyssinus showed only weak protease activity. Gelatinolytic activity was detected mainly in a 30-kDa protein in D. farinae, a 28-kDa protein in D. pteronyssinus, a > 26-kDa protein in T. putrescentiae, a > 20-kDa protein in B. germanica, and a > 23-kDa protein in P. americana and P. fuliginosa. The information on various enzymatic activities obtained in this study may be useful for future studies. In particular, the strong protease activity found in cockroach extracts could contribute to sensitization to cockroach allergens, which is known to be associated with the development of asthma.

Resistencia a deltametrina de cepas de Blattella germanica (Dictyoptera: Blattellidae) en la ciudad de Cali, Colombia / Deltamethrin resistance in Blattella germanica (Dyctioptera: Blattellidae) strains from Cali city, Colombia

estatus de plaga se debe a su gran capacidad de infestación y al desarrollo de resistencia a insecticidas en corto plazo. Como no se han realizado estudios locales previos es necesario determinar el estado de la resistencia a diferentes insecticidas, datos que servirán de referencia para establecer un sistema de vigilancia que mejorará las medidas de control. OBJETIVO: establecer el estado de la resistencia al piretroide deltametrina en poblaciones de B. germanica de la ciudad de Cali (Colombia). MÉTODOS: se colectaron 5 cepas de B. germanica en la ciudad de Cali, en lugares con altos niveles de infestación. Se evaluaron adultos machos y hembras de esta especie expuestos al piretroide deltametrina mediante bioensayos con botellas. RESULTADOS: en machos y hembras de B. germanica de la ciudad de Cali, se determinó la CK50 a deltametrina. Los machos fueron más susceptibles que las hembras. Dos de las cepas evaluadas presentaron un elevado factor de resistencia (Club deportivo: FR= 22 para hembras y 13 para machos; Plaza de mercado 2: FR= 14,33 para hembras y 13,5 para machos) y corresponden a áreas con una historia de mayor frecuencia de aplicación de insecticidas. La cepa más susceptible (Hospital: CK50= 0,03 mg/mL y 0,02 mg/mL para hembras y machos, respectivamente) no ha estado sometida a una presión de selección constante con insecticidas, debido a los riesgos que implican estas aplicaciones en lugares donde se encuentran pacientes con tratamientos médicos delicados. CONCLUSIONES: estos resultados constituyen el primer registro de resistencia a deltametrina en poblaciones de B. germanica en Cali (Colombia). En las poblaciones susceptibles se puede continuar la aplicación de deltametrina dentro de un programa de vigilancia y control permanente.

INTRODUCTION: Blattella germanica is one of the most important pests worldwide. The pest status is due to their great infestation capacity and ability to develop relative short-term insecticide resistance. Since local studies about resistance on B. germanica to any insecticide have not been made, it is necessary to find out the state of resistance of this species to insecticides. The collected data will allow establishing an effective monitoring system that will improve some control measures already in place. OBJECTIVE: to ascertain the level of resistance to deltamethrin in some populations of B. germanica from Cali (Colombia). METHODS: five different strains of B. germanica were collected in Cali (Colombia), in areas with high levels of infestation. Adult males and females were exposed and tested to deltamethrin using bottled bioassays. RESULTS: KC50 to pyrethroid deltametrhin was determined in adult males and females of B. germanica from Cali city. Males were more susceptible than females. Two evaluated strains showed a high resistance factor (Sports Club: RT=22 for females and 13 for males; Market 2: RT=14.33 for females and 13.50 for males) representing areas with history of highly frequent insecticide application. The most susceptible strain (Hospital: KC50=0.03 mg/mL and 0.02 mg/mL for females and males respectively) has not been submitted to constant selection pressure with insecticides because of the likely risks of this type of application for human health in places where critically-ill patients are treated.. CONCLUSIONS: this study contributed the first evidence of deltamethrin resistance in populations of B. germanica from Cali (Colombia). It is possible to carry on using deltamethrin in susceptible populations within an ongoing monitoring and control program.

IgE Binding Reactivity of Peptide Fragments of Bla g 4, a Major German Cockroach Allergen

Cockroaches have been recognized as a major cause of asthma. Bla g 4 is one of the most important German cockroach allergens. The aim of this study is to investigate IgE reactivity to the recombinant Bla g 4 (rBla g 4) in the sera of allergic patients and identify linear IgE binding epitope. For protein expression, full-length Bla g 4 (EF202172) was divided into 5 overlapping peptide fragments (E1: aa 1-100, E2: aa 34-77, E3: aa 74-117, E4: aa 114-156, and E5: aa 153-182). The full-length and 5 peptide fragments of Bla g 4 was generated by PCR and over-expressed in E. coli BL21 (DE3). The IgE binding reactivities of the full-length and peptide fragments were measured by ELISA using 32 serum samples of cockroach allergy. The sera of 8 patients (25%) reacted with rBla g 4. Four sera (100%) showed IgE-binding reactivity to full-length and peptide fragment 4, and 2 sera (50%) reacted with peptide fragment 2. One (20%) serum reacted with peptide fragment 3. The results of ELISA using overlapping recombinant fragments indicated that the epitope region was located at amino acid sequences 34-73 and 78-113, and major IgE epitope of Bla g 4 was located at amino acid sequences 118-152 of C-terminal. B-cell epitope analysis of German cockroach allergen Bla g 4 could contribute to the strategic development of more specific and potentially efficacious immunotherapy.

Cloning of a Family of Serine Protease Genes Expressed in German Cockroach(Blattella germanica) and Possibility as Allergens / 대한이비인후과학회지

BACKGROUND AND OBJECTIVES: The serine protease allergens have been known as one of the most important inhalent allergens of insects as well as house dust mites. There is no known serine protease allergen of German cockroach, which is a well-known etiological agent of allergic diseases. We identified the serine protease genes of cockroach, Blattella germanica, and examined its possibility as an allergen. MATERIALS AND METHOD: We have isolated three partial genes, Bg3, Bg5 and Bg6, of serine protease from the Blattella germanica cDNA library using the degenerate oligonucleotide PCR primers specific for the conserved regions. RESULTS: The genes, Bg3, Bg5 and Bg6, were composed of 396 bp 131 amino acids, 513 bp 170 amino acids, and 410 bp 136 amino acids, respectively. Northern hybridization analysis indicated that the size of Bg5 and Bg6 transcripts was approximately 0.7 kb. The gene, Bg5, was also identified as a multiple gene by the Southern blot analysis. The gene, Bg3, showed significant homology to trypsin-like serine protease of various insects. The deduced amino acids sequence of the gene Bg5 was matched to the mite group III allergen as well as various species of insect serine protease sequences. The gene Bg6 also showed high homology to the amino acids sequences of insect serine proteases and mite group III allergens. Three Blattella germanica serine protease gene fragments revealed close genetic relationships with Dermatophagoides farinae group III allergen in the phylogenetic analysis. CONCLUSION: The high homology and close phylogenetic relationship raise the possibility of three serine protease genes as being an allergen of German cockroach.

Reactivity of German Cockroach Allergen, Bla g 2, Peptide Fragments to IgE Antibodies in Patients' Sera

Bla g 2 is a cockroach allergen of great importance. This study was conducted to identify IgE-binding epitope(s) of Bla g 2 using the recombinant protein technique. Approximately 50% of tested sera showed IgE reactivity to Pichiaexpressed Bla g 2 (PrBla g 2) and E. coli-expressed Bla g 2 (ErBla g 2). Only 5.3% of serum samples showed stronger reactivity to PrBla g 2 than ErBla g 2, indicating that serum was reactive to conformational or carbohydrate epitopes. The full-length and 5 peptide fragments of Bla g 2 were produced in E. coli. All fragments showed IgE-binding activity to the cockroach-allergy patients' sera. Specifically, peptide fragments of amino acid residue 1-75 and 146-225 appeared to be important for IgE-binding. The information about the IgE-binding epitope of Bla g 2 can aid in the diagnosis and treatment for cockroach allergies.

Effectiveness of education for control of house dust mites and cockroaches in Seoul, Korea

We evaluated the efficacy of health education in reducing indoor arthropod allergens in Seoul. The mite control measures comprised the use of mite-proof mattress and pillow coverings, regular washing of potentially infested materials, maintenance of a low humidity, removal of carpets, and frequent vacuum cleaning. Cockroach control measures included trapping, application of insecticides, and protecting food. Of 201 homes enrolled in October 1999, 63 volunteers were included in a 2-year follow-up survey between April 2000 and January 2002. Before intervention, the density of mites/g of dust varied greatly; 27.1/g in children's bedding, 20/g in adult bedding, 7.2/g on the floors of children's bedrooms, 6.8/g in sofas, 5.9/g on the floors of adult's bedrooms, 3.9/g on living room floors, 3.7/g in carpets, and 1.9 mites/g on kitchen floors. The predominant mite species and house percentages infested were; Dermatophagoides farinae 93%, D. pteronyssinus 9%, and Tyrophagus putrescentiae 8%. Comparing 1999 and 2001 infestations, before and after 25 mo of education, mite abundance was reduced by 98%, from 23.7 to 0.57 mites/g of dust. In 1999, cockroaches were detected in 62% homes: 36% Blattella germanica and 35% Periplaneta spp., including 9% double infestations of B. germanica and P. americana. Following intervention, cockroach infestation rates decreased to 22% of houses in 2000 and 23% in 2001. We conclude that continuous and repetitive health education resulted in the effective control of domestic arthropods.

Expressed sequence tags analysis of Blattella germanica

Four hundred and sixty five randomly selected clones from a cDNA library of Blattella germanica were partially sequenced and searched using BLAST as a means of analyzing the transcribed sequences of its genome. A total of 363 expressed sequence tags (ESTs) were generated from 465 clones after editing and trimming the vector and ambiguous sequences. About 42% (154/363) of these clones showed significant homology with other data base registered genes. These new B. germanica genes constituted a broad range of transcripts distributed among ribosomal proteins, energy metabolism, allergens, proteases, protease inhibitors, enzymes, translation, cell signaling pathways, and proteins of unknown function. Eighty clones were not well-matched by database searches, and these represent new B. germanica-specific ESTs. Some genes which drew our attention are discussed. The information obtained increases our understanding of the B. germanica genome.

Cloning and expression of trypsin-encoding cDNA from Blattella germanica and its possibility as an allergen

In this study, the trypsin gene (bgtryp-1) from the German cockroach, Blattella germanica, was cloned via the immunoscreening of patients with allergies to cockroaches. Nucleotide sequence analysis predicted an 863 bp open reading frame which encodes for 257 amino acids. The deduced amino acid sequence exhibited 42-57% homology with the serine protease from dust mites, and consisted of a conserved catalytic domain (GDSGGPLV). bgtryp-1 was determined by both Northern and Southern analysis to be a 0.9 kb, single-copy gene. SDS-PAGE and Western blotting analyses of the recombinant protein (Bgtryp-1) over-expressed in Escherichia coli revealed that the molecular mass of the expressed protein was 35 kDa, and the expressed protein was capable of reacting with the sera of cockroach allergy patients. We also discussed the possibility that trypsin excreted by the digestive system of the German cockroach not only functions as an allergen, but also may perform a vital role in the activation of PAR-2.

Expression of Allergen Bla g 2 from Blattella germanica in Pichia pastoris / 中国寄生虫学与寄生虫病杂志

Objective To express the major allergen of Blattella germanica (Bla g 2) in Pichia pastoris and obtain the soluble protein. Methods The known Bla g 2 gene was used to design the primers which had the restriction enzyme sites. PCR method was applied to obtain the Bla g 2 gene. The gene fragment was then cut and ligated with the Pichia expression vector pGAPZaA, resulting in a recombinant plasmid pGAPZaA-Bla g 2. The linearized pGAPZaA-Bla g 2 was transformed into Pichia pastoris GS115 through electroporation, then screened to positive transformants, and the protein was expressed in YPD medium. Purification of the recombinant protein was achieved by metal (Ni2+) chelating affinity chromatography and Western-blotting assay indicated its IgE binding capacity. Results With the expressed reeombinanl protein, SDS-PAGE showed the presence of the product in the supernatant of the culture with Mr 45 000. After 3 days culture, the recombinant protein occupied 50% of the total proteins in the supernatant. The recombinant protein was purified and Western-blot demonstrated an adequate IgE binding capacity of the product. Conclusion A recombinant protein of Bla g 2 has been obtained, which is soluble in the supernatant and therefore can avoid a process of denaturalization and renaturation of the recombinant.