Peptide toxins that target vertebrate voltage-gated sodium channels underly the painful stings of harvester ants.

Harvester ants (genus Pogonomyrmex) are renowned for their stings which cause intense, long-lasting pain, and other neurotoxic symptoms in vertebrates. Here, we show that harvester ant venoms are relatively simple and composed largely of peptide toxins. One class of peptides is primarily responsible for the long-lasting local pain of envenomation via activation of peripheral sensory neurons. These hydrophobic, cysteine-free peptides potently modulate mammalian voltage-gated sodium (NaV) channels, reducing the voltage threshold for activation and inhibiting channel inactivation. These toxins appear to have evolved specifically to deter vertebrates.

Venom chemistry underlying the painful stings of velvet ants (Hymenoptera: Mutillidae).

Velvet ants (Hymenoptera: Mutillidae) are a family of solitary parasitoid wasps that are renowned for their painful stings. We explored the chemistry underlying the stings of mutillid wasps of the genus Dasymutilla Ashmead. Detailed analyses of the venom composition of five species revealed that they are composed primarily of peptides. We found that two kinds of mutillid venom peptide appear to be primarily responsible for the painful effects of envenomation. These same peptides also have defensive utility against invertebrates, since they were able to incapacitate and kill honeybees. Both act directly on cell membranes where they directly increase ion conductivity. The defensive venom peptides of Dasymutilla bear a striking similarity, in structure and mode of action, to those of the ant Myrmecia gulosa (Fabricius), suggesting either retention of ancestral toxins, or convergence driven by similar life histories and defensive selection pressures. Finally, we propose that other highly expressed Dasymutilla venom peptides may play a role in parasitisation, possible in delay or arrest of host development. This study represents the first detailed account of the composition and function of the venoms of the Mutillidae.

Rapid detection of human blood in triatomines (kissing bugs) utilizing a lateral flow immunochromatographic assay - A pilot study.

OBJECTIVES: We tested a rapid and specific immunochromatographic assay (that detects human blood in forensic samples) to determine if human blood was present in triatomines and their fecal excreta. METHODS: We fed Triatoma rubida human blood (positive control) or mouse blood (negative control) and performed the assay on the abdominal contents and fecal excreta. Triatomine field specimens collected in and around human habitations and excreta were also tested. FINDINGS: The assay was positive in triatomines fed human blood (N = 5/5) and fecal excreta from bugs known to have ingested human blood (N = 5/5). Bugs feeding on mice (N = 15/15) and their fecal excreta (N = 8/8) were negative for human blood. Human blood was detected in 47% (N = 23/49) triatomines, representing six different species, collected in the field. MAIN CONCLUSIONS: The pilot study shows that this rapid and specific test may have applications in triatomine research. Further study is needed to determine the sensitivity of this assay compared to other well-established techniques, such as DNA- and proteomics-based methodologies and the assay's application in the field.

Protein mass spectrometry extends temporal blood meal detection over polymerase chain reaction in mouse-fed Chagas disease vectors.

BACKGROUND: Chagas disease is highly prevalent in Latin America, and vector control is the most effective control strategy to date. We have previously shown that liquid chromatography tandem mass spectrometry (LC-MS/MS) is a valuable tool for identifying triatomine vector blood meals. OBJECTIVES: The purpose of this study was to determine blood meal detection ability as a function of method [polymerase chain reaction (PCR) vs. LC-MS/MS], time since feeding, and the effect of molting in mouse-fed triatomine insect vectors targeting hemoglobin and albumin proteins with LC-MS/MS and short interspersed nuclear elements (SINE)-based PCR. METHODS: We experimentally fed Triatoma protracta on mice and used LC-MS/MS to detect hemoglobin and albumin peptides over time post-feeding and post-molting (≤ 12 weeks). We compared LC-MS/MS results with those of a standard PCR method based on SINEs. FINDINGS: Hemoglobin-based LC-MS/MS detected blood meals most robustly at all time points post-feeding. Post-molting, no blood meals were detected with PCR, whereas LC-MS/MS detected mouse hemoglobin and albumin up to 12 weeks. MAIN CONCLUSIONS: In our study, the hemoglobin signature in the insect abdomen lasted longer than that of albumin and DNA. LC-MS/MS using hemoglobin shows promise for identifying triatomine blood meals over long temporal scales and even post-molting. Clarifying the frequency of blood-feeding on different hosts can foster our understanding of vector behavior and may help devise sounder disease-control strategies, including Ecohealth (community based ecosystem management) approaches.

Evolutionary responses of solitary and social Hymenoptera to predation by primates and overwhelmingly powerful vertebrate predators.

Insects provide an important part of the diet of primates, including hominins. Investigations of insectivory in primates has focused primarily on the value of insects in the diet, and on the means of obtaining the insects, with little attention devoted to the predator-prey relationship itself and less to evolutionary aspects of insect defense against predatory vertebrates, including primates. Data indicate that, far from being a passive half of the relationship, insects in general, and stinging Hymenoptera in particular, are active participants that have greatly influenced the relationship. Predators have been a strong component of the selection pressure in the evolution of painful and toxic bee, wasp, and ant stings and these insects, in turn, have influenced hunting behavior and learning in at least higher primates. The special example of honey bees and humans is highlighted. Both humans and the bees have benefitted from a relationship that represents an unprecedented example of a predator-prey interaction evolving recently into facultative mutualism.

Veteran influenza vaccination acceptance rates after completion of the COVID-19 vaccination series among historical influenza vaccine refusers.

BACKGROUND: Vaccine hesitancy remains an obstacle in disease prevention. The recent COVID-19 pandemic highlighted this issue and may influence acceptance of other recommended immunizations. The objective of this study was to determine the association between receiving the COVID-19 vaccination and the subsequent acceptance of the influenza vaccination in a Veteran population that historically declined influenza vaccination. METHODS: Influenza vaccination acceptance rates for the 2021-2022 influenza season were compared in patients who historically declined the influenza vaccine and either received or declined COVID-19 vaccinations. Logistic regression analysis was used to analyze factors associated with receiving influenza vaccination among vaccine hesitant individuals. RESULTS: A higher proportion of patients who had received the COVID-19 vaccination(s) subsequently accepted the influenza vaccination compared to the control group (37% vs. 11%, OR = 5.03; CI 3.15-8.26; p = 0.0001). CONCLUSION: Among previous influenza vaccine decliners, those who received COVID-19 vaccination had significantly higher odds of receiving subsequent influenza vaccination.

Functional and Proteomic Insights into Aculeata Venoms.

Aculeate hymenopterans use their venom for a variety of different purposes. The venom of solitary aculeates paralyze and preserve prey without killing it, whereas social aculeates utilize their venom in defence of their colony. These distinct applications of venom suggest that its components and their functions are also likely to differ. This study investigates a range of solitary and social species across Aculeata. We combined electrophoretic, mass spectrometric, and transcriptomic techniques to characterize the compositions of venoms from an incredibly diverse taxon. In addition, in vitro assays shed light on their biological activities. Although there were many common components identified in the venoms of species with different social behavior, there were also significant variations in the presence and activity of enzymes such as phospholipase A2s and serine proteases and the cytotoxicity of the venoms. Social aculeate venom showed higher presence of peptides that cause damage and pain in victims. The venom-gland transcriptome from the European honeybee (Apis mellifera) contained highly conserved toxins which match those identified by previous investigations. In contrast, venoms from less-studied taxa returned limited results from our proteomic databases, suggesting that they contain unique toxins.

Ant venoms contain vertebrate-selective pain-causing sodium channel toxins.

Stings of certain ant species (Hymenoptera: Formicidae) can cause intense, long-lasting nociception. Here we show that the major contributors to these symptoms are venom peptides that modulate the activity of voltage-gated sodium (NaV) channels, reducing their voltage threshold for activation and inhibiting channel inactivation. These peptide toxins are likely vertebrate-selective, consistent with a primarily defensive function. They emerged early in the Formicidae lineage and may have been a pivotal factor in the expansion of ants.

Vector blood meals and Chagas disease transmission potential, United States.

A high proportion of triatomine insects, vectors for Trypanosoma cruzi trypanosomes, collected in Arizona and California and examined using a novel assay had fed on humans. Other triatomine insects were positive for T. cruzi parasite infection, which indicates that the potential exists for vector transmission of Chagas disease in the United States.

Seasonal Flight Pattern of the Kissing Bugs Triatoma rubida and T. protracta (Hemiptera: Reduviidae: Triatominae) in Southern Arizona, United States.

The two most common kissing bugs, Triatoma rubida and T. protracta, in the Sonoran Desert around Tucson, Arizona are hematophagous vectors of Chagas disease and can induce potentially life-threatening allergic reactions. They were surveyed during their summer dispersal flight period to determine which environmental factors are correlated with flight activity. The two most important factors governing flights of T.rubida were temperatures in the range of 26-35 °C and wind speeds below 14 km/h (9 miles/h). Flights were reduced below or above those temperatures, or when wind speeds exceeding 14km/h. Relative humidity and presence or absence of moonshine appeared unimportant. During their dispersal flight periods of May through July and, especially, between the peak of the flight season, 20 June to 5 July, biologists seeking to collect bugs and homeowners wishing to exclude these biting bugs from entering their homes should be most attentive during evenings of average temperature and low wind speed.

Kissing Bug Intrusions into Homes in the Southwest United States.

Kissing bugs readily enter homes in the Sonoran Desert and bite the residents. Their saliva is highly antigenic, causing local and systemic skin reactions and life-threatening anaphylaxis. We attempted to determine what characteristics of homesites may have contributed to home intrusion by kissing bugs. Extensive and detailed information about the homes and the home environment was collected from 78 homeowners in Tucson who suffered kissing bug intrusions. Homeowners collected 298 Triatoma rubida in and around their homes. Of the homes entered by kissing bugs, 29 of 46 (63%) contained bugs harboring Trypanosoma cruzi. Although in the aggregate, homeowners were bitten > 2200 times, no individual tested positive for Chagas disease (N = 116). Although yearly intrusion likely occurs in some homes, T. rubida does not domiciliate within homesites in the Desert Southwest. We conclude there is little risk to homeowners for Chagas disease given the current behavior of resident kissing bugs and absent ingesting kissing bug fecal matter.

Scorpion Stings and Antivenom Use in Arizona.

BACKGROUND: Arizona's rugged desert landscape harbors many venomous animals, including a small nocturnal scorpion, Centruroides sculpturatus, whose venom can cause severe neuromotor disturbance. An effective antivenom is available at selected health care facilities in the state. METHODS: We analyzed 4398 calls of scorpion stings to the Arizona Poison and Drug Information Center (APDIC) in Tucson over a period of 3 years, from January 2017 to December 2019. RESULTS: We followed 1952 (44.4%) of the victims to resolution. We excluded 2253 callers with minimal effects of the sting and 193 victims with possible toxic effects who were lost to follow-up. The most common complaints among callers were pain at the sting site in 88.9% and local numbness in 62.2%. Detailed clinical information was obtained from 593 calls from a health care facility. Neuromotor signs consistent with C. sculpuratus envenomation included nystagmus in 163 (27.5%), hypersalivation in 91 (15.3%), and fasciculations in 88 (14.8%). Antivenom (Anascorp; Rare Disease Therapeutics, Inc., Franklin, Tenn) was administered to 145 patients. Most were children <5 years old (n = 76, or 54.4%); 27 (18.6%) were 5-9 years old and 42 (30.0%) were ≥10 years of age. About half, 79 of 145 (54.5%) victims who received antivenom, met the APDIC recommended use criteria. CONCLUSIONS: Patients treated with antivenom exhibited a rapid resolution of symptoms without immediate or delayed hypersensitivity reactions. We recommend broadened availability of antivenom at sites where it is most needed.

A Southwestern United States Pilot Investigation of Triatomine-Mite Prevalence.

BACKGROUND: Chagas disease is a leading cause of cardiac failure in Latin America. Due to poor safety profiles and efficacy of currently available therapeutics, prevention is a priority for the millions living at risk for acquiring this clinically important vector-borne disease. Triatomine vectors of the Chagas disease parasite, Trypanosoma cruzi, are found in the southwestern United States, but risk for autochthonous transmission is thought to be low. The role of ectoparasitic mites is under-explored regarding the ecology of triatomines and Chagas disease transmission. METHODS: Triatomine collections were performed using three common entomologic techniques in 2020-2021 from four different locations in southern Arizona and New Mexico. Triatomines were analyzed visually under a 112.5× microscope for the presence of externally attached mites. Following mite removal, triatomines were tested for T. cruzi infection by PCR. RESULTS: Approximately 13% of the collected triatomines had mites securely attached to their head, thorax, abdomen, and legs. More than one mite attached was a common finding among ectoparasitized triatomines. Mite presence, however, did not statistically influence triatomine T. cruzi status. CONCLUSIONS: Our findings add to a growing body of literature demonstrating the sustainability of mite-infested triatomine populations throughout the Western Hemisphere. Future investigations are warranted to better understand the biologic impact of triatomine mites and their potential to serve as a potential biological control tool.

"Kissing bugs": potential disease vectors and cause of anaphylaxis.

Physicians in the United States should familiarize themselves with "kissing bugs" endemic to their area of practice and appreciate the medical implications of their bites. Bite victims often seek advice from physicians about allergic reactions as well as the risk of contracting Chagas disease. Physicians are generally knowledgeable about the role of kissing bugs in the transmission of Trypanosoma cruzi in Latin America. However, they may be unaware of (1) severe allergic reactions to kissing bug salivary antigens, (2) the widespread occurrence of T. cruzi amongst vertebrate hosts of kissing bugs, and (3) the incidence of T. cruzi among kissing bugs (T. cruzi may infect >50% of sampled bugs). Despite the potential for Chagas disease transmission, the major concern regarding kissing bugs in the United States is anaphylactic reactions to their bites resulting in frequent emergency department visits, especially in areas of endemicity in the Southwest.

Autochthonous Chagas Disease: How Are These Infections Happening?

Proven cases of vector-transmitted acute autochthonous Chagas disease in the United States are rare (<10 cases). Possible or probable cases of unknown duration determined by serology are uncommon as well (<100). In Latin America it is widely accepted that after feeding, the kissing bug defecates and Trypanosoma cruzi in the feces is rubbed into the bite punctum. This is an inefficient method of parasite transmission. The average infected individual in Latin America suffers more than 1000 bites, but more importantly, there are often thousands of kissing bugs in a household dropping feces on the inhabitants and living quarters. Today in Brazil the most common form of acute Chagas is secondary to oral ingestion of the parasite in food and drink. We present our experience with many hundreds of individuals bitten by kissing bugs and the possibility of oral ingestion occurring in the United States.

Honeybee Stings in the Era of Killer Bees: Anaphylaxis and Toxic Envenomation.

BACKGROUND: Twenty-six years after the arrival of "killer bees" in Arizona, the entire state with the exception of high elevations in the north is populated with this bee variety and 11 people have died at the scene of massive bee attacks. METHODS: Because of the aggressive behavior of these bees we studied bee stings reported to the Arizona Poison and Drug Information Center. The center received 399 calls regarding 312 victims of bee stings from January 2017 to June 2019. Calls originated from private residences and emergency centers. RESULTS: Stings occurred at victims' home residences in 272 (84.7%) of cases and 24 (7.5%) in public areas; 251 people suffered 1 sting; 42 individuals, 2-10 stings, 4 had 11-49 stings, and 13 individuals had >50 stings (so-called massive stinging). Three individuals were admitted to intensive care units (ICU) and one 35-year-old man died of anaphylaxis after 1 sting; moderate clinical effects occurred in 32 individuals including 6 admitted to the hospital but not in the intensive care unit. Anaphylaxis occurred in 30 (9.6%) of individuals, 16 receiving 1 sting. Toxic effects, tachycardia, elevated creatinine, or rhabdomyolysis occurred in 13 (4.2%) individuals. CONCLUSIONS: In the past, individuals stung more than 50 times were beekeepers working with European honeybees, whereas, in the current era, single as well as massive stings are the result of feral "killer bees." This change in epidemiology requires a new approach to sting victims: those with massive stinging should be evaluated and observed for anaphylaxis and serial laboratory values obtained for days to detect the toxic effects of envenomation.

Kissing Bugs Harboring Trypanosoma cruzi, Frequently Bite Residents of the US Southwest But Do Not Cause Chagas Disease.

BACKGROUND: Kissing bugs are common household pests in the Desert Southwest of the United States. These hematophagous bugs enter homes and suck blood from resident humans and pets. They are vectors of Trypanosoma cruzi, an enzootic parasite in small mammals and the cause of Chagas disease in humans. Autochthonous cases of Chagas disease are rare in the United States despite the presence of the vector and parasite. Environmental and biological factors accounting for this phenomenon need studying. METHODS: Homeowners in Bisbee and Tucson, Arizona captured kissing bugs inside homes during 2017-2018. Bugs were tested for presence of T. cruzi by polymerase chain reaction. Residents bitten by kissing bugs were tested for Chagas disease by serology. We evaluated invaded homes in the 2 cities. RESULTS: Three species of kissing bugs (n = 521) were collected in or near homes. Triatoma rubida was the most common triatomine in Tucson; T. recurva in Bisbee. T. protracta was uncommon. Seventeen percent of bugs captured in Bisbee and 51.1% in Tucson harbored T. cruzi. Bite victims (n = 105) recalled more than 2200 bites. Reactions to bites were common, including 32 episodes of anaphylaxis in 11 people (10.5%). Tests for Chagas disease (n = 116) were negative. Median age of houses was 91 years in Bisbee and 7 years in Tucson. Bisbee houses had pier and beam foundations. Tucson houses were built on concrete slabs. CONCLUSIONS: Kissing bugs harboring T. cruzi readily entered new and old homes. Bites of humans caused severe, life-threatening reactions. There was no serological evidence of Chagas disease among those bitten.

Barriers to and factors influencing the pursuit of pharmacy student research.

INTRODUCTION: The objective of this study was to identify perceived barriers and factors influencing student pursuit of research during pharmacy school. METHODS: A voluntary, paper-based or electronic questionnaire was administered to all pharmacy students at a private college of pharmacy in mandatory courses during the 2016-2017 academic year. Survey questions collected information pertaining to demographics, factors influencing student pursuit of research, and barriers to pursuit of research. Participation was incentivized with gift cards. RESULTS: A total of 623 students completed the survey (79% response rate). The average respondent was female (69.1%), 25-years old (IQR 23-26 years), employed (69.9%), and had a prior degree (66.6%). During pharmacy school, 27.3% of respondents pursued a research project. Of students not pursuing a research project during pharmacy school, the amount of interest to complete a project differed significantly between professional years (P < .01) with the second-year class having the highest interest. Lack of time (91.3%), unfamiliarity with the research process (81.8%), and too much coursework (80.5%) were cited as the top three perceived barriers that prevented students from pursuing research. A mandatory research class (87.6%), presentations describing faculty research interests (83.4%), and ability to work with a friend (83.9%) would most strongly influence students to complete research. CONCLUSIONS: Students report additional information regarding research opportunities would positively influence their decision to pursue research while in pharmacy school. Future studies should evaluate strategies to familiarize students with the research process.