Aedes albopictus oviposits with other Aedes species in artificial oviposition cups: a case study in Knox County,Tennessee, U.S.A.

Interspecific associations between two mosquito species can lead to effects such as competition, species displacement, and species stability. To better understand Aedes albopictus (Skuse) and other Aedes species, we monitored eggs in artificial oviposition cups (ovitraps) within Knox County, TN, U.S.A., during the 2016 and 2017 mosquito seasons. In 2016, one black and one white ovitrap were placed at 18 sites for 21 weeks, while in 2017 black and white ovitraps baited with grass‐infused or deionized water were placed at 11 sites for nine weeks. Eggs were identified to species and resulting counts were used to determine the degree of interspecific association using Cole's coefficients (C₇) and the degree of heterogeneity across space and time using generalized linear mixed models (GLMM). Here, Aedes mosquitoes are generally ovipositing in black cups with grass‐infused water, and Ae. albopictus eggs co‐occurred with other Aedes species more often than would be expected. Finding a positive significant interspecific association between Ae. albopictus and other Aedes eggs suggests that methods used to control Ae. albopictus may also control other Aedes mosquitoes. Finding that Ae. albopictus co‐occurs with other Aedes mosquitoes warrants additional research to evaluate outcomes associated with co‐occurrence within the study area.     

Population genomics of the Asian tiger mosquito,Aedes albopictus: insights into the recent worldwide invasion

Aedes albopictus, the “Asian tiger mosquito,” is an aggressive biting mosquito native to Asia that has colonized all continents except Antarctica during the last ~30–40 years. The species is of great public health concern as it can transmit at least 26 arboviruses, including dengue, chikungunya, and Zika viruses. In this study, using double‐digest Restriction site‐Associated DNA (ddRAD) sequencing, we developed a panel of ~58,000 single nucleotide polymorphisms (SNPs) based on 20 worldwide Ae. albopictus populations representing both the invasive and the native range. We used this genomic‐based approach to study the genetic structure and the differentiation of Ae. albopictus populations and to understand origin(s) and dynamics of the recent invasions. Our analyses indicated the existence of two major genetically differentiated population clusters, each one including both native and invasive populations. The detection of additional genetic structure within each major cluster supports that these SNPs can detect differentiation at a global and local scale, while the similar levels of genomic diversity between native and invasive range populations support the scenario of multiple invasions or colonization by a large number of propagules. Finally, our results revealed the possible source(s) of the recent invasion in Americas, Europe, and Africa, a finding with important implications for vector‐control strategies.     

Assessment of Aedes albopictus (Skuse) (Diptera: Culicidae) clutch size in wild and laboratory populations

Aedes albopictus (Skuse) is an invasive mosquito species found across the southern U.S. with range expansion into many northern states. Intra‐ and interspecific larval competition have been evaluated for Ae. albopictus with respect to subsequent adult size, immature and adult survivability, and its capacity to vector pathogens as an adult. However, limited data are available on egg production as related to larval rearing conditions. Because Ae. albopictus is a container‐inhabiting mosquito that oviposits in resource‐limited habitats, it is found under variable density‐dependent conditions. Therefore, we examined the impact of specific rearing conditions on Ae. albopictus clutch size and adult body size; comparing the egg production values and wing lengths from known developmental densities to those from field‐collected populations. Field populations varied significantly among collection sites in mean clutch size (23 to 46). These clutch sizes were comparable to the mean clutch sizes of females reared at the larval densities of nine (20 eggs) and three (53 eggs) larvae per 3 ml of water in the laboratory. Field populations experienced density‐dependent effects impacting adult mosquito size. Mosquitoes from the four sample sites had mean wing lengths of 1.99, 2.47, 2.51, and 2.54 mm, which were less than the mean wing length of mosquitoes reared at larval densities of three larvae per 3 ml of water (2.57 mm).     

Effect of water depth on predation frequency by diving beetles on mosquito larvae prey

Diving behavior and its frequency may differ among species of mosquito larvae because of differences in predation pressure. The present study aimed to investigate the relationship between water depth and predation frequency on two mosquito species, Culex tritaeniorhynchus (wetland breeder) and Aedes albopictus (container breeder), by the diving beetle Eretes griseus. Culex tritaeniorhynchus spends more time at the surface than A. albopictus, which spends more time thrashing underwater. When intact mosquito larvae of both species were present, the diving beetles consumed almost all A. albopictus larvae (98.3%). After all the A. albopictus larvae had been consumed, the diving beetles began to prey on C. tritaeniorhynchus. In order to compare the effect of position on the predation preference of the diving beetles, equal numbers of both species were heat‐killed and allowed to settle on the bottom of the container. When all the dead mosquito larvae had sunk to the bottom of a plastic container, the diving beetles caught both mosquito species at random. These results indicate that mosquito larvae near the surface were eaten less frequently by diving beetles than those at the bottom. The low diving frequency of C. tritaeniorhynchus is regarded as a form of anti‐predatory behavior.     

Aquatain AMF efficacy on juvenile mosquito stages in control of Culex pipiens complex and Aedes albopictus

Aquatain^® mosquito formulation (AMF) is a silicone-based monomolecular film, which has recently been approved for use in the European Union. The physical mode of action based on lowering water surface tension prevents mosquito larvae/pupae respiration. Additionally, AMF disables gravid females from landing on the water surface and obstructs the natural oviposition process. Due to multistage effects on mosquitoes, AMF could be a product of choice for defined water body and container breeders such as Culex pipiens L. complex, principal vector of West Nile virus in Europe, and the invasive Aedes albopictus (Skuse) (both Diptera: Culicidae), vector of dengue and chikungunya viruses. The primary objectives of this study were to evaluate the efficacy of AMF, to determine the susceptibility of the immature forms of C. pipiens and A. albopictus, and the persistence/longevity of the product to suppress the eclosion of adults. AMF achieved high mortality rates of juvenile A. albopictus and C. pipiens under laboratory conditions. However, in the field C. pipiens larvae showed higher susceptibility to AMF than A. albopictus. Pupae of the two mosquito species were highly susceptible to the presence of AMF. When C. pipiens juveniles were exposed to AMF in the wild, effects lasted for 21 days in densely covered water bodies and 56 days in water recipients with less vegetation. In both breeding sites, natural habitat and artificial water recipient, the two mosquito species with high impact on public health in Europe could successfully be suppressed by application of AMF (1 ml m⁻²).     

Ultrastructure of the fetal membranes of the oviparous kingsnake,Lampropeltis getula (Colubridae) as revealed by scanning electron microscopy

In reptilian sauropsids, fetal (extraembryonic) membranes that line the eggshell sustain developing embryos by providing for gas exchange and uptake of water and eggshell calcium. However, a scarcity of morphological studies hinders an understanding of functional specializations and their evolution. In kingsnakes (Lampropeltis getula), scanning electron microscopy reveals two major fetal membranes: the chorioallantois and yolk sac omphalopleure. In early development, the chorioallantois contains tall chorionic epithelial cells, avascular connective tissue, and enlarged allantoic epithelial cells. During its maturation, the chorionic and allantoic epithelia thin dramatically and become underlain by a rich network of allantoic capillaries, yielding a membrane ideally suited for respiratory gas exchange. Yolk sac development initially is like that of typical lizards and snakes, forming an avascular omphalopleure, isolated yolk mass (IYM), and yolk cleft. However, unlike the situation in most squamates studied, the omphalopleure becomes transformed into a “secondary chorioallantois” via three asynchronous events: flattening of the epithelium, regression of the IYM, and vascularization by the allantois. Progressive expansion of chorioallantois parallels growing embryonic needs for gas exchange. In early through mid‐development, external surfaces of both the chorionic and omphalopleure epithelium show an abundance of irregular surface protrusions that possibly increase surface area for water absorption. We postulate that the hypertrophied allantoic epithelial cells produce allantoic fluid, a viscous substance that facilitates water uptake and storage. Our findings are consistent with a previous study on the corn snake Pantherophis guttatus, but include new observations and novel functional hypotheses relevant to a reconstruction of basal squamate patterns. J. Morphol. 276:1467–1481, 2015. © 2015 Wiley Periodicals, Inc.     

Understanding the mechanism of Chikungunya virus vector competence in three species of mosquitoes

Chikungunya virus (CHIKV) is primarily transmitted by Aedes spp. mosquitoes. The present study investigated vector competence for CHIKV in Aedes aegypti and Aedes albopictus mosquitoes found in Madurai, South India. The role of receptor proteins on midguts contributing to permissiveness of CHIKV to Aedes spp. mosquitoes was also undertaken. Mosquitoes were orally infected with CHIKV DRDE‐06. Infection of midguts and dissemination to heads was confirmed by immunofluorescence assay at different time points. A plaque assay was performed from mosquito homogenates at different time points to study CHIKV replication. Presence of putative CHIKV receptor proteins on mosquito midgut epithelial cells was detected by virus overlay protein binding assay (VOPBA). The identity of these proteins was established using mass spectrometry. CHIKV infection of Ae. aegypti and Ae. albopictus midguts and dissemination to heads was observed to be similar. A plaque assay performed with infected mosquito homogenates revealed that CHIKV replication dynamics was similar in Aedes sp. mosquitoes until 28 days post infection. VOPBA performed with mosquito midgut membrane proteins revealed that prohibitin could serve as a putative CHIKV receptor on Aedes mosquito midguts, whereas an absence of CHIKV binding protein/s on Culex quinquefasciatus midguts can partially explain the non‐permissiveness of these mosquitoes to infection.     

Brief communication: Developmental versus functional three‐dimensional geometric morphometric‐based modularity of the human proximal humerus

The proximal humerus is formed by three secondary ossification centers during the postnatal trajectory of the human infant. The ossification centers later grow into the structures of the articular surface, major tubercle, and minor tubercle. There is a purported functional division between the articular surface and the tubercles, with the articular surface mainly responsible for the range of movement of the shoulder joint, and the tubercles bearing the insertions of the rotator cuff muscles, mainly devoted to securing the joint against humeral displacement. Using three‐dimensional geometric morphometrics, we tested the presence of such developmental and functional divisions in the proximal humerus, applying the RV coefficient of Escoufier to these a priori hypothesized modules. Our results indicate that the proximal humerus might be a generally integrated structure. However, a weak signal for modular configuration was present, with slightly stronger support for the two modules depicting the boundaries between the purported functional regions of the epiphysis: the articular surface and the tubercles. Am J Phys Anthropol 154:459–465, 2014. © 2014 Wiley Periodicals, Inc.     

Morphology of eggs and spermatheca of <Emphasis Type="Italic">Odontotarsus purpureolineatus</Emphasis> (Heteroptera,Scutelleridae)

The morphology of the spermatheca and eggs of Odontotarsus purpureolineatus were studied by optical microscopy and scanning electron microscopy. The spherical eggs were about 1.35 mm long and 1.09 mm wide. The egg batches generally consist of 13–14 eggs. The egg surface is covered by polygonal (hexagonal and pentagonal shapes prevail) ridges and tiny chorionic tubercles. There were 8–10 aero-micropylar processes between the polygons. The spermatheca of O. purpureolineatus is characterized by a spherical spermathecal bulb, a pumping region, a flange of pump and dilation of spermathecal duct. Spermathecal processes and a median spermathecal dilation with sclerotized rod are missing. The spermathecal bulb and the pumping region possess many pores.     

Asian tiger mosquitos (Aedes albopictus) in urban Tokyo,Japan show low cytochrome c oxidase subunit 1 diversity

The Asian tiger mosquito, Aedes albopictus, is a common mosquito in East and Southeast Asia, but its habitats have expanded to the American, African and Australian continents, Europe and many other areas. Aedes albopictus can transmit some important arboviruses that cause human mortality. To control the global spread of this mosquito, genetic analyses of A. albopictus populations have been undertaken throughout the world. In Japan, however, few attempts have been made to characterize the population structures of these mosquitos. In this study, adult A. albopictus populations were sampled from seven parks in the urban area of Tokyo, and analyses of the mitochondrial cytochrome c oxidase subunit 1 (COI) gene base sequences revealed small genetic variations. Only three haplotypes were identified, and most of the samples belonged to a single haplotype. In addition, despite a developed international trade network, no establishment of A. albopictus populations from multiple origins was found. We also evaluated the genetic diversity outside Tokyo using data from a previous study for comparison and found that the genetic diversity in the urban area of Tokyo was lower than that in Nagasaki City (Japan) and in other countries, including the USA and Italy, where A. albopictus populations from Japan have been established.     

The eggshell of the cherry fly Rhagoletis cerasi

One of the major pests in Greek cherry orchards is the cherry fly Rhagoletis cerasi (Diptera: Tephritidae). In order to complete our comparative work on the chorion assembly of other representatives of the fruit flies (e.g. Ceratitis capitata and Dacus oleae) we studied eggshell morphogenesis in the cherry fly. The oocyte is surrounded by several distinct layers which are produced during choriogenesis. The eggshell consists of the vitelline membrane, a fibrous layer of possible water-proofing function, an innermost chorionic layer, endochorionic and exochorionic layers. The endochorion shows a branched configuration with irregular cavities, and the exochorion consists of inner and outer layers for better embryo protection. At the anterior region of the follicle, the hexagonal borders of the follicle cells are created by endochorionic material, covered by both inner and outer exochorion. This area resembles the D. melanogaster chorionic appendages and therefore can serve for plastron respiration. The structural results support the phylogenetic relationships among the tephritids (Rhagoletis is closer to Ceratitis than Dacus). The presence of peroxidase in the endochorion, detected by diaminobenzidine, is consistent with the eggshell hardening at the end of choriogenesis, following the same pattern with the other fruit flies studied so far. Two major chorionic proteins are found both in R. cerasi and in C. capitata and therefore general conclusions can be drawn from this study, concerning the pattern of choriogenesis, which all dipteran insects follow, in order to create a resistant and functional eggshell, and the high conservation of the proteinaceous components of the chorion among species in the order.     

The presence of white eggs in the monitoring of Aedes albopictus ( Diptera: Culicidae) by ovitraps

Using international trading and passive transportation routes, the Asian tiger mosquito, Aedes albopictus (Skuse, 1894), has colonized Europe. While the most common tool to monitor the presence of the mosquito is the ovitrap, other kinds of eggs are occasionally found in the traps as well. Most of the eggs are easy to distinguish, however, some white‐yellow eggs have a similar shape and size to those of the tiger mosquito and are often falsely identified as freshly laid tiger mosquito eggs. We have shown that these eggs had been laid by Psychoda alternata Say, 1824, and the misinterpretation may cause large errors in calculating density and patterns of Ae. albopictus. To avoid mistakes, a microscopic observation should be done at least 48 h after collecting the sample to permit Ae. albopictus eggs to darken.     

Construction of an efficient genomic editing system with CRISPR/Cas9 in the vector mosquito Aedes albopictus

Aedes (Stegomyia) albopictus, also known as the Asian tiger mosquito, is a mosquito which originated in Asia. In recent years, it has become increasingly rampant throughout the world. This mosquito can transmit several arboviruses, including dengue, Zika and chikungunya viruses, and is considered a public health threat. Despite the urgent need of genome engineering to analyze specific gene functions, progress in genetical manipulation of Ae. albopictus has been slow due to a lack of efficient methods and genetic markers. In the present study, we established targeted disruptions in two genes, kynurenine hydroxylase (kh) and dopachrome conversion enzyme (yellow), to analyze the feasibility of generating visible phenotypes with genome editing by the clustered regularly interspaced short palindromic repeats (CRISPR)/ CRISPR-associated protein 9 (Cas9) system in Ae. albopictus. Following Cas9 single guide RNA ribonucleoprotein injection into the posterior end of pre-blastoderm embryos, 30%-50% of fertile survivors produced alleles that failed to complement existing kh and yellow mutations. Complete eye and body pigmentation defects were readily observed in GI pupae and adults, indicating successful generation of highly heritable mutations. We conclude that the CRISPR/Cas9-mediated gene editing system can be used mAe. albopictus and that it can be adopted as an efficient tool for genome-scale analysis and biological study.     

Temporal and spatial habitat preferences and biotic interactions between mosquito larvae and antagonistic crustaceans in the field

Investigations on natural antagonists of mosquito larvae found that micro‐crustaceans (e.g., Cladocera) control mosquito populations under experimental conditions. However, their relevance for mosquito control under field situations remains widely unclear because important information about habitat preferences and time of occurrence of crustaceans and mosquito larvae are still missing. In order to fill this knowledge gap, a field study was undertaken in different wetland areas of Saxony, Germany, in different habitats (i.e., grassland, forest, and reed‐covered wetlands). We found negative interactions between larvae of Ae. vexans and predatory Cyclopoida (Crustacean: Copepoda), which both were dominant during the first two weeks of hydroperiod, at ponds located at grassland habitats. Larvae of Cx. pipiens were spatially associated with competing Cladocera, but they colonized ponds more rapidly. Populations of Cladocera established from the third week of hydroperiod and prevented Cx. pipiens colonization thereafter. Ostracoda were highly abundant during the whole hydroperiod, but their presence was restricted to habitats of reed‐covered wetland at one geographical area. Mosquito larvae hardly occurred at those ponds. In general, we found that ponds at the reed‐covered wetlands provided better conditions for the initial development of crustaceans and hence, mosquito larval colonization was strongly inhibited. Grassland habitat, in contrast, favored early development of mosquito larvae. This study showed that micro‐crustaceans are relevant for mosquito management but their impact on mosquito larvae varies between species and depends on environmental conditions.     

Differentiation of Aedes aegypti and Aedes albopictus (Diptera: Culicidae) with egg surface morphology and morphometrics using scanning electron microscopy

Aedes aegypti and Aedes albopictus are potential arboviral vectors leading to high human fatality worldwide. Efforts in the present study were made to differentiate the eggs of A. aegypti and A. albopictus morphologically and morphometrically using scanning electron microscopy (SEM). Morphometrically, these species’ eggs were 48.48% significantly different of the 33 attributes including egg dimensions, micropylar apparatus, dimensions and density of outer chorionic cells (OCCs), tubercles and width of exochorionic network. In comparison to A. aegypti eggs, A. albopictus eggs were significantly smaller and more tapered at the posterior end; however, the micropylar disc of A. aegypti was wider and had incomplete circular sectors whereas it was a narrower polygon without sectors in A. albopictus. These species were also significantly different with regards to OCC which enclose both large central and small peripheral tubercles. Specifically, the exochorionic networks in A. aegypti were interwoven, reticulated and extensively wide whereas they were narrow, prominent and solid-wall-like in A. albopictus. This feature may strengthen A. albopictus eggs against desiccation, when they are laid in containers. The morphometrical and morphological analysis of the egg’s attributes of A. aegypti and A. albopictus may be helpful in understanding egg biology as well as in species confirmation.     

Decoupling of active and passive reasons for the invasion dynamics of Aedes albopictus Skuse (Diptera: Culicidae): Comparisons of dispersal history in the Apennine and Florida peninsulas

Aedes albopictus is an important vector of several diseases including dengue‐ and Chikungunya fever and is a potential vector of Zika‐fever. The invasion dynamics of Aedes albopictus was reconsidered by comparing the temperature‐related development of the mosquito with the observed real geographical distribution in Florida and in Italy. The potential number of generations and the annual dispersal distances of the mosquito were calculated for the estimates. The estimated total dispersals are 3.6–4.6 km/year/generation in Italy and 4.6–5.3 km/year/generation in Florida, values that are at least five to six times higher than those derived from release and recapture studies and from the previously measured flying distances of female Asian tiger mosquitoes. Subtracting the calculated dispersal distances with the known active dispersal of female Ae. albopictus, the passive dispersal component of the total dispersal distances was found to be 2.8–4.1 km/year/generation in Italy and 3.8–4.8 km/year/generation in Florida. Our results confirm that the active dispersal of female mosquitoes plays a secondary role in determining the rate of areal expansion and, in contrast, passive factors may play a primary role. It was concluded, based on similar average values of the passive dispersal distances of the mosquito in Florida and Italy, that at large spatial scales the anthropogenic component can be well estimated.     

The challenge of invasive mosquito vectors in the U.K. during 2016–2018: a summary of the surveillance and control of Aedes albopictus

Mosquito‐borne diseases resulting from the expansion of two key vectors, Aedes aegypti and Aedes albopictus (Diptera: Culicidae), continue to challenge whole regions and continents around the globe. In recent years there have been human cases of disease associated with Chikungunya, dengue and Zika viruses. In Europe, the expansion of Ae. albopictus has resulted in local transmission of Chikungunya and dengue viruses. This paper considers the risk that Ae. aegypti and Ae. albopictus represent for the U.K. and details the results of mosquito surveillance activities. Surveillance was conducted at 34 points of entry, 12 sites serving vehicular traffic and two sites of used tyre importers. The most common native mosquito recorded was Culex pipiens s.l. (Diptera: Culicidae). The invasive mosquito Ae. albopictus was detected on three occasions in southern England (September 2016, July 2017 and July 2018) and subsequent control strategies were conducted. These latest surveillance results demonstrate ongoing incursions of Ae. albopictus into the U.K. via ground vehicular traffic, which can be expected to continue and increase as populations in nearby countries expand, particularly in France, which is the main source of ex‐continental traffic.     

Modeling a single season of Aedes albopictus populations based on host‐seeking data in response to temperature and precipitation in eastern Tennessee

In the southern Appalachia of the U.S., Aedes mosquitoes maintain and transmit La Crosse virus (LACV) which causes La Crosse encephalitis, a neuroinvasive disease of children. In response to mosquito outbreaks, communities organize prevention, detection, and response measures that are dependent on local characteristics of the mosquito population and the community. Knowing Ae. albopictus is an accessory vector of LACV and a nuisance biter, our objective was to build a system of ordinary differential equations to model dynamics in a single season using our data and readily available environmental variables that can reflect the abundance and activity of Ae. albopictus. Consequently, we built an Ae. albopictus single‐season mathematical model for eastern Tennessee to fit our 2013 mosquito collection data in order to understand the population fluctuations. We included precipitation, temperature, and rate of change of temperature in the model because Aedes mosquitoes oviposit desiccant tolerant eggs with peak activity occurring over 26° C and those data are readily available and used frequently as forecast predictors. Our ordinary differential equation model accurately fits the data and facilitates predictions and better understanding of Ae. albopictus populations in southern Appalachia.     

Human‐aided and natural dispersal drive gene flow across the range of an invasive mosquito

Human‐aided transport is responsible for many contemporary species introductions, yet the contribution of human‐aided transport to dispersal within non‐native regions is less clear. Understanding dispersal dynamics for invasive species can streamline mitigation efforts by targeting routes that contribute disproportionally to spread. Because of its limited natural dispersal ability, rapid spread of the Asian tiger mosquito (Aedes albopictus) has been attributed to human‐aided transport, but until now, the relative roles of human‐aided and natural movement have not been rigorously evaluated. Here, we use landscape genetics and information‐theoretic model selection to evaluate 52 models representing 9240 pairwise dispersal paths among sites across the US range for Ae. albopictus and show that recent gene flow reflects a combination of natural and human‐aided dispersal. Highways and water availability facilitate dispersal at a broad spatial scale, but gene flow is hindered by forests at the current distributional limit (range edge) and by agriculture among sites within the mosquito's native climatic niche (range core). Our results show that highways are important to genetic structure between range‐edge and range‐core pairs, suggesting a role for human‐aided mosquito transport to the range edge. In contrast, natural dispersal is dominant at smaller spatial scales, reflecting a shifting dominance to natural movement two decades after introduction. These conclusions highlight the importance of (i) early intervention for species introductions, particularly those with readily dispersed dormant stages and short generation times, and (ii) strict monitoring of commercial shipments for transported immature stages of Ae. albopictus, particularly towards the northern edge of the US range.     

Comparison of eggshell porosity and estimated gas flux between the brown‐headed cowbird and two common hosts

The brown‐headed cowbird Molothrus ater is a brood‐parasite that lays eggs in nests of a wide range of host species, including the closely‐related red‐winged blackbird Agelaius phoeniceus and the dickcissel Spiza americana. Although cowbird eggs have accelerated development and hatch sooner than similar‐sized host eggs, this development takes place within a thickened eggshell that could impede gas flux to the developing embryo. We tested the hypothesis that the accelerated development of the cowbird embryo relative to hosts is enabled by an increase in eggshell porosity that allows increased fluxes of respiratory gases to and from the developing embryo. We found cowbird eggshell thickness was significantly greater than the eggshells of these two common hosts. Although the number of pores per egg was similar among all three species, the total pore area per egg in cowbirds was significantly greater than that of either host, despite having a smaller eggshell surface area than the red‐winged blackbird. Cowbird egg pore area was 1.9×larger than that of the red‐winged blackbird. Cowbird eggshells had a significantly greater gas flux than those of the red‐winged blackbird and the dickcissel. When conductance was normalized to published values of egg mass, cowbird eggs had a higher mass‐specific conductance than red‐winged blackbird or dickcissel eggs. These results are consistent with the hypothesis that the rapid development of brown‐headed cowbird embryos is facilitated by increased eggshell porosity, and that changes in eggshell porosity represent an adaptation that enables cowbird eggs to hatch earlier than equivalently‐sized host eggs.