Laboratory Insecticide Efficacy Trials of Lethal Harborages for Control of the Common Bed Bug, Cimex lectularius (Hemiptera: Cimicidae).

Over the past two decades, there has been a worldwide resurgence in the bed bugs Cimex lectularius L. and Cimex hemipterus (F.). This is primarily due to insecticide resistance, making bed bug management and eradication challenging and expensive. To address the need for more affordable control solutions, "lethal harborages" were explored. Cardboard squares were treated using insecticidal dusts at different dosage levels, including silica dioxide, diatomaceous earth, deltamethrin, permethrin, and fipronil. Two strains of C. lectularius, one susceptible and one resistant, were allowed to enter the treated harborages, and mortality rates were recorded daily. The silica dioxide products proved to be the most efficacious, consistently achieving 100% mortality between 14-17 d at the highest dose. An artificial environment trial using the "new ChinChex®" formulation of silica dioxide resulted in the complete elimination of bed bugs in the treated harborages within 21 d. These findings suggest that lethal harborages, especially those impregnated with silica dioxide, offer a cost-effective solution that could be incorporated into broader integrated bed bug management strategies. This approach may help alleviate the burden of bed bug infestations in economically disadvantaged communities.

Long-term clinical outcomes of non-melanoma skin cancer patients treated with electronic brachytherapy.

Purpose: High-dose-rate electronic brachytherapy (eBx) is a non-surgical treatment option for non-melanoma skin cancer (NMSC) patients. This study assessed long-term effectiveness and safety of eBx for the treatment of NMSC. Material and methods: A chart review was conducted to identify subjects who had five or more years since their last eBx treatment fraction. Subjects meeting these criteria were contacted to determine their interest in participating in a long-term follow-up study. Those who agreed, underwent a follow-up visit where consent was obtained, and their lesions were clinically assessed for recurrence and long-term skin toxicities. History and demographic data were retrospectively collected, and treatment method was verified. Results: 183 subjects with 185 lesions were enrolled into this study at four dermatology centers in two practices in California. Three subjects in the analysis were less than 5 years from the last treatment to follow-up visit. All lesions were stage 1 basal cell carcinoma, squamous cell carcinoma, or squamous cell carcinoma in situ. Recurrence rate for the 183 subjects was 1.1%. Long-term skin toxicities were reported in 70.0% of the subjects. Hypopigmentation grade 1 was observed in 65.9% of the lesions, telangiectasia grade 1 was seen in 22.2%, scarring grade 1 in two subjects (1.1%), hyperpigmentation grade 1 in two subjects (1.1%), and induration grade 2 in one patient (0.5%). The induration grade 2 was located on the upper back and did not limit instrumental activities of daily living (ADLs). Conclusions: Electronic brachytherapy for the treatment of non-melanoma skin cancer is safe and effective, showing excellent long-term 98.9% local control through a median follow-up of 7.6 years (n = 183), with minimal long-term toxicities.

Performance of Pyrethroid-Neonicotinoid Mixture Formulations Against Field-Collected Strains of the Tropical Bed Bug (Hemiptera: Cimicidae) on Different Substrates.

The residual performance of two pyrethroid-neonicotinoid mixture formulations: Temprid SC (10.5% beta-cyfluthrin and 21% imidacloprid) and Tandem (3.5% lambda-cyhalothrin and 11.6% thiamethoxam) on two substrates (glass and filter paper) against eight pyrethroid-resistant strains (BM-MY, BP-MY, CH-MY, GL-MY, KL-MY, SAJ-MY, TT-MY, and QLD-AU) of the tropical bed bug, Cimex hemipterus (F.) (Hemiptera: Cimicidae) collected from Malaysia, and Australia were evaluated. The aging effect of treatment residues on glass was also investigated. A susceptible C. lectularius L. strain (Monheim) was used for comparison. Temprid SC showed varying levels of performance against all C. hemipterus strains: TT-MY (PR50 = 6.5-fold, high performance), BM-MY, GL-MY, SAJ-MY, and QLD-AU (12.8-21.6-fold, moderate performance), BP-MY, and KL-MY (48.2-49-fold, poor performance), CH-MY (128.2-fold, very poor performance). On the other hand, Tandem displayed high performance against all C. hemipterus strains (1.8-8.3-fold). Tandem caused faster mortality than Temprid SC for all strains. Temprid SC and Tandem residues killed C. hemipterus significantly faster on glass than filter paper. Compared with fresh residues, the efficacy of Temprid SC residues significantly declined after one week of aging, while the effectiveness of Tandem residues declined after two weeks of aging. Further investigations using the topical assay method with a diagnostic dose of imidacloprid found two strains (CH-MY and GL-MY) resistant to imidacloprid. The six other strains (BM-MY, BP-MY, KL-MY, SAJ-MY, TT-MY, and QLD-AU) were susceptible.

The Efficacy of a Pyrethroid-impregnated Mattress Liner on Multiple International Strains of Cimex lectularius (Hemiptera: Cimicidae) and Cimex hemipterus (Hemiptera: Cimicidae).

Modern bed bugs are resistant to multiple insecticide classes, particularly the pyrethroids. The efficacy of pyrethroid-impregnated mattress liners marketed for bed bug management has been variable. This study evaluated the efficacy of a permethrin-impregnated mattress liner, ActiveGuard, against 24 bed bug strains, consisting of both Cimex hemipterus (F.) and Cimex lectularius L. A 'mat assay', employing an allethrin-impregnated mat, was used to establish the pyrethroid resistance profile of all strains. Three experiments were conducted to evaluate the effect of ActiveGuard exposure on bed bug knockdown: 1) exposing the bed bugs continuously on the liner for up to 24 d, 2) holding the bed bugs on the liner for either 4 or 6 h, and 3) placing a noninsecticide treated fabric above the liner with the bed bugs held continuously on top. Our results indicated that all modern strains (collected within the last 15 years during the current resurgence) were pyrethroid-resistant, although the magnitude of resistance was highly variable between strains. In the continuous exposure study, an incomplete knockdown was recorded for most modern bed bug strains, with some having no knockdown even up to 7 d of constant exposure. In the 4 or 6 h exposure study, the level of knockdown was reduced even further, and very few bed bugs were knocked down in the double fabric study. The results of this study indicate that pyrethroid-impregnated mattress liners are not likely to be effective in the management of most modern bed bug infestations involving either C. hemipterus or C. lectularius.

Historical and Contemporary Control Options Against Bed Bugs, Cimex spp.

Bed bugs (Hemiptera: Cimicidae) are an important group of obligate hematophagous urban insect pests. The global resurgence of bed bugs, involving the common bed bug, Cimex lectularius L., and the tropical bed bug, Cimex hemipterus (F.), over the past two decades is believed to be primarily due to the development of insecticide resistance, along with global travel and poor pest management, which have contributed to their spread. This review examines and synthesizes the literature on bed bug origins and their global spread and the literature on historical and contemporary control options. This includes bed bug prevention, detection and monitoring, nonchemical and chemical control methodologies (and their limitations), and potential future control options. Future research needs are highlighted, especially the factors behind the modern resurgence, the necessity of identifying differences between the two bed bug species relevant to control, and the need to improve insecticide test protocols and management strategies.

Rats and the city: Implications of urbanization on zoonotic disease risk in Southeast Asia.

Urbanization is rapidly transforming much of Southeast Asia, altering the structure and function of the landscape, as well as the frequency and intensity of the interactions between people, animals, and the environment. In this study, we explored the impact of urbanization on zoonotic disease risk by simultaneously characterizing changes in the ecology of animal reservoirs (rodents), ectoparasite vectors (ticks), and pathogens across a gradient of urbanization in Kuching, a city in Malaysian Borneo. We sampled 863 rodents across rural, developing, and urban locations and found that rodent species diversity decreased with increasing urbanization-from 10 species in the rural location to 4 in the rural location. Notably, two species appeared to thrive in urban areas, as follows: the invasive urban exploiter Rattus rattus (n = 375) and the native urban adapter Sundamys muelleri (n = 331). R. rattus was strongly associated with built infrastructure across the gradient and carried a high diversity of pathogens, including multihost zoonoses capable of environmental transmission (e.g., Leptospira spp.). In contrast, S. muelleri was restricted to green patches where it was found at high densities and was strongly associated with the presence of ticks, including the medically important genera Amblyomma, Haemaphysalis, and Ixodes. Our analyses reveal that zoonotic disease risk is elevated and heterogeneously distributed in urban environments and highlight the potential for targeted risk reduction through pest management and public health messaging.

Emergence of Japanese encephalitis in Australia: a diagnostic perspective.

The unprecedented emergence of Japanese encephalitis (JE) in mainland Australia represents an outbreak of high clinical and public health significance. JE is a zoonosis spread by mosquitoes and is one of the most important causes of endemic viral encephalitis in South-East Asia and the Indian subcontinent. While occasional cases of human Japanese encephalitis virus (JEV) infection have occurred in far north Australia, its detection in pigs and the substantial number of locally acquired human cases across multiple jurisdictions in early 2022 prompted the declaration of this outbreak as a Communicable Disease Incident of National Significance. Laboratory testing for JEV is complex, and most cases are diagnosed by serology, for which interpretation is difficult. This review provides a comprehensive outline of currently available methods for JEV diagnosis including serology, nucleic acid amplification testing, virus isolation, sequencing and metagenomics. The relative advantages and disadvantages of the diagnostic tests are presented, as well as their value in clinical and public health contexts. This review also explores the role of mosquito, veterinary and human surveillance as part of the laboratory response to JEV. As JEV may become endemic in Australia, a collaborative and coordinated One Health approach involving animal, human and environmental health is required for optimal disease response and control.

Weather extremes associated with increased Ross River virus and Barmah Forest virus notifications in NSW: learnings for public health response.

OBJECTIVE: To examine the sequence of environmental and entomological events prior to a substantial increase in Ross River virus (RRV) and Barmah Forest virus (BFV) notifications with a view to informing future public health response. METHODS: Rainfall, tidal, mosquito and human arboviral notification data were analysed to determine the temporality of events. RESULTS: Following two extremely dry years, there was a substantial increase in the abundance of mosquitoes along coastal New South Wales (NSW) two weeks after a significant rainfall event and high tides in February 2020. Subsequently, RRV and BFV notifications in north east NSW began to increase eight and nine weeks respectively after the high rainfall, with RRV notifications peaking 12 weeks after the high rainfall. CONCLUSIONS: Mosquito bite avoidance messaging should be instigated within two weeks of high summer rainfall, especially after an extended dry period. IMPLICATIONS FOR PUBLIC HEALTH: Intense summertime rain events, which are expected to increase in frequency in south-east Australia with climate change, can lead to significant increases in arboviral disease. These events need to be recognised by public health practitioners to facilitate timely public health response. This has taken on added importance since the emergence of Japanese encephalitis virus in southeastern Australia in 2022.

Morphometrics of the Tropical Bed Bug (Hemiptera: Cimicidae) From Cape Coast, Ghana.

Bed bugs, Cimex lectularius (L.) (Hemiptera: Cimicidae) and Cimex hemipterus (F.), have become established worldwide in recent years largely due to the development of insecticide resistance. However, limited attention has been given to ongoing morphological and macroevolutionary changes within the species and their populations, which could have implications for their control. Here, we evaluated whether bed bugs of the species C. hemipterus inhabiting different communities in Cape Coast, Ghana are undergoing segregation, which could lead to possible speciation. We also aimed to provide a morphometric description of all nymphal stages. Nine-bed bug populations of C. hemipterus were field-collected in Cape Coast and were subjected to geometric morphometric analysis. The multivariate parameters applied distinguished various populations from each of the locations, indicating the presence of morphologically distinct subpopulations of C. hemipterus. Shape-based segregation and shape changes associated with the insect pronotum (which is an important taxonomic character in the Cimicidae) were evident across the populations. Through this comparative study of C. hemipterus, we showed that possible subpopulations of this bed bug are being spread from Ghana. The nymphal stages (first-fifth) of C. hemipterus were distinguished by the length of the last three antennal segment and pronota width; such information contributes to the taxonomic knowledge of the species.

Geographical patterns and mechanisms of Cimex lectularius Linnaeus, 1758, and Cimex hemipterus Fabricius, 1803 (Hemiptera: Cimicidae) resistance to insecticides: a systematic review and meta-analysis.

This study aimed to review published scientific literature on bed bugs in countries where insecticide resistance has been reported worldwide from 2000 to 2021. Electronic databases, including Scopus, PubMed, and WOS, were searched. Out of 606 articles found in the initial search, we selected 57 articles, of which 40 articles had reported on Cimex lectularius (C. lectularius), and 22 papers had reported on Cimex hemipterus (C. hemipterus). Most studies on insecticide resistance were carried out on C. lectularius in North America (14, 35%) and C. hemipterus in Asia (16, 72.7%). The most common method used to detect bed bug resistance to insecticides was toxicological bioassay with an overall random pooled effect size of 0.38 (95% CI: 0.23-0.53) in C. lectularius and 0.46 (95% CI: 0.27-0.65) in C. hemipterus. Resistance to pyrethroids was reported against C. lectularius with an overall pooled effect size of 0.75 (95% CI: 0.56-0.94) and C. hemipterus with an overall pooled effect size of 0.81 (95% CI: 0.57-0.93) in 33.40 (82.5%) and 19.22 (86.3%) published articles, respectively. A very high resistance level to pyrethroids in both studied species was observed, and resistance ratios at the highest level were 76389.3 and 315.5 in C. lectularius and C. hemipterus, respectively. Resistance mechanisms against pyrethroids were reported from most locations except Iran and Thailand, but these mechanisms were not studied in other insecticide groups. These reports indicate that chemical control options for bed bugs are limited. Therefore, a combination of chemical and non-chemical strategies is recommended for bed bug control.

CT-guided palladium-103 seed brachytherapy for metastatic adenoid cystic carcinoma: a retrospective study to assess initial safety and effectiveness of percutaneous CT fluoroscopy-guided permanent seed brachytherapy.

PURPOSE: Multiple pulmonary metastases present treatment difficulties in available treatment techniques, which are inconvenient or may damage sufficient pulmonary tissue to cause pulmonary crippling. This retrospective study of a single-community practice evaluated responses to computed tomography (CT)-guided 103Pd permanent seed brachytherapy (CTGPSB) in adenoid cystic carcinoma (ACC) synchronous pulmonary metastases. The purpose of the current study was to document that metastatic pulmonary ACC lesions can be controlled with CTGPSB. MATERIAL AND METHODS: Twenty-nine discrete lesions in 14 patients were evaluated with serial CT scans. All were treated with CTGPSB. Lesions were tracked over serial CT scans and volumes measured. Primary endpoint was a reduction in tumor volume on subsequent CT scan. Secondary endpoint was occurrence of CTCAE grades 2-5. RESULTS: There was a 100% measured reduction in tumor volume (n = 29) at follow-up. Follow-up was a mean of 3.13 years. Baseline tumor volume was a mean of 1.85 ml (range, 0.69-9.15 ml). There were two grade 1 and one grade 2 adverse events, which did not require hospitalization. CONCLUSIONS: CTGPSB for the treatment of multiple ACC pulmonary metastases is effective, with minimal acute complications, as shown in small cohort of subjects of the present study. Further studies evaluating specific dosimetry parameters in this free-hand technique are needed to specify minimal and maximal dose constraints.

Multiple Mechanisms Conferring Broad-Spectrum Insecticide Resistance in the Tropical Bed Bug (Hemiptera: Cimicidae).

The modern resurgence of the common (Cimex lectularius L.) and tropical bed bugs (C. hemipterus [F.]) is thought to be primarily due to insecticide resistance. While there are many reports on insecticide resistance mechanisms in C. lectularius, such information in C. hemipterus is limited. We examined dichloro-diphenyl-trichloroethane (DDT), malathion, deltamethrin, permethrin, lambda-cyhalothrin resistance, and the underlying mechanisms in several C. hemipterus strains (Australia: Queensland [QLD-AU]; Malaysia: Kuala Lumpur [KL-MY], Tanjung Tokong [TT-MY], Christian [CH-MY], and Green Lane [GL-MY]). We used a surface contact method, synergism studies (utilizing piperonyl butoxide [PBO], S,S,S-tributyl phosphorotrithioate [DEF], and diethyl maleate [DEM]), and molecular detection of kdr mutations. Results demonstrated that all C. hemipterus strains possessed high resistance to DDT and the pyrethroids and moderate to high resistance to malathion. Synergism studies showed that deltamethrin resistance in all strains was significantly (P < 0.05) inhibited by PBO. In contrast, deltamethrin resistance was not affected in DEF or DEM. Similar findings were found with lambda-cyhalothrin resistance. Malathion resistance was significantly (P < 0.05) reduced by DEF in all strains. Resistance to DDT was not affected by DEM in all strains. Multiple kdr mutations (M918I, D953G, and L1014F) were detected by molecular analyses. TT-MY strain was found with individuals possessing three kdr mutation combinations; D953G + L1014F (homozygous susceptible: M918), M918I + D953G + L1014F (heterozygous resistant: I918), and M918I + D953G + L1014F (homozygous resistant: I918). Individuals with M918I + D953G + L1014F (homozygous resistant: I918) survived longer on deltamethrin (>12 h) than those (≤1 h) with other combinations. M918I + L1014F mutations most likely conferred super-kdr characteristic toward pyrethroids and DDT in C. hemipterus.

The Efficacy of Ultrasonic Pest Repellent Devices against the Australian Paralysis Tick, Ixodes holocyclus (Acari: Ixodidae).

Ultrasonic pest repellers are often promoted as a means of protecting people and pets from the bites of hematophagous arthropods, such as ticks. However, to date, there has been no published research on the effectiveness of these devices against the Australian paralysis tick, Ixodes holocyclus Neumann. The purpose of this study was to test the effectiveness of nine ultrasonic devices against female I. holocyclus. Two arenas were constructed, one for the test (with the ultrasonic device) and one for the control (no device). Each arena had a test and an escape chamber, connected by a corridor. Twenty ticks were placed in each test chamber. After the ultrasonic device was operated for 1 h, the number of ticks in both chambers was recorded. Ten replicates were conducted for each device. The average number of ticks that moved from the test to the escape chamber was greater in all the test arenas, with three devices being statistically different from the control. However, the highest percent of ticks that escaped was only 19.5%. This amount is insufficient to offer adequate protection against tick bites and this study adds further weight to previous investigations that ultrasonic devices should not be employed in pest management.

Assessment of Tropical Bed Bug (Hemiptera: Cimicidae), Infestations in Cape Coast, Ghana: Household Control Practices and Efficacy of Commercial Insecticides and Long-Lasting Insecticidal Nets Against Field Bed Bugs.

This study reports the first baseline information on tropical bed bug, Cimex hemipterus (F.) (Hemiptera: Cimicidae), infestations in Ghana. The purpose of this study was to assess bed bug infestation levels, and the efficacy of locally available insecticides and long-lasting insecticidal nets (LLINs) in controlling field bed bugs populations in the Cape Coast region. A survey was undertaken to assess bed bug infestation levels and current control practices by residents. In total, 205 bed bug affected households were identified in 20 communities and live bed bug infestations were associated with most of these premises. Many homeowners knew of other households (from 1 to 3) with a bed bug infestation. Residents reported itching and swelling of the skin from the bed bug bites and the bites were considered severe. The most common household bed bug control strategy was the application of insecticides. However, LLINs and commercially formulated insecticides commonly used by households (notably chloropyrifos and pyrethroid-based formulations) did not efficaciously suppress field collected strains of C. hemipterus. Using a dipping bioassay, mean mortality ranged from 0 to 60% for eggs, nymphs, and adults, and less than 40% mortality was observed in bed bugs placed on insecticide-treated filter paper. Each LLINs (all are pyrethroid based) produced a mean mortality of less than 20% in adult bed bugs. For a more effective response to the global bed bug resurgence in developing countries, government and supporting agencies need to render assistance to bed bug affected residents through the provision of improved pest management strategies.

Genetic, Morphological and Antigenic Relationships between Mesonivirus Isolates from Australian Mosquitoes and Evidence for Their Horizontal Transmission.

The Mesoniviridae are a newly assigned family of viruses in the order Nidovirales. Unlike other nidoviruses, which include the Coronaviridae, mesoniviruses are restricted to mosquito hosts and do not infect vertebrate cells. To date there is little information on the morphological and antigenic characteristics of this new group of viruses and a dearth of mesonivirus-specific research tools. In this study we determined the genetic relationships of recent Australian isolates of Alphamesonivirus 4 (Casuarina virus-CASV) and Alphamesonivirus 1 (Nam Dinh virus-NDiV), obtained from multiple mosquito species. Australian isolates of NDiV showed high-level similarity to the prototype NDiV isolate from Vietnam (99% nucleotide (nt) and amino acid (aa) identity). Isolates of CASV from Central Queensland were genetically very similar to the prototype virus from Darwin (95-96% nt and 91-92% aa identity). Electron microscopy studies demonstrated that virion diameter (≈80 nm) and spike length (≈10 nm) were similar for both viruses. Monoclonal antibodies specific to CASV and NDiV revealed a close antigenic relationship between the two viruses with 13/34 mAbs recognising both viruses. We also detected NDiV RNA on honey-soaked nucleic acid preservation cards fed on by wild mosquitoes supporting a possible mechanism of horizontal transmission between insects in nature.

A Unique Relative of Rotifer Birnavirus Isolated from Australian Mosquitoes.

The family Birnaviridae are a group of non-enveloped double-stranded RNA viruses which infect poultry, aquatic animals and insects. This family includes agriculturally important pathogens of poultry and fish. Recently, next-generation sequencing technologies have identified closely related birnaviruses in Culex, Aedes and Anopheles mosquitoes. Using a broad-spectrum system based on detection of long double-stranded RNA, we have discovered and isolated a birnavirus from Aedes notoscriptus mosquitoes collected in northern New South Wales, Australia. Phylogenetic analysis of Aedes birnavirus (ABV) showed that it is related to Rotifer birnavirus, a pathogen of microscopic aquatic animals. In vitro cell infection assays revealed that while ABV can replicate in Aedes-derived cell lines, the virus does not replicate in vertebrate cells and displays only limited replication in Culex- and Anopheles-derived cells. A combination of SDS-PAGE and mass spectrometry analysis suggested that the ABV capsid precursor protein (pVP2) is larger than that of other birnaviruses and is partially resistant to trypsin digestion. Reactivity patterns of ABV-specific polyclonal and monoclonal antibodies indicate that the neutralizing epitopes of ABV are SDS sensitive. Our characterization shows that ABV displays a number of properties making it a unique member of the Birnaviridae and represents the first birnavirus to be isolated from Australian mosquitoes.

Influences of Exposure Time and Mortality Assessment Interval on Bioassay Results of Insecticide-Resistant Tropical Bed Bugs (Hemiptera: Cimicidae).

We evaluated the influences of insecticide exposure time and mortality assessment intervals on the bioassay results of three insecticide-resistant tropical bed bug Cimex hemipterus (F.) populations (Madam Mo, Tanjong Tokong, and Green Lane). This was achieved using the surface contact method and tested with six commercial insecticide products: Tandem, Temprid SC, Pesguard FG161, Sumithrin, Sumithion, and Phantom applied at label rate on glass petri dishes. Six exposure times (5 min, 10 min, 30 min, 1 h, 4 h, and continuous exposure for 4 or 14 days for Phantom) were tested. A susceptible common bed bug Cimex lectularius L. strain (Monheim) was used as the reference strain as no susceptible C. hemipterus strain exists. In treatment with Temprid SC, the Tanjong Tokong strain showed significantly higher KT50 values at 5, 10, and 30 min exposures than 1 h, 4 h, and continuous exposures. When all resistant populations were evaluated at continuous exposure to Sumithion, they demonstrated moderate resistance levels (RR50 ranged from 6.0 to 7.9), while KT50 and KT95 of other shorter exposure times failed to be generated due to low knockdown rate. Higher mortalities were observed in Tanjong Tokong and Green Lane strains when tested at longer exposure times with Temprid SC, Pesguard FG161, Sumithrin, Sumithion, and Phantom. Better killing effect was observed in the treatment with Temprid SC (Tanjong Tokong and Green Lane strains), Pesguard FG161 (Tanjong Tokong and Green Lane strains), Sumithrin (all C. hemipterus strains), Sumithion (all C. hemipterus strains), and Phantom (all strains tested) at longer mortality assessment intervals. We demonstrated that insecticide exposure time and mortality assessment interval could potentially affect outcomes of product performance evaluations, resulting in underestimation or overestimation of insecticide resistance levels in field populations.

Identification of Natural Molecular Determinants of Ross River Virus Type I Interferon Modulation.

Ross River virus (RRV) belongs to the genus Alphavirus and is prevalent in Australia. RRV infection can cause arthritic symptoms in patients and may include rash, fever, arthralgia, and myalgia. Type I interferons (IFN) are the primary antiviral cytokines and trigger activation of the host innate immune system to suppress the replication of invading viruses. Alphaviruses are able to subvert the type I IFN system, but the mechanisms used are ill defined. In this study, seven RRV field strains were analyzed for induction of and sensitivity to type I IFN. The sensitivities of these strains to human IFN-ß varied significantly and were highest for the RRV 2548 strain. Compared to prototype laboratory strain RRV-T48, RRV 2548 also induced higher type I IFN levels both in vitro and in vivo and caused milder disease. To identify the determinants involved in type I IFN modulation, the region encoding the nonstructural proteins (nsPs) of RRV 2548 was sequenced, and 42 amino acid differences from RRV-T48 were identified. Using fragment swapping and site-directed mutagenesis, we discovered that substitutions E402A and R522Q in nsP1 as well as Q619R in nsP2 were responsible for increased sensitivity of RRV 2548 to type I IFN. In contrast, substitutions A31T, N219T, S580L, and Q619R in nsP2 led to induction of higher levels of type I IFN. With exception of E402A, all these variations are common for naturally occurring RRV strains. However, they are different from all known determinants of type I IFN modulation reported previously in nsPs of alphaviruses.IMPORTANCE By identifying natural Ross River virus (RRV) amino acid determinants for type I interferon (IFN) modulation, this study gives further insight into the mechanism of type I IFN modulation by alphaviruses. Here, the crucial role of type I IFN in the early stages of RRV disease pathogenesis is further demonstrated. This study also provides a comparison of the roles of different parts of the RRV nonstructural region in type I IFN modulation, highlighting the importance of nonstructural protein 1 (nsP1) and nsP2 in this process. Three substitutions in nsP1 and nsP2 were found to be independently associated with enhanced type I IFN sensitivity, and four independent substitutions in nsP2 were important in elevated type I IFN induction. Such evidence has clear implications for RRV immunobiology, persistence, and pathology. The identification of viral proteins that modulate type I IFN may also have importance for the pathogenesis of other alphaviruses.

Performance of Commercial Insecticide Formulations Against Different Developmental Stages of Insecticide-Resistant Tropical Bed Bugs (Hemiptera: Cimicidae).

This study examined the presence of insecticide resistance in different developmental stages (adults, first instars, and eggs) of the tropical bed bug, Cimex hemipterus (F.) using several insecticide formulations. Adults and first instars of five strains (Queensland, Kuala Lumpur, Bukit Mertajam, Saujana, and Krystal Point) were evaluated using the surface contact method and compared with a susceptible strain (Monheim) of the common bed bug Cimex lectularius L. The insecticide formulations were used at their label rates in this study: Tandem (thiamethoxam [11.6%], lambda-cyhalothrin [3.5%]) at 183.96 mg/m2; Temprid SC (imidacloprid [21%], beta-cyfluthrin [10.5%]) at 106.13 mg/m2; Sumithion 20CS (fenitrothion [20%]) at 250 mg/m2; Pesguard FG161 (d-tetramethrin [4.4%], cyphenothrin [13.2%]) at 110 mg/m2; and Sumithrin 10SEC (d-phenothrin [10%]) at 100 mg/m2. Results showed a very high level of resistance to Pesguard FG161 (388.3 to >605.0 times) and Sumithrin (302.9 to >365.5 times) in all adults of the strains tested, whereas low to high levels of resistance were registered for Tandem (1.4-4.7 times), Temprid (7.3-16.7 times), and Sumithion (1.2-14.6 times) for adults of all bed bug strains. For first instars, resistance to the former two formulations were high to very high (31.4-118.1 times). In contrast, they showed lower resistance to Tandem, Temprid, and Sumithion (1.0-10.2 times). An immersion method used to test on bed bug eggs found high to very high resistance toward all tested formulations. Results demonstrate that the resistance level varies between bed bug developmental stages.