Age-dependent Mortality Research Articles

Early mortality saves energy: estimating the energetic cost of excess offspring in a seabird.

Offspring are often produced in excess as insurance against stochastic events or unpredictable resources. This strategy may result in high early-life mortality, yet age-specific mortality before offspring independence and its associated costs have rarely been quantified. In this study, we modelled age-specific survival from hatching to fledging using 24 years of data on hatching order (HO), growth and age of mortality of more than 15 000 common tern (Sterna hirundo) chicks. We found that mortality peaked directly after hatching, after which it declined rapidly. Mortality hazard was best described with the Gompertz function, and was higher with later HO, mainly due to differences in baseline mortality hazard, rather than age-dependent mortality. Based on allometric mass-metabolism relationships and detailed growth curves of starving chicks, we estimated that the average metabolizable energy intake of non-fledged chicks was only 8.7% of the metabolizable energy intake of successful chicks during the nestling phase. Although 54% of hatchlings did not fledge, our estimates suggest them to have consumed only 9.3% of the total energy consumption of all hatched chicks in the population before fledging. We suggest that rapid mortality of excess offspring is part of an adaptive brood reduction strategy to the benefit of the parents.

A new framework for evaluating the health impacts of treatment for Gaucher disease type 1

BackgroundThe Disease Severity Scoring System (DS3) is a validated measure for evaluating Gaucher disease type 1 (GD1) severity. We developed a new framework, consisting of health states, transition probabilities between those states, and preferences for those states (utilities) based on the DS3 to predict long-term outcomes of patients starting treatment. We defined nine mutually exclusive (alive) health states based on three DS3 categories: mild (0 ≤ DS3 ≤ 3.5) without symptoms of bone disease; mild with bone pain, mild with severe skeletal complications (SSC) defined as lytic lesions, avascular necrosis, or fracture; moderate (3.5 < DS3 ≤ 6.5) without SSC; moderate with SSC; marked (6.5 < DS3 ≤ 9.5) without SSC; marked with SSC; severe (9.5 < DS3 ≤ 19) without SSC; and severe with SSC. Health-state transition probabilities and utilities were estimated from a longitudinal sample of patients with GD1 who started enzyme replacement therapy (the DS3 Score Study). Age dependent GD1-specific mortality was derived from published data. We used a Markov state-transition model to illustrate how to estimate time spent in each health state.ResultsThe average predicted utilities for each health state ranged from 0.76 for mild disease with no clinical symptoms of bone disease to 0.52 with severe disease with SSC. Transition probabilities depended on disease severity (DS3 score) at treatment initiation and whether patients had undergone a total splenectomy or had an intact spleen/partial splenectomy prior to starting treatment. Patients who started treatment with intact or residual spleens spent more time in better health states than those who started treatment with total splenectomy.ConclusionsThis new framework, which is based on the DS3, can be used to project the long-term outcomes of GD1 patients starting treatment. The framework could also be used to compare the long-term outcomes of different GD1 treatment options.Trial registrationNCT01136304. Registered: May 31, 2010 (retrospectively registered).

On the effects of changing mortality patterns on investment, labour and consumption under uncertainty

In this paper we extend the consumption–investment life cycle model for an uncertain-lived agent, proposed by Richard (1974), to allow for flexible labour supply. We further study the consumption, labour supply and portfolio decisions of an agent facing age-dependent mortality risk, as presented by UK actuarial life tables spanning the time period from 1951–2060 (including mortality forecasts). We find that historical changes in mortality produce significant changes in portfolio investment (more risk taking), labour (decrease of hours) and consumption level (shift to higher level) contributing up to 5% to GDP growth during the period from 1980 until 2010.

Precommitted, Naive and Sophisticated Retirement Time Decision Making

The optimal retirement time decision is an important aspect when planning for retirement and is on the one side affected by several outside influences and on the other by the way people make decisions. In the literature on decision making, mainly three types of decision makers are distinguished: sophisticated, naive and precommitted. We consider the retirement time decision for all three types and find in a deterministic economy, neglecting investment, that all three agree on the timing of retirement. In a economy, the precommitted and naive retirement decisions are calculated for an age-dependent mortality model. Although the sophisticated decision is solved for a constant mortality intensity only, applying the precommitted one enables us to compare the relevance of two important factors. First, the ability to change retirement time upon financial market movements, i.e. account for the stochastic economy and, second, including a realistic life-time distribution with an age-dependent mortality. A numerical welfare analysis reveals that the first factor is more important for young individuals, whereas the second one has a higher relevance at older ages.

Antibody responses to avian influenza viruses in wild birds broaden with age.

For viruses such as avian influenza, immunity within a host population can drive the emergence of new strains by selecting for viruses with novel antigens that avoid immune recognition. The accumulation of acquired immunity with age is hypothesized to affect how influenza viruses emerge and spread in species of different lifespans. Despite its importance for understanding the behaviour of avian influenza viruses, little is known about age-related accumulation of immunity in the virus's primary reservoir, wild birds. To address this, we studied the age structure of immune responses to avian influenza virus in a wild swan population (Cygnus olor), before and after the population experienced an outbreak of highly pathogenic H5N1 avian influenza in 2008. We performed haemagglutination inhibition assays on sampled sera for five avian influenza strains and show that breadth of response accumulates with age. The observed age-related distribution of antibody responses to avian influenza strains may explain the age-dependent mortality observed during the highly pathogenic H5N1 outbreak. Age structures and species lifespan are probably important determinants of viral epidemiology and virulence in birds.

Temporal Scaling of Age-Dependent Mortality: Dynamics of Aging in Caenorhabditis elegans Is Easy to Speed Up or Slow Down, but Its Overall Trajectory Is Stable.

The dynamics of aging is often described by survival curves that show the proportion of individuals surviving to a given age. The shape of the survival curve reflects the dependence of mortality on age, and it varies greatly for different organisms. In a recently published paper, Stroustrup and coauthors ((2016) Nature, 530, 103-107) showed that many factors affecting the lifespan of Caenorhabditis elegans do not change the shape of the survival curve, but only stretch or compress it in time. Apparently, this means that aging is a programmed process whose trajectory is difficult to change, although it is possible to speed it up or slow it down. More research is needed to clarify whether the "rule of temporal scaling" is applicable to other organisms. A good indicator of temporal scaling is the coefficient of lifespan variation: similar values of this coefficient for two samples indicate similar shape of the survival curves. Preliminary results of experiments on adaptation of Drosophila melanogaster to unfavorable food show that temporal scalability of survival curves is sometimes present in more complex organisms, although this is not a universal rule. Both evolutionary and environmental changes sometimes affect only the average lifespan without changing the coefficient of variation (in this case, temporal scaling is present), but often both parameters (i.e. both scale and shape of the survival curve) change simultaneously. In addition to the relative stability of the coefficient of variation, another possible argument in favor of genetic determination of the aging process is relatively low variability of the time of death, which is sometimes of the same order of magnitude as the variability of timing of other ontogenetic events, such as the onset of sexual maturation.

Life Cycles and Mortality

In this paper we extend the consumption-investment life cycle model for an uncertain-lived agent, proposed by Richard (1974), to allow for flexible labor supply. We further study the consumption, labor supply and portfolio decisions of an agent facing age-dependent mortality risk, as presented by UK actuarial life tables spanning the time period from 1951-2060 (including mortality forecasts). We find that historical changes in mortality produces significant changes in portfolio investment (more risk taking), labour (decrease of hours) and consumption level (shift to higher level), but no explicit effect on consumption growth was found due to the full insurance against loss of life.

Characterization of a Novel Murine Model to Study Zika Virus.

The mosquito-borne Zika virus (ZIKV) is responsible for an explosive ongoing outbreak of febrile illness across the Americas. ZIKV was previously thought to cause only a mild, flu-like illness, but during the current outbreak, an association with Guillain–Barré syndrome and microcephaly in neonates has been detected. A previous study showed that ZIKV requires murine adaptation to generate reproducible murine disease. In our study, a low-passage Cambodian isolate caused disease and mortality in mice lacking the interferon (IFN) alpha receptor (A129 mice) in an age-dependent manner, but not in similarly aged immunocompetent mice. In A129 mice, viremia peaked at ∼107 plaque-forming units/mL by day 2 postinfection (PI) and reached high titers in the spleen by day 1. ZIKV was detected in the brain on day 3 PI and caused signs of neurologic disease, including tremors, by day 6. Robust replication was also noted in the testis. In this model, all mice infected at the youngest age (3 weeks) succumbed to illness by day 7 PI. Older mice (11 weeks) showed signs of illness, viremia, and weight loss but recovered starting on day 8. In addition, AG129 mice, which lack both type I and II IFN responses, supported similar infection kinetics to A129 mice, but with exaggerated disease signs. This characterization of an Asian lineage ZIKV strain in a murine model, and one of the few studies reporting a model of Zika disease and demonstrating age-dependent morbidity and mortality, could provide a platform for testing the efficacy of antivirals and vaccines.

Multiple Metazoan Life-span Interventions Exhibit a Sex-specific Strehler-Mildvan Inverse Relationship Between Initial Mortality Rate and Age-dependent Mortality Rate Acceleration.

The Gompertz equation describes survival in terms of initial mortality rate (parameter a), indicative of health, and age-dependent acceleration in mortality rate (parameter b), indicative of aging. Gompertz parameters were analyzed for several published studies. In Drosophila females, mating increases egg production and decreases median life span, consistent with a trade-off between reproduction and longevity. Mating increased parameter a, causing decreased median life span, whereas time parameter b was decreased. The inverse correlation between parameters indicates the Strehler-Mildvan (S-M) relationship, where loss of low-vitality individuals yields a cohort with slower age-dependent mortality acceleration. The steroid hormone antagonist mifepristone/RU486 reversed these effects. Mating and mifepristone showed robust S-M relationships across genotypes, and dietary restriction showed robust S-M relationship across diets. Because nutrient optima differed between females and males, the same manipulation caused opposite effects on mortality rates in females versus males across a range of nutrient concentrations. Similarly, p53 mutation in Drosophila and mTOR mutation in mice caused increased median life span associated with opposite direction changes in mortality rate parameters in females versus males. The data demonstrate that dietary and genetic interventions have sex-specific and sometimes sexually opposite effects on mortality rates consistent with sexual antagonistic pleiotropy.

Use of serologic tests to predict resistance to Canine distemper virus–induced disease in vaccinated dogs

The objective of the current study was to determine whether detection of Canine distemper virus (CDV)-specific serum antibodies correlates with resistance to challenge with virulent virus. Virus neutralization (VN) assay results were compared with resistance to viral challenge in 2 unvaccinated Beagle puppies, 9 unvaccinated Beagle dogs (4.4-7.2 years of age), and 9 vaccinated Beagle dogs (3.7-4.7 years of age). Eight of 9 (89%) unvaccinated adult dogs exhibited clinical signs after virus challenge, and 1 (13%) dog died. As compared to adult dogs, the 2 unvaccinated puppies developed more severe clinical signs and either died or were euthanized after challenge. In contrast, no clinical signs were detected after challenge of the 9 adult vaccinated dogs with post-vaccination intervals of up to 4.4 years. In vaccinated dogs, the positive and negative predictive values of VN assay results for resistance to challenge were 100% and 0%, respectively. Results indicate that dogs vaccinated with modified live CDV can be protected from challenge for ≤4.4 years postvaccination and that detection of virus-specific antibodies is predictive of whether dogs are resistant to challenge with virulent virus. Results also indicate that CDV infection in unvaccinated dogs results in age-dependent morbidity and mortality. Knowledge of age-dependent morbidity and mortality, duration of vaccine-induced immunity, and the positive and negative predictive values of detection of virus-specific serum antibodies are useful in development of rational booster vaccination intervals for the prevention of CDV-mediated disease in adult dogs.

A theoretical model of the evolution of actuarial senescence under environmental stress

Free-living organisms are exposed to a wide range of stressors, all of which can disrupt components of stress-related and detoxification physiology. The subsequent accumulation of somatic damage is widely believed to play a major role in the evolution of senescence. Organisms have evolved sophisticated physiological regulatory mechanisms to maintain homeostasis in response to environmental perturbations, but these systems are likely to be constrained in their ability to optimise robustness to multiple stressors due to functional correlations among related traits. While evolutionary change can accelerate due to human ecological impacts, it remains to be understood how exposure to multiple environmental stressors could affect senescence rates and subsequently population dynamics and fitness. We used a theoretical evolutionary framework to quantify the potential consequences for the evolution of actuarial senescence in response to exposure to simultaneous physiological stressors – one versus multiple and additive versus synergistic – in a hypothetical population of avian “urban adapters”. In a model in which multiple stressors have additive effects on physiology, species may retain greater capacity to recover, or respond adaptively, to environmental challenges. However, in the presence of high synergy, physiological dysregulation suddenly occurs, leading to a rapid increase in age-dependent mortality and subsequent population collapse. Our results suggest that, if the synergistic model is correct, population crashes in environmentally-stressed species could happen quickly and with little warning, as physiological thresholds of stress resistance are overcome.

Identifiability problem for recovering the mortality rate in an age-structured population dynamics model

In this article is studied the identifiability of the age-dependent mortality rate of the Von Foerster–Mc Kendrick model, from the observation of a given age group of the population. In the case where there is no renewal for the population, translated by an additional homogeneous boundary condition to the Von Foerster equation, we give a necessary and sufficient condition on the initial density that ensures the mortality rate identifiability. In the inhomogeneous case, modelled by a non-local boundary condition, we make explicit a sufficient condition for the identifiability property, and give a condition for which the identifiability problem is ill-posed. We illustrate this latter case with numerical simulations.

Gender and age dependent mortality from nervous diseases in Azerbaijan

To assess age- and sex-related changes in the mortality from nervous diseases at the population level. Methods of descriptive statistics and analysis of qualitative traits were applied. We analyzed 13580 medical certificates of cause of death from nervous diseases (all classes of ICD-10). The mortality rate varied with age, the main trend of which was the dynamic growth. Age-specific mortality rates for men and women differed from each other: in most ages (20-24, 30-34, 45-49, 50-54, 55-59, 65-69), the likelihood of mortality was higher in men, and at the age of 5-9, 15-19, 60-64, 70 and more years in women. After the standardization of gender differences by age, the mortality risk of nervous illnesses disappeared (146.74 and 144.16 per 100 thousand for men and women, respectively). There were significant differences in the proportion of nervous diseases of all-cause mortality among the population in the groups stratified by age and sex. It is believed that situational factors is a cause of actual prevailing of gender age- and sex-related mortality risks. Gender features of age-related risk of mortality from nervous diseases are characterized by the multidirectional dynamics of likelihood of mortality and specific weight of nervous diseases among all causes of mortality. The actual gender features of age-related risk of mortality from nervous diseases are generally caused by situational factors (different age structure and unequal level of the general mortality among male and female population) which disappear after standardization.

A new cecal slurry preparation protocol with improved long-term reproducibility for animal models of sepsis.

Sepsis, a life-threatening systemic inflammatory response syndrome induced by infection, is widely studied using laboratory animal models. While cecal-ligation and puncture (CLP) is considered the gold standard model for sepsis research, it may not be preferable for experiments comparing animals of different size or under different dietary regimens. By comparing cecum size, shape, and cecal content characteristics in mice under different experimental conditions (aging, diabetes, pancreatitis), we show that cecum variability could be problematic for some CLP experiments. The cecal slurry (CS) injection model, in which the cecal contents of a laboratory animal are injected intraperitoneally to other animals, is an alternative method for inducing polymicrobial sepsis; however, the CS must be freshly prepared under conventional protocols, which is a major disadvantage with respect to reproducibility and convenience. The objective of this study was to develop an improved CS preparation protocol that allows for long-term storage of CS with reproducible results. Using our new CS preparation protocol we found that bacterial viability is maintained for at least 6 months when the CS is prepared in 15% glycerol-PBS and stored at -80°C. To test sepsis-inducing efficacy of stored CS stocks, various amounts of CS were injected to young (4–6 months old), middle-aged (12–14 months old), and aged (24–26 months old) male C57BL/6 mice. Dose- and age-dependent mortality was observed with high reproducibility. Circulating bacteria levels strongly correlated with mortality suggesting an infection-mediated death. Further, injection with heat-inactivated CS resulted in acute hypothermia without mortality, indicating that CS-mediated death is not due to endotoxic shock. This new CS preparation protocol results in CS stocks which are durable for freezing preservation without loss of bacterial viability, allowing experiments to be performed more conveniently and with higher reproducibility than before.

Increased coagulation and suppressed generation of activated protein C in aged mice during intra-abdominal sepsis.

Sepsis is a life-threatening clinical condition that is particularly serious among the elderly who experience considerably higher mortality rates compared with younger patients. Using a sterile endotoxemia model, we previously reported age-dependent mortality in conjunction with enhanced coagulation and insufficient levels of anti-coagulant factor activated protein C (aPC). The purpose of the present study was to further investigate the mechanisms for age-dependent coagulation and aPC insufficiency during experimental sepsis. Intra-abdominal sepsis was induced by cecal ligation and puncture (CLP) using 21 or 16 gauge (G) needles (double-puncture) on young (4 to 6 mo old) and aged (20 to 25 mo old) male C57BL/6 mice. When compared with young mice, aged mice showed significantly increased mortality (92% vs. 28%), systemic inflammation, and coagulation in the lung and kidney after 21G CLP. Young mice with more severe CLP (16G) showed a mortality rate and inflammation equivalent to aged mice with 21G CLP; however, enhanced coagulation and kidney dysfunction were significant only in the aged. In young mice, increased levels of aPC after CLP were coupled with reduced levels of protein C (PC), suggesting the conversion of PC to aPC; however, PC and aPC levels remained unchanged in aged mice, indicating a lack of PC to aPC conversion. Activation of fibrinolysis, determined by plasma d-dimer levels, was similar regardless of age or CLP severity, and plasminogen activator inhibitor-1, an inhibitor of fibrinolysis, showed severity-dependent induction independent of age. These results suggest that enhanced coagulation in aged mice during sepsis is due to dysfunction of the PC activation mechanism.

Differentiation of neurons restricts Arbovirus replication and increases expression of the alpha isoform of IRF-7.

Susceptibility to alphavirus infection is age dependent, and host maturation is associated with decreased virus replication and less severe encephalitis. To identify factors associated with maturation-dependent restriction of virus replication, we studied AP-7 rat olfactory bulb neuronal cells, which can differentiate in vitro. Differentiation was associated with a 150- to 1,000-fold decrease in replication of the alphaviruses Sindbis virus and Venezuelan equine encephalitis virus, as well as La Crosse bunyavirus. Differentiation delayed synthesis of SINV RNA and protein but did not alter the susceptibility of neurons to infection or virion maturation. Additionally, differentiation slowed virus-induced translation arrest and death of infected cells. Differentiation of uninfected AP-7 neurons was associated with changes in expression of antiviral genes. Expression of key transcription factors was increased, including interferon regulatory factor 3 and 7 (IRF-3 and IRF-7) and STAT-1, suggesting that neuronal maturation may enhance the capacity for antiviral signaling upon infection. IRF-7 produced by undifferentiated AP-7 neurons was exclusively the short dominant negative γ-isoform, while that produced by differentiated neurons was the full-length α-isoform. A similar switch in IRF-7 isoforms also occurred in the brains of maturing C57BL/6J mice. Silencing of IRF expression did not improve virus multiplication in differentiated neurons. Therefore, neuronal differentiation is associated with upregulation of transcription factors that activate antiviral signaling, but this alone does not account for maturation-dependent restriction of virus replication. Viral encephalomyelitis is an important cause of age-dependent morbidity and mortality. Because mature neurons are not readily regenerated, recovery from encephalitis suggests that mature neurons utilize unique antiviral mechanisms to block infection and/or clear virus. To identify maturational changes in neurons that may improve outcome, we compared immature and mature cultured neurons for susceptibility to three encephalitic arboviruses and found that replication of Old World and New World alphaviruses and a bunyavirus was reduced in mature compared to immature neurons. Neuronal maturation was associated with increased baseline expression of interferon regulatory factor 3 and 7 mRNAs and production of distinct isoforms of interferon regulatory factor 7 protein. Overall, our studies identified maturational changes in neurons that likely contribute to assembly of immunoregulatory factors prior to infection, a more rapid antiviral response, increased resistance to virus infection, and improved survival.

A low-cost mesocosm for the study of behaviour and reproductive potential in Afrotropical mosquito (Diptera: Culicidae) vectors of malaria.

A large-scale mesocosm was constructed and tested for its effectiveness for use in experiments on behaviour, reproduction and adult survivorship in the Afrotropical malaria vector Anopheles gambiae s.s. Giles (Diptera: Culicidae) in temperate climates. The large space (82.69 m(3) ) allowed for semi-natural experiments that increased demand on a mosquito's energetic reserves in an environment of widely distributed resources. A one-piece prefabricated enclosure, made with white netting and vinyl, prevented the ingress of predators and the egress of mosquitoes. Daylight and white materials prompted the mosquitoes to seclude themselves in restricted daytime resting sites and allowed the easy collection of dead bodies so that daily mortality could be assessed accurately using a method that accounts for the loss of a proportion of bodies. Here, daily, age-dependent mortality rates of males and females were estimated using Bayesian Markov chain Monte Carlo simulation. In overnight experiments, mosquitoes successfully located plants and took sugar meals. A 3-week survival trial with a single cohort demonstrated successful mating, blood feeding, oviposition and long life. The relatively low cost of the mesocosm and the performance of the mosquitoes in it make it a viable option for any behavioural or ecological study of tropical mosquitoes in which space and seasonal cold are constraining factors.

Time-evolution of age-dependent mortality patterns in mathematical model of heterogeneous human population

The widely-known Gompertz law of mortality states the exponential increase of mortality with age in human populations. Such an exponential increase is observed at the adulthood span, roughly after the reproductive period, while mortality data at young and extremely old ages deviate from it. The heterogeneity of human populations, i.e. the existence of subpopulations with different mortality dynamics, is a useful consideration that can explain age-dependent mortality patterns across the whole life-course. A simple mathematical model combining the heterogeneity of populations with an assumption that the mortality in each subpopulation grows exponentially with age has been proven to be capable of reproducing the entire mortality pattern in a human population including the observed peculiarities at early- and late-life intervals. In this work we fit this model to actual (Swedish) mortality data for consecutive periods and consequently describe the evolution of mortality dynamics in terms of the evolution of the model parameters over time. We have found that the evolution of the model parameters validates the applicability of the compensation law of mortality to each subpopulation separately. Furthermore, our study has indicated that the population structure changes so that the population tends to become more homogeneous over time. Finally, our analysis of the decrease of the overall mortality in a population over time has shown that this decrease is mainly due to a change in the population structure and to a lesser extent to a reduction of mortality in each of the subpopulations, the latter being represented by an alteration of the parameters that outline the exponential dynamics.

Sex-specific patterns of aging in sexual ornaments and gametes.

Sex differences in age-dependent mortality and reproductive success are predicted to drive the evolution of sexually dimorphic patterns of reproductive investment over life. However, this prediction has not been fully explored because it is difficult to measure primary and secondary sexual traits over the life spans of males and females. Here we studied a population of fowl, Gallus gallus, to gain longitudinal data on a sexual ornament (the comb), quantity of gametes produced, and gamete quality (sperm velocity and egg mass) of males and females. Our results reveal pronounced differences between the sexes in age-specific patterns of reproductive investment. In males, comb size decreased linearly with age, high sperm quality early in life was associated with reduced sperm quality late in life, and high sperm production was related to early death. In contrast, female comb size and egg mass were maximized at intermediate ages, and fecundity was independent of life span. Finally, the way traits were related in males did not change over life, whereas in females the association between fecundity and comb size changed from positive to negative over the lifetime of a female, indicating that aging may lead to trade-offs in investment between traits in females. These results show that males and females differ in reproductive investment with age, in terms of both the expression of individual traits and their phenotypic covariance.

Reduced survival and reproductive success generates selection pressure for the dengue mosquito Aedes aegypti to evolve resistance against infection by the microsporidian parasite Vavraia culicis

The success and sustainability of control measures aimed at reducing the transmission of mosquito-borne diseases will depend on how they influence the fitness of mosquitoes in targeted populations. We investigated the effects of the microsporidian parasite Vavraia culicis on the survival, blood-feeding behaviour and reproductive success of female Aedes aegypti mosquitoes, the main vector of dengue. Infection reduced survival to adulthood and increased adult female mosquito age-dependent mortality relative to uninfected individuals; this additional mortality was closely correlated with the number of parasite spores they harboured when they died. In the first gonotrophic cycle, infected females were less likely to blood-feed, took smaller meals when they did so, and developed fewer eggs than uninfected females. Even though the conditions of this laboratory study favoured minimal developmental times, the costs of infection were already being experienced by the time females reached an age at which they could first reproduce. These results suggest there will be selection pressure for mosquitoes to evolve resistance against this pathogen if it is used as an agent in a control program to reduce the transmission of mosquito-borne human diseases.