Pediatric and adult patients with ME/CFS following COVID-19: A structured approach to diagnosis using the Munich Berlin Symptom Questionnaire (MBSQ).

A subset of patients with post-COVID-19 condition (PCC) fulfill the clinical criteria of myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS). To establish the diagnosis of ME/CFS for clinical and research purposes, comprehensive scores have to be evaluated. We developed the Munich Berlin Symptom Questionnaires (MBSQs) and supplementary scoring sheets (SSSs) to allow for a rapid evaluation of common ME/CFS case definitions. The MBSQs were applied to young patients with chronic fatigue and post-exertional malaise (PEM) who presented to the MRI Chronic Fatigue Center for Young People (MCFC). Trials were retrospectively registered (NCT05778006, NCT05638724). Using the MBSQs and SSSs, we report on ten patients aged 11 to 25 years diagnosed with ME/CFS after asymptomatic SARS-CoV-2 infection or mild to moderate COVID-19. Results from their MBSQs and from well-established patient-reported outcome measures indicated severe impairments of daily activities and health-related quality of life.    Conclusions: ME/CFS can follow SARS-CoV-2 infection in patients younger than 18 years, rendering structured diagnostic approaches most relevant for pediatric PCC clinics. The MBSQs and SSSs represent novel diagnostic tools that can facilitate the diagnosis of ME/CFS in children, adolescents, and adults with PCC and other post-infection or post-vaccination syndromes. What is Known: • ME/CFS is a debilitating disease with increasing prevalence due to COVID-19. For diagnosis, a differential diagnostic workup is required, including the evaluation of clinical ME/CFS criteria. • ME/CFS after COVID-19 has been reported in adults but not in pediatric patients younger than 19 years. What is New: • We present the novel Munich Berlin Symptom Questionnaires (MBSQs) as diagnostic tools to assess common ME/CFS case definitions in pediatric and adult patients with post-COVID-19 condition and beyond. • Using the MBSQs, we diagnosed ten patients aged 11 to 25 years with ME/CFS after asymptomatic SARS-CoV-2 infection or mild to moderate COVID-19.

Common double-lumen tube selection methods overestimate adequate tube sizes in individual patients - a 3D reconstruction study.

BACKGROUND: Appropriate selection of double-lumen tube sizes for one-lung ventilation is crucial to prevent airway damage. Current selection methods rely on demographic factors or 2D radiography. Prediction of left bronchial diameter is indispensable for choosing the adequate tube size. This prospective observational study investigates if current selection methods sufficiently predict individuals' left bronchial diameters for DLT selection compared to the 3D reconstruction. METHODS: 100 patients necessitating thoracic surgery with one-lung ventilation and left-sided double-lumen tubes, ≥ 18 years of age, and a set of chest X-rays and 2D thorax CT scans for 3D reconstruction of the left main bronchus were included between 07/2021 and 06/2023. The cross-validated prediction error and the width of the 95%-prediction intervals of the 3D left main bronchial diameter utilizing linear prediction models were based on current selection methods. RESULTS: The mean bronchial diameter in 3D reconstruction was 13.6 ± 2.1 mm. The ranges of the 95%-prediction intervals for the bronchial diameter were 6.4 mm for demographic variables, 8.3 mm for the tracheal diameter from the X-ray, and 5.9 mm for bronchial diameter from the 2D-CT scans. Current methods violated the suggested '≥1 mm' safety criterion in up to 7% (men) and 42% (women). Particularly, 2D radiography overestimated women's left bronchial diameter. Current methods even allowed the selection of double-lumen tubes with bronchial tube sections greater than the bronchial diameter in women. CONCLUSIONS: Neither demographic nor 2D-radiographic methods sufficiently account for the variability of the bronchial diameter. Wide 95%-prediction intervals for the bronchial diameter hamper accurate individual double-lumen tube selection. This increases women's risk of bronchial damage, particularly if they have other predisposing factors. These patients may benefit from 3D reconstruction of the left main bronchus. TRIAL REGISTRATION: Not applicable.

Paired methods to measure biofilm killing and removal: a case study with Penicillin G treatment of Staphylococcus aureus biofilm.

Biofilms are microbial aggregates that show high tolerance to antibiotic treatments in vitro and in vivo. Killing and removal are both important in biofilm control, therefore methods that measure these two mechanisms were evaluated in a parallel experimental design. Kill was measured using the single tube method (ASTM method E2871) and removal was determined by video microscopy and image analysis using a new treatment flow cell. The advantage of the parallel test design is that both methods used biofilm covered coupons harvested from a CDC biofilm reactor, a well-established and standardized biofilm growth method. The control Staphylococcus aureus biofilms treated with growth medium increased by 0·6 logs during a 3-h contact time. Efficacy testing showed biofilms exposed to 400 µmol l-1 penicillin G decreased by only 0·3 logs. Interestingly, time-lapse confocal scanning laser microscopy revealed that penicillin G treatment dispersed the biofilm despite being an ineffective killing agent. In addition, no biofilm removal was detected when assays were performed in 96-well plates. These results illustrate that biofilm behaviour and impact of treatments can vary substantially when assayed by different methods. Measuring both killing and removal with well-characterized methods will be crucial for the discovery of new anti-biofilm strategies. SIGNIFICANCE AND IMPACT OF THE STUDY: Biofilms are tolerant to antimicrobial treatments and can lead to persistent infections. Finding new anti-biofilm strategies and understanding their mode-of-action is therefore of high importance. Historically, antimicrobial testing has focused on measuring the decrease in viability. While kill data are undeniably important, measuring biofilm disruption provides equally useful information. Starting with biofilm grown in the same reactor, we paired assessment of biofilm removal using a new treatment-flow-cell and real-time microscopy with kill data collected using the single tube method (ASTM E2871). Pairing these two methods revealed efficient biofilm removal properties of Penicillin G which were not detected during efficacy testing.

The effect of skin-to-skin care on cerebral oxygenation during nasogastric feeding of preterm infants.

AIM: To describe cerebral oxygenation during gavage feeding of preterm infants during incubator and skin-to-skin care. METHODS: Further analysis of data from two crossover studies comparing cerebral oxygenation, heart rate and oxygen saturation during skin-to-skin care with incubator care. Data were analysed in three epochs; 10 minutes prefeed, during-feed and 10 minutes postfeed. Measurements from infants fed during incubator care were compared with those obtained during skin-to-skin care. RESULTS: In 39 infants [median (IQR) 27.8 (26.1-30.0) weeks' gestation], there was no difference in cerebral oxygenation between pre-, during- and postfeed. Heart rate increased by three beats per minute postfeed compared with during-feed. Twenty infants received two gavage feeds, one feed in the incubator and another during skin-to-skin care. There was no difference in cerebral oxygenation and heart rate; peripheral oxygen saturation decreased by 3% during feeding whilst skin-to-skin care compared with feeding in the incubator. CONCLUSION: Cerebral oxygenation remained stable before, during and after gavage feeding in an incubator and during skin-to-skin care. The small decrease in oxygen saturation whilst receiving gavage feeding during skin-to-skin care is unlikely to be clinically important, providing reassurance that preterm infants maintain physiological stability during skin-to-skin care.

The German Multicenter Registry for ME/CFS (MECFS-R).

Background: Myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS) is a debilitating multisystemic disease characterized by a complex, incompletely understood etiology. Methods: To facilitate future clinical and translational research, a multicenter German ME/CFS registry (MECFS-R) was established to collect comprehensive, longitudinal, clinical, epidemiological, and laboratory data from adults, adolescents, and children in a web-based multilayer-secured database. Results: Here, we present the research protocol and first results of a pilot cohort of 174 ME/CFS patients diagnosed at two specialized tertiary fatigue centers, including 130 (74.7%) adults (mean age 38.4; SD 12.6) and 43 (25.3%) pediatric patients (mean age 15.5; SD 4.2). A viral trigger was identified in 160/174 (92.0%) cases, with SARS-CoV-2 in almost half of them. Patients exhibited severe functional and social impairment, as reflected by a median Bell Score of 30.0 (IQR 30.0 to 40.0) and a poor health-related quality of life assessed with the Short Form-36 health survey, resulting in a mean score of 40.4 (SD 20.6) for physical function and 59.1 (SD 18.8) for mental health. Conclusions: The MECFS-R provides important clinical information on ME/CFS to research and healthcare institutions. Paired with a multicenter biobank, it facilitates research on pathogenesis, diagnostic markers, and treatment options. Trial registration: ClinicalTrials.gov NCT05778006.

Effects of minute misregistrations of prefabricated markers for image-guided dental implant surgery: an analytical evaluation.

OBJECTIVES: The goal of the present study was to develop a theoretical analysis of errors in implant position, which can occur owing to minute registration errors of a reference marker in a cone beam computed tomography volume when inserting an implant with a surgical stent. MATERIAL AND METHODS: A virtual dental-arch model was created using anatomic data derived from the literature. Basic trigonometry was used to compute effects of defined minute registration errors of only voxel size. The errors occurring at the implant's neck and apex both in horizontal as in vertical direction were computed for mean ±95%-confidence intervals of jaw width and length and typical implant lengths (8, 10 and 12 mm). RESULTS: Largest errors occur in vertical direction for larger voxel sizes and for greater arch dimensions. For a 10 mm implant in the frontal region, these can amount to a mean of 0.716 mm (range: 0.201-1.533 mm). Horizontal errors at the neck are negligible, with a mean overall deviation of 0.009 mm (range: 0.001-0.034 mm). Errors increase with distance to the registration marker and voxel size and are affected by implant length. CONCLUSION: Our study shows that minute and realistic errors occurring in the automated registration of a reference object have an impact on the implant's position and angulation. These errors occur in the fundamental initial step in the long planning chain; thus, they are critical and should be made aware to users of these systems.

Investigating the relationship of COVID-19 related stress and media consumption with schizotypy, depression, and anxiety in cross-sectional surveys repeated throughout the pandemic in Germany and the UK.

Background: Studies report a strong impact of the COVID-19 pandemic and related stressors on the mental well-being of the general population. In this paper, we investigated whether COVID-19 related concerns and social adversity affected schizotypal traits, anxiety, and depression using structural equational modelling. In mediation analyses, we furthermore explored whether these associations were mediated by healthy (sleep and physical exercise) or unhealthy behaviours (drug and alcohol consumption, excessive media use). Methods: We assessed schizotypy, depression, and anxiety as well as healthy and unhealthy behaviours and a wide range of sociodemographic scores using online surveys from residents of Germany and the United Kingdom over 1 year during the COVID-19 pandemic. Four independent samples were collected (April/May 2020: N=781, September/October 2020: N=498, January/February 2021: N=544, May 2021: N=486). The degree of schizotypy was measured using the Schizotypal Personality Questionnaire (SPQ), anxiety, and depression symptoms were surveyed with the Symptom Checklist (SCL-27), and healthy and unhealthy behaviours were assessed with the Coronavirus Health Impact Survey (CRISIS). Structural equation models were used to consider the influence of COVID-19 related concerns and social adversity on depressive and anxiety-related symptoms and schizotypal traits in relation to certain healthy (sleep and exercise) and unhealthy behaviours (alcohol and drug consumption, excessive media use). Results: The results revealed that COVID-19 related life concerns were significantly associated with schizotypy in the September/October 2020 and May 2021 surveys, with anxiety in the September/October 2020, January/February 2021, and May 2021 surveys, and with depressive symptoms in all surveys. Social adversity significantly affected the expression of schizotypal traits and depressive and anxiety symptoms in all four surveys. Importantly, we found that excessive media consumption (>4 hr per day) fully mediated the relationship between COVID-19 related life concerns and schizotypal traits in the January/February 2021 survey. Furthermore, several of the surveys showed that excessive media consumption was associated with increased depressive and anxiety-related symptoms in people burdened by COVID-19 related life. Conclusions: The ongoing uncertainties of the pandemic and the restrictions on social life have a strong impact on mental well-being and especially the expression of schizotypal traits. The negative impact is further boosted by excessive media consumption, which is especially critical for people with high schizotypal traits. Funding: FK received funding from the European Union's Horizon 2020 (Grant number 754,462). SN received funding from the Cundill Centre for Child and Youth Depression at the Centre for Addiction and Mental Health, Toronto, Canada and the Wellcome Trust Institutional Strategic Support Fund from the University of Cambridge.

The 2020 COVID-19 pandemic, and the measures different governments took to contain it, harmed many people's mental well-being. The restrictions, combined with pandemic-related uncertainty, caused many individuals to experience increased stress, depression, and anxiety. Many people turned to unhealthy behaviours to cope, including consuming more alcohol or drugs, using media excessively, developing poor sleeping habits, or reducing the amount of exercise they did. Stress, drugs, poor sleep, and uncertainty can increase an individual's risk of developing psychotic symptoms, including delusions, hallucinations, or difficulty thinking clearly. These symptoms may be temporary or part of a more lasting condition, like schizophrenia. The risk of developing these symptoms increases in people with 'schizotypal traits', such as a lack of close relationships, paranoia, or unusual or implausible beliefs. These individuals may be especially vulnerable to the harmful mental health effects of the pandemic. Daimer et al. demonstrated that people who were more worried about their life stability or financial situation during the 2020 COVID-19 pandemic had worse mental well-being than those who felt secure. In the experiments, volunteers completed a series of online mental health questionnaires at four different time points during the pandemic. People who reported feeling lonely, having negative thoughts, or experiencing fewer positive social interactions had more symptoms of mental illness. People who experienced more life disruptions also reported more anxiety or depression symptoms and more schizotypal traits. Daily consumption of at least four hours of digital media exacerbated negative mental health symptoms, and people with more pandemic-related life concerns also spent more time on digital media Daimer et al. suggest that increased media consumption among people with pandemic-related hardships may have increased mental health symptoms and schizotypal traits in these individuals. The survey results suggest that maintaining a healthy lifestyle, including meaningful relationships, is essential to staying mentally healthy during extreme situations like a global pandemic. Protective interventions ­ such as strengthening social support networks, providing mental health education, or increasing mental healthcare provisions ­ are essential to prevent poor mental health outcomes during future crises.

Optimal surface estimation and thresholding of confocal microscope images of biofilms using Beer's Law.

Beer's Law explains how light attenuates into thick specimens, including thick biofilms. We use a Bayesian optimality criterion, the maximum of the posterior probability distribution, and computationally efficiently fit Beer's Law to the 3D intensity data collected from thick living biofilms by a confocal scanning laser microscope. Using this approach the top surface of the biofilm and an optimal image threshold can be estimated. Biofilm characteristics, such as bio-volumes, can be calculated from this surface. Results from the Bayesian approach are compared to other approaches including the method of maximum likelihood or simply counting bright pixels. Uncertainty quantification (i.e., error bars) can be provided for the parameters of interest. This approach is applied to confocal images of stained biofilms of a common lab strain of Pseudomonas aeruginosa, stained biofilms of Janthinobacterium isolated from the Antarctic, and biofilms of Staphylococcusaureus that have been genetically modified to fluoresce green.

Neural development: The semantics of axon guidance.

Recent studies of the semaphorin family of axon guidance signals and their receptors have revealed a surprising versatility in the ways that they can be used solve problems in neural development, and provided new opportunities for understanding how guidance information is interpreted beneath the cell surface.

A cDNA clone for a polyadenylated RNA-binding protein of Xenopus laevis oocytes hybridizes to four developmentally regulated mRNAs.

Xenopus laevis oocytes contain a unique group of proteins which decrease during oogenesis, bind poly(A) RNA, and possibly play a role in the regulation of translation. A monoclonal antibody generated against one of these proteins was used to screen an expression vector cDNA library. A cDNA clone was isolated and confirmed to code for the binding protein by in vitro translation of hybrid-selected RNA followed by immunoprecipitation. This cDNA, when used in RNA gel blots, hybridized to four transcripts of 2.0, 1.7 (two transcripts of similar size), and 1.2 kilobases. All of the transcripts decreased in amount during oogenesis and were not evident in somatic cells. In addition, the fraction of the transcripts associated with polysomes decreased during oogenesis. Digestion of the cDNA insert with PstI generated two fragments of 220 and 480 base pairs which, when used as probes in an RNA gel blot, hybridized to unique as well as common transcripts. Genomic Southern blots suggested the presence of a single gene, indicating that these transcripts arose by alternative processing.

The Adjustment Disorder--New Module 20 as a Screening Instrument: Cluster Analysis and Cut-off Values.

BACKGROUND: Adjustment disorder (AjD) is a transient mental health condition emerging after stressful life events. Its diagnostic criteria have recently been under revision which led to the development of the Adjustment Disorder--New Module 20 (ADNM-20) as a self-report assessment. OBJECTIVE: To identify a threshold value for people at high risk for AjD. METHODS: As part of a randomized controlled trial evaluating a self-help manual for burglary victims, the baseline data of all participants (n=80) were analyzed. Besides the ADNM-20, participants answered self-report questionnaires regarding the external variables post-traumatic stress disorder symptomatology, depression, anxiety, and stress levels. We used cluster analysis and ROC analysis to identify the most appropriate cut-off value. RESULTS: The cluster analysis identified three different subgroups. They differed in their level of AjD symptomatology from low to high symptom severity. The same pattern of impairment was found for the external variables. The ROC analysis testing the ADNM-20 sum scoreagainst the theory-based diagnostic algorithm, revealed an optimal cut-off score at 47.5 to distinguish between people at high risk for AjD and people at low risk. CONCLUSION: The ADNM-20 distinguishes between people with low, moderate, and high symptomatology. The recommendation for a cut-off score at 47.5 facilitates the use of the ADNM-20 in research and practice.

A new enhancer of position-effect variegation in Drosophila melanogaster encodes a putative RNA helicase that binds chromosomes and is regulated by the cell cycle.

In Drosophila melanogaster, position-effect variegation of the white gene has been a useful phenomenon by which to study chromosome structure and the genes that modify it. We have identified a new enhancer of variegation locus, Dmrnahel (hel). Deletion of mutation of hel enhances white variegation, and this can be reversed by a transformed copy of hel+. In the presence of two endogenous copies, the transformed hel+ behaves as a suppressor of variegation. hel is an essential gene and functions both maternally and zygotically. The HEL protein is similar to known RNA helicases, but contains an unusual variant (DECD) of the DEAD motif common to these proteins. Potential HEL homologues have been found in mammals, yeast and worms. HEL protein associates with salivary gland chromosomes and locates to nuclei of embryos and ovaries, but disappears in mitotic domains of embryos as chromosomes condense. We propose that the HEL protein promotes an open chromatin structure that favors transcription during development by regulating the spread of heterochromatin, and that HEL is regulated by, and may have a role in, the mitotic cell cycle during embryogenesis.

Genetic and developmental characterization of Dmca1D, a calcium channel alpha1 subunit gene in Drosophila melanogaster.

To begin unraveling the functional significance of calcium channel diversity, we identified mutations in Dmca1D, a Drosophila calcium channel alpha1 subunit cDNA that we recently cloned. These mutations constitute the l(2)35Fa lethal locus, which we rename Dmca1D. A severe allele, Dmca1D(X10), truncates the channel after the IV-S4 transmembrane domain. These mutants die as late embryos because they lack vigorous hatching movements. In the weaker allele, Dmca1D(AR66), a cysteine in transmembrane domain I-S1 is changed to tyrosine. Dmca1D(AR66) embryos hatch but pharate adults have difficulty eclosing. Those that do eclose have difficulty in fluid-filling of the wings. These studies show that this member of the calcium channel alpha1 subunit gene family plays a nonredundant, vital role in larvae and adults.

Analysis of the signal-averaged P-wave duration in patients with percutaneous coronary angioplasty-induced myocardial ischemia.

To assess the impact of angioplasty-induced myocardial ischemia on the duration of the surface P wave, patients undergoing elective angioplasty of isolated lesion in the left anterior descending, circumflex or right coronary arteries were monitored with a 3-channel electrocardiographic Holter system. The leads used were modified bipolar chest leads V5, aVF and V2 (CM-V5, CS-aVF and CM-V2). After echocardiographic signal-averaging, the earliest onset and the latest offset of the P wave were identified in all of the above time-aligned signal-averaged leads, and the composite maximal P duration was measured under 10 x magnification. The maximal ST-segment shift during balloon inflation was also measured in all of the above leads at 60 ms after the J point. In the study group comprising 47 patients, the mean signal-averaged P-wave duration was 125.0 +/- 16 ms at baseline versus 130.0 +/- 15 ms during balloon inflation, p less than 0.005. In the left anterior descending coronary artery group (n = 23), the mean signal-averaged P-wave duration was 122.4 +/- 17 ms and 131.3 +/- 16 ms during balloon inflation, p less than 0.005). In the group with a right coronary artery lesion (n = 18), the values were 127.3 +/- 14 ms and 128.4 +/- 13 ms respectively (p = not significant). Significant increases in the P-wave duration were found to occur in groups both with (n = 34) and without (n = 13) ST-segment shift greater than or equal to 1 mm (both p less than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)

Reduction of Culiseta melanura fitness by eastern equine encephalomyelitis virus.

The traditional view of interactions between arboviruses and their arthropod vectors is that vector hosts become increasingly resistant to parasites; parasite attenuation occurs; or through the process of coevolution, resistance and attenuation occur in concert. Detrimental effects from arboviruses are only seen when vector and virus are not yet well adapted. Results from this study indicate that eastern equine encephalomyelitis (EEE) virus reduces survival and reproduction (fitness) of the mosquito Culiseta melanura, which is required for transmission of EEE virus in North America. Mosquito virulence was not measurably attenuated in virus isolates recovered 55 year apart. This virus did not affect the ability of mosquitoes to obtain a blood meal or the rate of mosquito oocyte development. Results from this study support those from earlier investigations with other mosquito-virus interactions and suggest that reproductively successful arboviruses can have detrimental effects on their mosquito vectors.

Pathologic changes in the midgut of Culex tarsalis following infection with Western equine encephalomyelitis virus.

Midguts of two strains of the mosquito vector Culex tarsalis were examined by light and electron microscopy following infection with western equine encephalomyelitis virus. Infection of the highly susceptible Knight's Landing strain with high-titered blood meals resulted in pathologic changes in the midgut epithelium after 2-4 days of incubation; lesions included sloughing of epithelial cells into the lumen and necrosis of cells in situ. Infection of Knight's Landing strain mosquitoes with low-titered blood meals and infection of the less susceptible Fort Collins strain with high-titered blood meals did not result in a significant increase in detached luminal cells, with respect to uninfected controls. Sloughing of infected cells into the midgut lumen may contribute to modulation of the mosquito infection. Lesions in the midgut of Cx. tarsalis are inconsistent with traditional views that regarded arbovirus infections of mosquito vectors as non-pathologic. These findings demonstrate that mosquito pathology is not an oddity limited to the previously described interaction between Culiseta melanura and eastern equine encephalomyelitis virus, and suggest that alphaviruses in general may adversely affect their mosquito vectors in nature.

Detection of eastern equine encephalomyelitis virus deposition in Culiseta melanura following ingestion of radiolabeled virus in blood meals.

We examined deposition of eastern equine encephalomyelitis virus (EEEV) in the alimentary tract of its enzootic mosquito vector, Culiseta melanura, to detect potential sites of initial infection. Artificial viremias were created by injecting purified [3H]uridine-labeled EEEV intravenously into one-day-old chicks. Mosquitoes were allowed to engorge, incubated 1-2 hr, fixed, and whole abdomens and thoraces of 25 specimens embedded. Tagmata were sectioned, mounted on slides and coated with autoradiographic emulsion. Following exposures of 1-4 months, slides were developed and examined microscopically. In the majority of mosquitoes, label (virus) was detected only within the midgut; most virus was concentrated in a band of expressed serum adjacent to the abdominal midgut epithelium. Small amounts of virus were also deposited within folds in the cardial thoracic midgut of 96% of mosquitoes. Viral penetration into epithelial cells was detected throughout the abdominal midgut in all mosquitoes, and in the thoracic midgut of 20%. A small number (4/25) of mosquitoes examined showed signs of leaky abdominal midguts, with virus detected in the abdominal hemocoel. Concentration of EEEV in expressed serum adjacent to the abdominal midgut epithelium may enhance initial midgut infection. Leaky abdominal midguts exhibited by some mosquitoes may also facilitate rapid systemic infections of Cs. melanura. Deposition of virus in thoracic alimentary tissues suggests the possibility of early EEEV infection of the anterior midgut.

Histological parameters useful in the identification of multiple bloodmeals in mosquitoes.

Culex nigripalpus females were given double and, in 2 cases, triple (interrupted) bloodmeals separated by various intervals. Mosquitoes given single meals served as controls. Using Azan-stained serial paraffin sections, we could identify multiple meals separated by 1-72 hr in 44 of 53 cases (84.6%). Among the histological parameters of importance in the identification of double and triple bloodmeals are the peritrophic membrane secreted around each bloodmeal, the plug which forms between the anterior and posterior midgut, the layer of heme which forms as a bloodmeal is digested, and remnants of the pupal meconium and/or pupal-pharate adult peritrophic membranes. The parameters we have identified will help determine the incidence of double bloodfeeding in wild populations which, in turn, should enhance our understanding of the transmission of pathogens by mosquitoes.

The effect of colonization upon aedes aegypti susceptibility to oral infection with yellow fever virus.

Two colonies of Aedes aegypti were established from two independent collections from Vero Beach, Florida. Eleven sequential generations of the first colony were tested for variation in oral susceptibility to infection with yellow fever virus (YFV). Each generation was also assayed for genetic variability at seven enzyme loci using electrophoretic techniques. Significant differences in infection rates were detected between some generations. These differences were significantly correlated with genetic variation at the malate dehydrogenase locus. Seven generations from the second colony were examined simultaneously for variation in susceptibility to YFV. Significant differences were also detected between some of these generations. The results suggest that colonization may have an effect on the genetic and phenotypic variation in a mosquito strain, and that genetically based variation for susceptibility to infection with YFV occurs in populations of Ae. aegypti.