Correlating Feather Structure, Wettability, and Robustness with Ecological Behavior of Aquatic Birds By Jesus 0. Guardado Submitted to the Department of Materials Science and Engineering in Partial Fulfillment of the Requirements for the Degree of ARCBHVES Bachelor of Science at the MASSACHUSETTS INSTITUTE OF TECHNOLOGY Massachusetts Institute of Technology NOV 10 2015 June 2011 LIBRARIES 0 2011 Jesus 0. Guardado. All rights reserved. The author hereby grants to MIT permission to reproduce and to distribute publicly paper and electronic copies of this thesis document in whole or in part in any medium now known or hereafter created. Signature of Author.......................................... Signature redacted Departmrof44p'rials Science and Engineering May 6, 2011 Certified by.................................................... redacted 'Signature Snrec Robert E. Cohen St. Laurent Professor of Chemical Engineering Theis'Spervisor Certified by......................................Signafure redacted Michael F. Rubner TDK Professor of Materials Science and Engineering Course III ThesifrS ervisor Accepted by.......................... .Signature redacted Lionel C. Kimerling Thomas Lord Professor of Materials Science and Engineering Chairman, Undergraduate Thesis Committee This page was intentionally left blank. Correlating Feather Structure, Wettability, and Robustness with Ecological Behavior of Aquatic Birds By Jesus 0. Guardado Submitted to the Department of Materials Science and Engineering on May 6, 2011 in Partial Fulfillment of the Requirements for the Degree of Bachelor of Science in Materials Science and Engineering ABSTRACT In nature, aquatic birds can interact with water without their feathers being easily wetted; some species dive tens of meters and emerge to spread their wings to dry. In past studies attempting to connect such ecological behavior and feather structure, the typical approach of microscopy has demonstrated the difficulty in characterizing specimens as delicate and complex as feathers by visual techniques alone. In this work, the question was addressed of how various species balance the wettability problem with the need to dive to various depths or to remain on or near the water surface as dictated by their feeding habits. Texture of wing feathers from six different species of aquatic birds was characterized by measuring contact angles and applying the previously developed framework of the effective spacing ratio, D*, and robustness factor, A *, according to the Cassie-Baxter relation for composite interfaces. This "effective microscopy" technique was successfully employed to assess the wettability and robustness of bird feather textures. The observable water-related behaviors of diving, wing-spreading, shallow foraging, and dabbling for the species studied were explained as partly determined by feather structure, exhibiting effectiveD* analysis as an adequate technique for characterizing complex, textured surfaces, fabricated or natural. Thesis Supervisor: Professor Robert E. Cohen Title: St. Laurent Professor of Chemical Engineering Course III Thesis Supervisor: Professor Michael F. Rubner Title: TDK Professor of Materials Science and Engineering 3 This page was intentionally left blank. 4 Table of Contents 1 2 LIST OF FIGURES ............................................................................................................................. LIST OF TABLES ............................................................................................................................... ACKNOW LEDGEM ENTS...................................................................................................................8 6 7 INTRODUCTION .............................................................................................................................. 9 BACKGROUND .............................................................................................................................. 11 2.1 ECOLOGICAL BEHAVIOR OF INTEREST: BIRD INTERACTIONS WITH FLUIDS ................................................. 2.1.1 2.1.2 PROFILES OF AQUATIC BIRD SPECIES STUDIED .................................................................................. 2.2 2.2.1 2.2.2 2.2.3 2.2.4 Phalacrocoracidae: cormorants and shags..................................................................... Anhinga rufa: African darter ......................................................................................... Tadorna tadorna: common shelduck.............................................................................. Anas platyrhynchos: mallard......................................................................................... 2.3 EFFECTIVE MICROSCOPY TO CHARACTERIZE SURFACES IN TERMS OF D .................................................. 2.4 W ETTABILITY, COMPOSITE INTERFACES, AND TEXTURED SURFACES ........................................................ 11 14 16 18 18 20 21 21 22 24 Cassie-Baxter (CB) relation and D*................................................................................ Robustness parameter, A*, and breakthrough pressure, Pb............................................ 25 27 2.5 IMPACT OF OPENNESS OF THE W EAVE ON SURFACE WETTABILITY.......................................................... 29 2.6 IMPACT OF OPENNESS OF THE WEAVE ON BREAKTHROUGH PRESSURE, P6 ........................ 31 2.4.1 2.4.2 3 Past attempts to characterize feather topography ........................................................ Difficulties of characterizing topography of bird feathers ............................................. . . . .. . . . . . . . . . .. . . . . . . . . EXPERIM ENTAL PROCEDURES ...................................................................................................... BIRD FEATHER SPECIMENS ........................................................................................................... COATING M ETHODOLOGY............................................................................................................ 3.1 3.2 3.2.1 3.2.2 3.3 Fluorodecyl POSS (polyhedral oligomeric silsesquioxane) and Tecnoflon *1 solution ...... Coating sm ooth and textured surfaces......................................................................... SURFACE CHARACTERIZATION ...................................................................................................... 3.3.1 3.3.2 Contact angle measurements ........................................................................................ Scanning electron microscopy (SEM).............................................................................. RESULTS........................................................................................................................................ 4 34 34 35 35 36 37 37 37 38 4.1 APPARENT CONTACT ANGLES FOR BIRD FEATHERS FROM EACH SPECIES................................................... 38 4.2 "EFFECTIVE MICROSCOPY" OF FEATHERS WITH D*............................................................................. 40 4.3 SCANNING ELECTRON MICROSCOPY (SEM ) OF SELECTED FEATHERS ...................................................... 43 4.4 CRITICAL DIVING DEPTH FROM ROBUSTNESS PARAMETER, A , AND BREAKTHROUGH PRESSURE, Ps................. 47 5 DISCUSSION..................................................................................................................................48 5.1 APPARENT CONTACT ANGLES USED TO ASSESS WETTABILITY OF BIRD FEATHERS ........................................ 48 5.2 EFFECTIVE MICROSCOPY (1-D MODEL) CHARACTERIZES FEATHER TEXTURE ............................................. 49 5.3 EFFECTIVE D* IS BETTER INDICATOR OF FEATHER WETTABILITY THAN D* FROM GEOMETRY ............................. 50 5.4 ROLE OF WETTABILITY (O Aov) AND FEATHER TEXTURE (D*) ON ECOLOGICAL BEHAVIOR.............................. 51 5.5 ROLE OF ROBUSTNESS FACTOR, A*, AND BREAKTHROUGH PRESSURE, PB, ON ECOLOGICAL BEHAVIOR................ 54 6 CONCLUSIONS .............................................................................................................................. 56 7 FUTUR E W ORK ............................................................................................................................. 57 8 REFERENCES ................................................................................................................................. 59 9 APPENDICES ................................................................................................................................. 61 9.1 APPENDIX A: M ATLAB* CODE FOR D* FITTING PROCEDURE ............................................................... 61 9.2 APPENDIX B: SLIDING ANGLES MEASURED FOR SELECTED BIRDS AND LIQUIDS.............................................. 62 5 List of Figures FIGURE 1. BIRD FEATHERS INTERACTING W ITH LIQUIDS ......................................................................... 11 FIGURE 2. EFFECT OF SURFACE CHEMISTRY ON THE WETTABILITY OF A FLAT SURFACE BY WATER AND OCTANE ....... 13 FIGURE 3: ECOLOGICAL BEHAVIORS OF INTEREST OF AQUATIC BIRDS.........................................................14 FIGURE 4. ATTEMPT TO CHARACTERIZE FEATHERS BY D* = (r + d) / r VALUES VISUALLY OBTAINED VIA MICROSCOPY .......................................................................................................................................... 15 FIGURE 5. SEM MICROGRAPHS (100X MAGNIFICATION) OF REED CORMORANT FEATHERS FROM THE SIX DIFFERENT PRIMARY AND BODY CONTOUR FEATHER CATEGORIES.......................................................................16 FIGURE 6. GENERALIZED NON-WETTING DIAGRAM FOR TEXTURED SURFACES: DIP-COATED, WOVEN MESHES........23 FIGURE 7. SCHEMATIC DIAGRAMS OF THE INTERFACE FORMED BY A WATER DROPLET .................................. FIGURE 25 8. SCHEMATIC DIAGRAM OF REPRESENTATIVE D REGIMES FOR A PARALLEL ARRAY OF CYLINDERS............27 FIGURE 9. EFFECT OF LENGTH SCALE, R, ON THE ROBUSTNESS OF A TEXTURED SURFACE .............................. 29 FIGURE 10. M ODEL OF BIRD FEATHER W ETTED BY OILS ........................................................................... 31 FIGURE 11. M ODEL OF BIRD FEATHER SUBMERGED IN WATER...................................................................33 FIGURE 12. PHOTOGRAPHS OF WING, BREAST, AND BELLY FEATHERS FOR THE SIX BIRD SPECIES.......................35 FIGURE 13. CHEMICAL STRUCTURE OF FLUORODECYL POSS AND TECNOFLON* FLUOROELASTOMER.................36 FIGURE 14. GENERALIZED NON-WETTING DIAGRAM FOR DIP-COATED BIRD FEATHERS FROM EACH SPECIES...........41 FIGURE 15. SCANNING ELECTRON MICROGRAPHS FOR TOPOGRAPHY OF WING FEATHERS OF AFRICAN DARTER AND COM M ON SHELDUCK ........................................................................................................... 47 FIGURE 16. MATLAB* CODE USED TO ESTIMATE THE DIMENSIONLESS SPACING RATIO, D*, FOR EACH BIRD SPECIES .......................................................................................................................................... 6 61 List of Tables TABLE 1. A LIST OF THE SIX BIRD SPECIES STUDIED, INCLUDING SCIENTIFIC AND COMMON NAMES AND METRICS FOR DIVING BEHAVIO R ................................................................................................................ 18 TABLE 2. A CATALOG OF THE TWELVE BIRD FEATHER SPECIMEN SUPPLIED BY DR. ANDREW PARKER (BMNH, LONDON), CONTAINING SCIENTIFIC NAMES, COMMON NAMES, AND IDENTIFICATION CODES...................34 TABLE 3. AVERAGE, APPARENT ADVANCING AND RECEDING CONTACT-ANGLE DATA (O*ADv AND O REc, RESPECTIVELY) FOR WING FEATHERS OF EACH SPECIES AND FOR CHEMICALLY IDENTICAL, FLAT, FLUORODECYL POSS SURFACE 39 (0E) W ITH VARIOUS LIQUIDS................................................................................................... SPECIES..................................................42 TABLE 4. EFFECTIVE SPACING RATIO, D , VALUES FOR EACH BIRD TABLE 5. AVERAGE, APPARENT ADVANCING AND RECEDING CONTACT-ANGLE DATA (e*ADV AND 0 *REC, RESPECTIVELY) WITH VARIOUS LIQUIDS FOR WING FEATHERS OF THE SPECIES WITH HIGHEST AND LOWEST D , AFRICAN DARTER AND COMMON SHELDUCK, RESPECTIVELY. FOR EACH SPECIES, LIQUIDS ARE LISTED IN ORDER OF DECREASING SURFACE TENSION................................................................................................................ TABLE 6. RGEOMETRY CALCULATED AND D 42 AS MEASURED FROM SCANNING ELECTRON MICROGRAPHS AND CORRESPONDING, FOR BARBS AND BARBULES OF AFRICAN DARTER AND COMMON SHELDUCK FEATHERS47 DGEOMETRY GEOMETRY TABLE 7. COMPILATION OF FITTED D , CALCULATED A , EXPECTED PB, EQUIVALENT BREAKTHROUGH SWIMMING VELOCITY, AND BREAKTHROUGH DEPTH FOR EACH BIRD SPECIES.......................................................48 TABLE 8. EFFECTIVE D VALUES (D*EFF) FROM THIS WORK AND D = (R + D) / R VALUES FROM THE LITERATURE ....................................................................................................................................... 50 TABLE 9. CATALOG FOR THE SIX SPECIES OF METRICS OF INTEREST FOR THIS STUDY FOR ANALYZING BIRD-WATER INTERACTIONS. SPECIES ARE ARRANGED IN ORDER OF INCREASING D, DECREASING A, AND DECREASING, PREDICTED HB- ----------. -- - - - -- - - --............................................................................................... 52 TABLE 10. SLIDING ANGLE (() MEASUREMENTS FOR WATER AND ETHYLENE GLYCOL ON WING FEATHERS FROM SPECIES IN THE PHALACROCORACIDAE FAMILY . .......................................................................... 7 62 Acknowledgements I thank God for an able mind, which He has caused to give Him thanks. I thank my advisor, Prof Robert Cohen, for accepting me as an undergraduate student in his lab and being so supportive and amiable. I also thank Prof Michael Rubner for his help throughout, from my introduction into to the Cohen lab and even before, and for accepting me as my Course 3 advisor. I thank my graduate student advisor, Shreerang Chhatre, for working so closely with me and always taking the time to clarify and explain things, and through whom I have increasingly grown to appreciate the ways a researcher and engineer approaches problems. From the Cohen lab, I thank Siddarth Srinivasan for all his technical help, in particular with SEM images, goniometer troubleshooting, and general conversations. Also from the Cohen lab, I thank Jonathan DeRocher for good lab safety and generally helping keep things operating and Adam Meuler for insightful conversations about my project and broader topics. I thank Dr. Andrew Parker for his collaboration and provision of the bird feathers-and for his hospitality during my time in Oxford. Thank you also to Dr. Joseph M. Mabry from the Edwards Air Force Base, CA for his contribution of the POSS materials. I thank Daisy, my wife-to-be, for reminding me of God's sovereignty and what truly matters. I thank my parents, brother, and sister, for their love and labor in general and specifically during my four years at MIT. I thank my younger sister, who gladly proofread my papers since I was in high school and was spared from doing the same for my thesis. Finally, I thank MIT and the Department of Materials Science and Engineering and its people for an enriching four years. 8 1 Introduction In nature, aquatic birds interact with water with ease, and their feathers are not wetted; ducks are a prime example. Remarkably, some aquatic bird species dive tens of meters and emerge to spread their wings to dry. These behaviors and other bird interactions with water are of great interest. To better understand these behaviors, bird-feather topography is examined. The typical approach of microscopy in past studies has shown the topography of feathers is difficult to characterize because they make for delicate, fluffy specimens with structure too complex to characterize by microscopy alone. In this project, the texture of feathers belonging to several aquatic birds from six different species was studied. This thesis is broadly divided into two sections. For the first section of this study, a single, dimensionless parameter (D*) that characterizes surface texture-effectively acting as a "microscope"- and was obtained for each species using contact angle measurements and the Cassie-Baxter relationship for textured surfaces. Complementarily, resistance against water penetration was subsequently computed. In previous work in the Cohen group, this resistance to wetting has been characterized in terms of the breakthrough pressure or the robustness parameter (A *), and in this work, the threshold for non-wetting was also defined by calculating A * for wing feathers from each species. As the contrast across past studies reveals, even after natural specimens are characterized, difficult questions of function often remain. For the second section of this thesis, the effective spacing ratio, D*, and the robustness factor, A *, served to correlate feather structure with the ecological behavior of these aquatic birds, specifically their diving depths and the wingspreading behaviors some exhibit for drying under sunlight. Beyond a particular threshold depth, we expect water to wet bird feathers. 9 Consequently, the question is addressed of how the various species of birds balance the wettability problem with the need to dive to various depths or to remain on or near the surface as dictated by their feeding habits. In addition to diving, other water-related behavior of aquatic birds such as wing-spreading, shallow foraging, and dabbling were linked to the calculated metrics, namely the effective spacing ratio, robustness factor, and breakthrough depth. The "effective microscopy" technique was successfully employed to correlate bird feather texture with wettability and robustness, and to elucidate the role feather texture plays in the waterrelated behavior of aquatic birds. 10 2 2.1 Background Ecological Behavior of Interest: Bird Interactions with Fluids Plumage encloses an insulating layer of air close to the bird's body and ensures adequate thermoregulation. Aquatic birds, then, require highly water-repellent, or hydrophobic, feathers to prevent water from penetrating into the air layer. Bird feathers are naturally water-repellent due to the surface chemistry and structure of their feathers, which along with behavior help counter the effects of hydrostatic pressure. The base component of feathers, the fibrous, structural protein keratin, is inherently water-repellent, and the natural oils the uropygial glands of birds produce and secrete, help repel water. Smeared throughout the feathers during preening, oils from the uropygial glands have been shown to chiefly function to maintain supple and strong feathers and not primarily to repel water [1]. Figure 1 emphasizes how water easily flows off the back of an aquatic bird, but oil does not. The increased wettability of bird feathers by oils is an unfortunate vulnerability often made most visible by ecological disasters such as oil spills, like the crude-oil leak in 2010 from a BP drilling rig in the Gulf of Mexico. Water (y7,= 72.1 mN/m) (d) (c) (b) (a) Figure 1. Bird feathers interacting with liquids. A healthy duck (a) interacting with water, which beads and forms relatively high contact angles on the surface of aquatic bird feathers (b) compared to oil that can come into contact with a bird's coat (c), soiling it by fully wetting the plumage. (d) Rapeseed oil as an example of lower-surface-tension liquids, which more easily wet a bird feather surface. Water colored with methylene blue and rapeseed oil colored with oil red. Images adapted from http://griffyclan007.wordpress.com/2010/06/21/bp-oil-spill-disaster, http://www.conservationinstitute.org, and [2], adapted with permission. 11 Surface chemistry affects a surfaces ability to repel liquids. Wettability, or the degree to which a liquid wets a surface, is determined by the contest between intermolecular, cohesive forces within the liquid and interfacial, adhesive forces between liquid and solid-surface molecules. The chemical constituents of the liquid and the solid surface both matter. Strictly considering surface chemistry, if a glass slide were coated with nature's keratin, for example, a water droplet on such a surface would form a contact angle in the range of 80 to 1000 [3]. As Figure 1 and Figure 2 illustrate, oils and other lower-surface-tension liquids more easily wet such a surface and display a smaller contact angle, which is the angle between the line tangent to the curved liquid/vapor interface and the solid surface Figure 2 shows how adhesive interactions between a liquid droplet and a flat, solid surface can dramatically vary based on the solid-surface energy, which is dictated by surface chemistry. A water droplet and an octane droplet are placed on flat, chemically different surfaces with decreasing surface tensions. In this example, octane was selected as a representative hydrocarbon and one of the main components of gasoline rather than treating the surface with many different oils. For the same surface, a water droplet will display consistently higher contact angles than an oil droplet due to the oil's lower surface tension and, thus, relatively weaker cohesive interactions and stronger adhesive interactions (Figure 2a). Whether with a fluorodecyl POSS or keratin surface, for instance, this result is observed. Moreover, when surface chemistry is changed, the contact angle formed (shown as a red line tangent to the curved air/liquid interface in the images in Figure 2) decreases monotonically for both water and octane with decreasing solid-surface energy (Figure 2b and Figure 2c). Surface chemistry alone, however, plays a limited role in achieving high contact angles on a flat surface. Even coatings with low solid-surface energies may offer a mix of results due to specific interactions resulting from non-ideal polar, and nonpolar (dispersion) hydrogen- 12 bond-donating, and hydrogen-bond-accepting contributions to the solid-surface energy by the compounds on a surface, as previous work characterizing the low-solid-surface-energy coating utilized in this thesis has shown [4]. Along with chemical composition, the other major contribution to liquid repellency comes from structure, or topography, the determining factor for whether water droplets will flow off a feather surface or wet and penetrate between the barbs and barbules. (c) Octane droplet (a) Oil droplet on... Water droplet on... OE, water (0) 180 Air oane (0) 180 Air Fluorodecyl POSS Poly(dimethyl siloxane). PDMS Keratin 110 Poly(methyl methacrylate). PMMA 70 65 -- Fluorodecyl POSS Hydrophilic polymers _- Clean glass 0 <10 -0 PMMA / PDMS. Keratin Figure 2. Effect of surface chemistry on the wettability of a flat surface by water and octane. (a) Advancing contact angles formed on flat surfaces (OE) of different chemical composition by water (liquid-surface tension, y1v = 72.1 mN/m) and octane (yIv = 21.6 mN/m) droplets. A surface of keratin, a basic component of bird feathers, gives lower contact angles than a surface coated with fluorodecyl POSS, a very low-surface-tension molecule (ys, ~ 10 mN/m). (b) OE values of a water droplet on different flat surfaces are greater than the OE values formed by (c) octane on the same coated, flat surfaces. Adapted from [5]. 13 2.1.1 Pastattempts to characterizefeather topography Deep-diving birds, such as cormorants, shags, and other aquatic birds routinely dive up to tens of meters for food and are known to subsequently dry their wings by spreading them in sunlight for extended periods of time (Figure 3). Noting these behaviors, researchers have attempted to correlate it to the structure of bird feathers, with notable efforts for the cormorant and darter, but there is a lack of consensus [6-8]. Particularly, in a 1968 study, Rijke correlated the two by reporting on the wing-spreading behaviors of cormorants and studying feather barbs (c) (b) (a) Figure 3: Ecological behaviors of interest of aquatic birds. (a) A great cormorant with a successful capture. (b) a cormorant diving, likely while foraging, after which it will likely emerge to dry its wings (c) in sunlight while in a wing-spreading posture. Images from (a) http://ibc.lynxeds.com/photo/great-cormorant-phalacrocorax-carbo/fishing-doha-habour, (b) http://commons.wikimedia.org/wiki/File:Cormorantdiving for-foodinMorroBay.jpg, (c) http://tolweb.org/Phalacrocoracidae/26338. and barbules, which he characterized by applying an idealization of cylinders of radius R and inter-cylinder spacing 2D and the dimensionless spacing ratio, D* = (R + D) / R , a model used for describing textiles (Figure 4) [6,8]. Rijke thought in terms of D*, as schematically presented in Figure 4a; to study the feather structure of various species, he employed optical microscopy and photography to measure barb spacing, 2D, and diameter, 2R, from which he calculated D* values for various species (Figure 4b), including four of those also studied in this present work. In his study of breast feathers from terrestrial families and fully aquatic families, Rijke also 14 identified the two areas of interest, water repellency (wettability) and resistance to water penetration (robustness), noting bird feathers of terrestrial birds were generally less wettable, whereas feathers of aquatic birds were generally more robust. One of his main conclusions, that D* for the feathers is correlated to diving, swimming, and wing-spreading behavior, was refuted within one-and-a-half decades by Elowson, who specifically claimed wing-spreading posture (b) (a) Species Water r Air Iwo10 Mallard (A nas platyrhynchos) African Shelduck (Tadorna cana) Reed Cormorant (Phalacrocorax africanus) Bank Cormorant (P. neglectus) Cape Cormorant (P. capensis) Great Cormorant (P. carbo) African Darter (Anhinga rufa) 5.9 5.8 4.3 4.5 4.4 4.8 4.5 . (c) (r + d)/r Figure 4. Attempt to characterize feathers by D* =(r + d) /r values visually obtained via microscopy. (a) Schematic diagram of cross-section of feather barbs modeled as cylinders (barbules not shown) covered by a water droplet of much larger size; r is radius of barbules, d is half-length between barbules. (b) Rijke's spacing-ratio data for various bird species. (c) Photomicrographs representative of breast feathers from two different species of dippers (Cinclidae) examined by Rijke. Scale bar, 300 m. [8], [9] adapted with permission. is independent of D* based on comparisons of contact angle measurements and estimations of D* using scanning electron microscopy (SEM) [10]. Elowson manually measured R and D values from such micrographs to calculate D*, finding poor agreement between measured contact angles and expected contact angles back-calculated from the D* values obtained from the simple 1 -D textile model. Subsequently, Rijke presented a rebuttal to Elowson's objections, specifically 15 addressing the validity of the textile model and the general applicability of the Cassie-Baxter relation derived from physico-chemical principles [11]. Most recently, in 2010, he and collaborator Jesser reapplied the D* = (R + D) / R to another aquatic bird family, employing digital photography, similar to previous studies [9]. The photomicrographs of breast feathers of dippers (Cinclidae) in Figure 4c come from that latest study and typify the problematic complexity and fragility of feather topography, even as probed by recent approaches. They also hint at the shortcomings of techniques that primarily rely on visual measurement to characterize such complex textures. 2.1.2 Difficulties of characterizingtopography of birdfeathers As evidenced by the literature history, the complicated structure of feather texture makes uncertain D* values obtained solely by visual measurement and fails to capture the overall feather topography. An excerpt from several scanning electron micrographs from Elowson's results, Figure 5 exemplifies the complexity of feather structure, and the consequent difficulty in characterizing it. Feather structure is typically hierarchical. The features of feather structure are most generally: a main shaft (rachis), barbs (ramus) that branch out of the main shaft, and Figure 5. SEM micrographs (100x magnification) of reed cormorant feathers from the six different primary and body contour feather categories. Images capture the main shaft or axis (rachis), the barbs or branches (ramus), and the barbules, or minute filaments that branch out in dendritic fashion from the barbs. The feather categories, from left to right, top to bottom: DOV, distal primary outer vane; DIV, distal primary inner vane; BK, back; POV, proximal primary outer vane; PIV, proximal primary inner vane; and BR, breast. [10] 16 barbules that extend from the barbs and often form an interlocking microstructure. Barbules compose the majority of the surface area in these bird feathers. In many cases, because of the intricate feather topography and the small area probed by microscopy, Elowson was forced to report R and D from different areas on a feather as he established D* visually. Elowson's work also dealt with several feather categories, including those coming from the wing, breast, and other areas of the bird's body, as the acronyms within the panels of Figure 5 describe. This broad approach makes sense considering the diversity of nature, even when just considering the expected differences between body feathers and flight feathers, for example. However, for the present study, feathers samples all come from wings, selected because they are more structurally ordered and because the focus on diving and wing-spreading made wing feathers the most relevant choice for both phenomena. Subsequent works by other researchers have also dealt with bird diving behavior, buoyancy, and the wettability of the plumage as a whole, including considerations of energetic consequences and heat losses [12,13], yet these approaches focus even less on characterizing actual feather structure. Even for the purposes of comparing wettability, which is most commonly done with contact angle values, the contact angle formed on a chemically equivalent, flat surface is not accurately known. More specifically, no approach to-date has considered an "effective D*" as obtained through the method employed in this thesis to characterize feather texture. There seems to be a connection between D* and ecological behavior, but the connection as of yet is unclear. We seek to elucidate the correlation between the details of feather texture and the behavioral response of birds. 17 2.2 Profiles of Aquatic Bird Species Studied In this project, the texture of feathers from several aquatic birds from six different species was studied. The wing feathers studied came from six different species of aquatic birds, as tabulated in Table 1 (see also Section 3). Three of the species were chosen from the same Phalacrocoracidaefamily: the reed cormorant, great cormorant, and European shag. The rest of the species each come from distinct families and were chosen because of what is known from observation about their diving and feather-drying behavior. These three other species are: the African darter, common shelduck, and mallard. Table 1. A list of the six bird species studied, including scientific and common names and metrics for diving behavior. Common name Scientific name Divig deth ivin sped Wng- References for: Diving speed spading diving, wingDiig depth spreading Reed cormorant Phalacrocorax africanus Great cormorant Phalacrocoraxcarbo European shag Phalacrocorax aristotelis African darter Anhinga rufa Common shelduck Tadorna tadorna 5 - 6* 0.7-0.85 Y [13], [6,10] 4.7, < 10" 1.1-2.1 Y [14], [6] 33 -35 1.7-1.9 Y [15], [6] Y [16], [10] N [10] <5 Dabbling 0.19 0.16t Dabbling Mallard Anas platyrhynchos Dabbling Dabbling N [10] * from neutral buoyancy experiments, not natural observation; 7 horizontal traveling speed, not diving speed; I usually < 10 m, but can dive to depths of 35 m [17]; Wing-spreading: Y = predictably, N = never 2.2.1 Phalacrocoracidae: cormorants and shags The Phalacrocoracidaefamily is subsumed under the broad Pelecaniformes order and consists of several large water birds with long bodies, long necks, and webbed feet. They possess iridescent-dark back plumage and light or dark front plumage, varying from species to species. Their wingspans are commonly in the range of 3 to 5 ft (0.9 to 1.5 m) [18]. Phalacrocoracidae contains the cormorants and shags, which are seen worldwide and exist in greatest diversity in tropical and temperate zones. They inhabit marine as well as inland waters, 18 and species can be migratory or sedentary. Their wing morphology makes them agile fliers over short distances [17]. Those in marine habitats feed primarily on fish, whereas the diets of inland birds can include fish, frogs, aquatic insects, and water snakes. Their foraging generally relies on underwater food supplies, and cormorants and shags dive, catch a fish with their mouths, and hold it in a pouch akin to that of the pelican. Phalacrocoraxare excellent swimmers. The birds surface dive and use their feet to propel themselves into the air before turning to dive head-first (Figure 3b). The pre-dive leap supplies considerable momentum, and after submergence, the bird continues its dive by simultaneous, feet kicks [19]. After water activities, these birds are known for their wingspreading behavior by which they hold their wings extended and dry their feathers [6,10]. Phalacrocoraxafricanus, or the reed cormorant, is so named due to its location; it is a native of regions throughout Africa, mainly inland [20]. The diving behavior of this bird is not well-documented, and the one study with definitive numbers on diving depth comes from neutral buoyancy experiments with submerged carcasses rather than from natural tracking or observation [13]. The great cormorant (Phalacrocoraxcarbo) is relative large in size-largest of the six North American cormorants, for instance-and in distribution. The species is the most broadly distributed of the cormorants and nearly cosmopolitan, occupying the northwest-Atlantic coasts and also breeding in Europe, Asia, Africa and Australia. It typically nests on cliff ledges and feeds in sheltered, inshore waters. Like others in its family, the great cormorant is a fast flier over short distances at speeds of about 50 km/hour and up to 93 km/hour. This bird will typically dive to depths less than 10 m and captures fish at shallow water less than 20 m deep, but because it feeds primarily on bottom-living fish, it does surface dive up to 35 m [17]. 19 The phalacrocoraxaristotelisresembles cormorants, except it is smaller and slightly slimmer. As its name implies, the European shag is distributed throughout western Europe, usually found along rocky, marine coastlines and islands, rarely traveling far from its breeding area. Like cormorants, European shags feed mainly on fish, although it preferentially consumes different types of fish than the great cormorant, even when they co-occur. Shags forage deeper than great cormorants and notably exhibit the characteristic "leap" out of the water before plunging for prey [21]. 2.2.2 Anhinga rufa: African darter Like the Phalacrocoracidaefamily, the Anhingidae family has been classified under the same broad order of Pelecaniformes. Similar in appearance and comparable in size to cormorants, the Anhinga rufa possesses a distinctly long neck and largely black plumage with white streaks. Distributed throughout sub-Saharan Africa near large bodies of water, these aquatic birds are largely sedentary but are given over to infrequent, opportunistic, local travels in response to environmental conditions. The darter prefers shallow and inland bodies of water and usually avoids marine regions. It primarily consumes fish and is notorious for its low buoyancy, which aids it in its foraging of fish, mostly done in shallow water. In fact, the African darter is colloquially referred to as "snakebird," reflecting how it looks in water, where only its neck protrudes above the water. They are specialist shallow-water divers, and researchers have observed their dives in various ecological conditions, with typical water depths <0.5 m [16]. Notably, as Rijke points out, they can emerge from water "dripping wet" and are able to immediately take up flight. This bird is also known to exhibit spread-wing postures [22]. 20 2.2.3 Tadorna tadorna: common shelduck Tadorna tadorna, or common shelduck, belongs in the Anatidae family, which also includes geese and swans. It is likened to a short-necked goose in appearance and has a long, broad body; it is a mainly white duck with chestnut brown patches, a prominent red bill, blackgreen head, long legs and pink feet, and chestnut and white upper parts. Shelducks are found mainly in coastal areas, although they can also be found around inland waters, and favor saline, muddy habitats. They breed in temperate Eurasia and are seasonally migratory, with the exception of some sedentary, European population [23]. The common shelduck is particularly common around the greater part of Great Britain's and Ireland's coastlines. They feed predominantly on salt-water mollusks and other aquatic invertebrates [23]. The shelduck is better classified as a dabbling duck than a diving duck, meaning it obtains its food on land or just on the water's surface, either by surface dipping or upending (immersing the entire front half of its body in the water so that its hind parts are thrust in the air and its tail remains above the surface). The young dive freely, especially when faced with danger, but the adults only do so when frightened or injured. 2.2.4 Anas platyrhynchos: mallard Also in the Anatidae family, the mallard (Anasplatyrhynchos) shares some qualities with the common shelduck. With regards to appearance, the male is also quite noticeable, possessing an iridescent green head, rusty chest, and green body. In size, however, it is smaller than the common shelduck. This very familiar duck can be seen throughout North America and Eurasia in wetland habitats of all kinds. They migrate southward to warmer regions following the breeding season. It feeds primarily on vegetable matter, insects, worms, and other aquatic invertebrates [24]. 21 Like the shelduck, the mallard is also classified as a dabbling duck. It filter-feeds on the surface of the water and upends in shallow water. Occasionally, though, it will also dive in deeper water [24,25]. Neither the common shelduck nor the mallard display spread-wing postures. 2.3 Effective microscopy to characterize surfaces in terms of D* To elucidate the underlying reasons for these bird behaviors, particularly for bird-water interactions, the details of feather structure serve as this study's starting point. The focus was on quantitatively characterizing feather structure using "effective D*" analysis and the robustness parameter, A *, which we successfully show capture topographical features otherwise lost when characterized directly, visually, through microscopy. With a focus on superhydrophilic, superhydrophobic, and oleophobic surfaces, the Cohen group in collaboration with the McKinley group (Course II) and the Air Force Research Laboratory has developed a design chart framework to predict the wettability by a liquid of a textured surface. In doing so, they address among other problems the question of how stably a textured surface can resist wetting by a contacting liquid. D* and A * analysis requires contact angle measurements not just for water, but for a series of liquids, which include several lower surface tension liquids. However, these liquids more easily wet bird feathers, precluding contact angle measurements. Dip-coating the feathers overcomes that hurdle by essentially making the feathers oleophobic, or oil-repellent, with the additional benefit of ensuring chemically identical surfaces across feather samples, thus excluding chemical variation as an explanation for differences in wettability and robustness. A collaboration with the Air Force Research Lab has granted access to the fluorinated polyhedral oligomeric silsesquioxane (fluoro-POSS or F-POSS) compound. As a hydrophobic crystalline 22 solid with one of the lowest solid-surface energy values reported to-date, F-POSS enables completion of the analysis with a whole array of liquids thanks to the low surface energy of fluoro-POSS molecules and the surface texture formed by the feather's barbs. Fluorodecyl POSS is currently the molecule of choice for designing super-nonwetting surfaces and has proven suitable for the simple dip-coating process, which has been used by the Cohen group to confer flexible, conformal coatings to an array of surface textures [2,26,27]. (b) mesh 50, R = 114 pm (a) 0 dv (0) ' I 0 60 90 120 ' 180 1.0 1- U mesh 50 iAA mesh 100 0.5 1- A mesh 325 60 cos0* =-1+--[sin OE+(-OE)Cos D U) (c) mesh 100, R = 57 pm E] 0.0 0 0 D- -0.5 2 D =2245 120 0.20 (d) -1.0' -1I .0 -0.5 0.0 mesh 325, R = 18 pm 180 1 .0 0.5 cos 0adv Figure 6. Generalized non-wetting diagram for textured surfaces: dip-coated, woven meshes. (a) Cosine of the advancing contact angle on the textured, dip-coated meshes (6*adv) is plotted versus cosine of the advancing contact angle on flat silicon wafers (OE) spin-coated with the same solution as the dip-coated meshes. Diagram contains data for three meshes (b - d) of different length scales, R, and constant spacing ratio, D* = 2.2, each probed with an extensive set of polar and non-polar liquids. The effective spacing ratio, D*, inset in (a) is computed from regression to the advancing contact angle data on the textured (O*adv) and flat (OE) surfaces according to the equation also inset in (a). [28] 23 The Cohen group has applied the "effective D*" approach to fabrics [2,26], wire meshes [28], and other re-entrant texture surfaces including electrospun fiber mats and fabricated microhoodoo arrays [29,30]. Figure 6 highlights the role of surface topography in textured surfaces, for which this single, effective spacing parameter, or effective D*, can be computed and used to quantify the surface's "openness." Distinct from past approaches, this "openness of the weave" is quantified by an effective D*, which is obtained by measuring contact angle measurements on the textured feather surface and on a chemically identical, flat surface. An effective D* is computed for each texture from regression to the advancing contact angle data on the textured surface (O*adv) and on the flat surface (OE) according to the modified Cassie-Baxter relation (Equation 1). The contact angle quantities 0* and OE are explicitly defined in Section 2.4, and the Cassie-Baxter relation is explained in Section 2.4.1. Figure 6 shows three dip-coated, woven meshes that despite having varying length scales, R, share the same effective spacing ratio, D*= 2.2. Their shared openness of the weave as characterized by the effective D* approach is evidenced by their excellent agreement with the Cassie-Baxter model, as shown in the generalized non-wetting diagram for textured surfaces, which plots the cosine 6*adv against the cosine of OE (Figure 6a). This whole procedure was also carried out for bird feathers to study their structure with a particular interest in wettability. 2.4 Wettability, composite interfaces, and textured surfaces As with the structure or roughness of surfaces in general, the structure of feathers can reveal much about their wettability. Wettability is commonly quantified by placing a liquid on a surface and, after it attains thermodynamic equilibrium, measuring this equilibrium contact angle, OE, established between the liquid and the flat, solid surface. Figure 7a illustrates precisely this case of a flat, smooth surface. The equilibrium contact angle is governed by the balance 24 between interfacial tension components and given by Young's relation: cos OE s(yv - V) /v The interfacial tension, y, between solid, liquid, and vapor phases is accounted for and denoted by subscripts s, 1, and v, respectively. Wettability of a material has been shown to depend on two main factors: surface chemistry and topography [31-35]. 2.4.1 Cassie-Baxter (CB) relation and D* The concept of wettability is not limited to flat surfaces; hence its dependence on topography and application to surfaces such as bird feathers. When a liquid droplet interacts with a textured solid surface, it either fully wets the solid, resulting in a Wenzel state [31], or forms a composite solid-liquid-air interface due to air trapped between surface asperities supporting the contacting liquid, producing a "Cassie-Baxter" interface (Figure 7b) [32]. Multivalued surface topography for which a vector normal to the x-y plane (plane running parallel with the surface) intersects the texture at multiple points, also known as re-entrant curvature, has been identified in previous work as essential for supporting a composite interface with lowsurface-tension liquids [29,30]. (a) (b) ]- cylinders 2I) 2R Figure 7. Schematic diagrams of the interface formed by a water droplet on (a) a flat, smooth surface with y indicating the surface tension resulting from interactions between the solid (s), liquid (1), and vapor (v) phases and OE indicating the equilibrium contact angle measured using contact-angle goniometry; and on (b) a rough or textured surface. 25 No longer the equilibrium contact angle (OE) considered for a flat, smooth surface (Figure 7a), the apparent contact angle (0*) formed by the liquid droplet at the composite interface can be calculated using the classical Cassie-Baxter (CB) relation: cos 0* = r#, cos represents roughness of the wetted area and E s -1 , where ro #, the fraction of the projected area of the solid surface in contact with the liquid [34]. The literature as well as previous research in Prof. Cohen's group catalogs many approaches to manipulating the wettability of surfaces, resulting in superhydrophilic (apparent contact angle 0*~ 00), superhydrophobic (0'> 1500), and oleophobic surfaces [29]. Many textured surfaces, including all the wing feather samples considered for this project, can be geometrically modeled as cylindrical arrays, which in turn can be described with radius R and inter-cylinder spacing 2D (Figure 8). This geometrical idealization results in a more conveniently expressed CB relation [2,28,30,36] 1 D cos 0* =1-+ - where r = (z - OE) / sin0, [(r -OE)COSEsinE (1) = R sin OE /(R +D), and D* =(R + D) / R . These equalities show the dependence of the surface texture variables, r, and #, on the equilibrium contact angle (OE), whereas the dimensionless spacing ratio (D*) is independent of the properties of the contacting liquid and, as a purely geometric parameter, quantifies the openness of the surface. The generalized non-wetting diagram in Figure 8 presents various D* regimes for a textured surface modeled as an array of parallel cylinders. Given a uniform R among cylinders, D* =1 describes touching, parallel cylinders, whereas D* = 271 refers to cylinders farther apart. As applied to the bird feathers in this study, length scale, R, is reasonably approximated as constant along the barbule length, and differences in the D* parameter then only reflect variations in spacing between barbules. As a dimensionless parameter, D* is independent of 26 length scale. However, changes in length scale, R, are important for wettability, and its effect on resistance against liquid penetration is discussed in Section 2.4.2 (particularly detailed in Figure 9), which takes robustness of the interface into account. 120 180 D*=I 90 0 60 =x/2 --- 0.5 0 (") 60 -- d= , 0.0 90 8 - 0 (") -0.512 -0.5 0.0 0.5 COSO 180 1.0 * -1.0 -1.0 Figure 8. Schematic diagram of representative D* regimes for a parallel array of cylinders. Supposing negligible variation for R, a dimensionless spacing ratio of unity (D* = 1) refers to the case when the cylinders are touching each other; D* = 2a represents a case when the parallel cylinders are farther apart. 2.4.2 Robustness parameter, A *, and breakthroughpressure, Pb The CB relation only applies when a solid-liquid-air composite interface is assured, a condition threatened by the sagging of the liquid-air interface formed by the trapped air, as the derivation of the CB relation assumes a flat liquid-air interface. If sufficiently severe, the sagging of the liquid-air interface will touch the lower level of solid surface, thus transitioning to a fully wetted state. Robustness against wetting, or the ability to withstand sagging of the liquidair interface, can be quantified by the dimensionless robustness factor, A* [29,30]. The robustness factor is essentially the ratio of the threshold pressure difference that triggers the transition ("breakthrough pressure," Pb) to the characteristic reference pressure, Prf = 2Y7, / where fcap is the capillary length (,,p =, Y cap, / pg ), ylv is the liquid surface tension, and pg is the 27 specific weight. For an arrangement of cylinders with radius R and inter-cylinder spacing 2D, the robustness factor can be calculated from the following expression [2,28,30]: A b (COSOE) cap - Prf R(D*-l) L(D*-1+2sinOE)_ (2) Thus, A * accounts for the equilibrium contact angle (OE), the effective spacing ratio (D*), and the ratio of capillary length (f,) to the feature length scale composing the solid's surface texture (R), f, / R. Values of A * - 1 identify surface-liquid combinations for which a drop will spontaneously transition to a fully-wetted state. All these parameters can be combined in a "design chart" for liquid wettability, in which OE is plotted versus D*, and contours of the apparent contact angle (0*) can be generated from Equation 1 to identify a priorithe combinations of OE and D* that will yield a desired 0* (Figure 9). The developed design chart framework allows for the design of robust interfaces. For example, to design a super-nonwetting, textured surface, 6E and D*values for coordinates above the 0* = 1500 contour would be sought, but robustness must also be considered. Importantly, incorporating the robustness factor, A , into the design chart then predicts the available parameter space for textured surfaces that will maintain robust solid-liquid-air interfaces for a designated threshold A * (commonly A * = 1) below which the surface will fully wet. This region of instability is shown as the grey area in the modified design charts in Figure 9. The shaded grey area diminishes and the unshaded, white area expands from Figure 9a to Figure 9b, highlighting the relevance of R for optimizing the openness of the surface: all other things being equal, having a small feature length scale, R, allows for a high breakthrough pressure, Pb. 28 (b) (a) R cap 120 - A*> I=-> ro (= =1000 100 - - =120" -=1500 80 C"p R 120 0 - posite interface 100 Sca = 2mm, R =2pm, ""a =10 fcap = 2mm, R=20pm, 80 160 _,60 40 A*<1 = composite interface is 20 unstable against any pressure 40 20 perturbation 0/ 1 0 2 4 3 5 6 1 4 3 2 5 6 D D Figure 9. Effect of length scale, R, on the robustness of a textured surface. The modified version of the non-wetting design chart, which predicts the range of allowed D* and OE for forming a robust composite interface, in this case for two values of the ratio of the capillary length (f,) to the feature length scale, fe, / R. Typical surfaces of (a) commercial textile, with R ~ 200 tm, and (b) electrospun mat, R~ 2m. For both surfaces, f, ~ 2 mm. Smaller R, and therefore maximum openness of surface, is desirable to achieve non-wetting, robust interfaces (A* b 1). [28] 2.5 Impact of openness of the weave on surface wettability Probing a surface with various liquids is not just a part of the technique for extracting the D* parameter and quantifying the openness of the weave. Varying the surface tension of a liquid interacting with a textured surface is the equivalent of varying the Laplace pressure or an externally applied pressure. By substituting pressure with surface tension, keeping surface structure constant throughout, Equation 2, Pf = 2y,v / e, and a,,p = 4, / pg show how varying the surface tension, Ylv, alone changes f, and Pef values, resulting in different Pb and A * values. Determined by the liquid used, surface tension, then, can be varied in place of pressure: probing a textured surface with one liquid and then with another of lower surface tension is analogous to 29 submerging that surface-a mesh or feather, for example-in the first liquid, and then lowering the sample deeper to a certain depth still within the same liquid. Under normal conditions, bird feathers normally interact with just water. In this project, bird feathers were exposed to various liquids with different surface tensions to properly characterize the surfaces through D* and A * and to find the expected breakthrough pressures, which should affect bird/water interactions. This concept is well illustrated by subjecting wire meshes to the same testing, as done in previous work in the Cohen group (Figure 10) [28]. As the non-wetting design chart shows, for wire meshes with constant coating chemistry (OE) and feature length scale (R), apparent contact angle increases monotonically with D*. For the parameter space in which a robust composite interface is ensured (A*> 1, white area), a constant R and increasing D* means larger spacing between wires (D), a lower wetted fraction of the solid, and thus higher apparent contact angles. In contrast, when a robust interface is not ensured (A * < 1, shaded grey area), as D* increases, inter-wire spacing D increases, except in this case the air-liquid interface more severely sags and easily transitions to the fully wetted state (Figure 10). In the example in Figure 10 of wire meshes dip-coated with 50% POSS-50% Tecnoflon, the D* spacing of 2.2 preserves a composite interface against the lowered surface tension from contacting water (yiv = 72.1 mN/m) to rapeseed oil (yiv = 35.5 mN/m), but another mesh with D* = 5.1 has a weave that is actually too open and cannot withstand the decreased surface tension when probed with a droplet of rapeseed oil. The air-liquid interface sags to the point of fully wetting (open, right-pointing triangle enclosed in a red circle within the shaded, grey area in Figure 10). The open symbols represent droplets in the fully wetted state. These results convey the connection between wettability and robustness. If a bird feather had been designed like the 30 mesh of smallest D* value, it would more robustly resist wetting, even against the oils that typically wet bird feathers. D*= 2.2, R = 83 gm Water 120 100 Rapeseed oil 80 -exadecane D*= 3.9, R = 83jm ALA 0 60 Heptane 40 Pentane D*= 5.1,R = 83 tm 20 0 1 2 5 4 3 6 D Figure 10. Model of bird feather wetted by oils. Example of design chart for liquid wettability demonstrating effect of varying surface tension. Contact angle data for nine different liquids on three meshes with different D* values and dip-coated with 50% POSS-50% Tecnoflon. Filled symbol indicates a robust composite interface formed by a liquid droplet, whereas open symbol indicates a fully wetted interface formed by a droplet. Left inset image shows non-wetting water (blue, A* = 14.5) and rapeseed oil (red, A* = 5.8) droplets with robust composite interfaces on a mesh with D* = 2.2. Right inset image shows water (A * = 2.1) in a robust composite interface and rapeseed oil, with a lowered robustness factor (A * = 0.9), wetting a textured surface with a higher spacing ratio D* = 5.1. SEM micrographs of the three meshes are shown on the right. [5]. 2.6 Impact of openness of the weave on breakthrough pressure, Pb A surface can display high water repellency, and therefore high apparent contact angles, without necessarily forming a highly robust interface. As with wettability, openness of weave and its dimensionless parameter, D*, also affect robustness, A *, and the breakthrough pressure, Pb. In this study, then, feather structure is primarily characterized by the effective D* approach, 31 which is presented as more adequate than microscopy for studying feather-water interactions. Surface texture was quantified in terms of an effective spacing ratio (D* -- Deff), and robustness factor values for the various species were generated. The D* and A* parameters connect the structure of bird feathers to ecological behavior, specifically related to bird interactions with water. In a laboratory setting, the connection between D* and A * and diving depth can be practically demonstrated by submerging a wire, woven mesh into water, which is analogous to the submersion of bird feathers during water contact (Figure 11). As a mesh is submerged increasingly deeply in liquid, the pressure difference across the liquid-air interface (P) increases, and the solid-liquid-air composite interface increasingly sags and presses against the air pockets (Figure 1 lb-d). Once it reaches the breakthrough pressure, (P = Pb), the composite interface is driven to a fully-wetted state. Trapped air is displaced by penetrating water, as seen in the bottom segment of the meshes in Figure 11 a, which contrasts two meshes, one with low and one with high D*. The former has a high Pb and thus maintains the robust, composite interface against higher pressures at lower depths: the latter can only withstand low pressures (low Pb), and thus, submersion below a shallow level of water easily fully wets the mesh. The same can occur to bird feathers as birds dive. and remain in water for extended periods of time, which may also dictate the time birds will spend submerged and on land in wing-spreading posture to dry. As explained in the previous section, surface tension is directly varied instead of pressure to generate the D* and A* values required to characterize feather topography and to compare to diving depth trends. Thus, the mathematical framework applied to meshes, fabrics, and micro-fabricated surfaces will be applied to these species, yielding effective D* values using known variables 32 * including contact angle measurements, not visual inference as done in past research efforts. A and Pb values from these feathers allow for the estimation of a maximum diving depth, which at least ranks the species in relative order of diving depth practiced when it not also provides quantitative values comparable to those observed. (b) (a) Pressure (Pa) rO "100 (c) n200 *300 . 400 (d) . 500 low D - high Pb high D -> low Pb Figure 11. Model of bird feather submerged in water. Evolution of the solid-liquid-air composite interface formed between a wire, woven mesh and the water in which it is submerged. (a) A snapshot of two woven meshes with same R submerged in a cylindrical tank of de-ionized water: the left with D* = 2.2 and the right with D* = 5.1. Pressure is indicated in Pascals (Pa) for the corresponding depths of the tank, and every 100 Pa corresponds approximately to 1 cm in height of the water column. (b) - (d) Progression of the solid-liquid-air composite interface corresponding to the same woven mesh under increasing pressure differential values of 50, 100, and 300 Pa, respectively, as simulated using Surface Evolver@ FEM software. Blue, light blue, and red color represent wet solid, liquid on air, and dry wire surface, respectively. Inset in (b) shows the structure of the dry wire surface. Adapted from [5]. 33 3 3.1 Experimental Procedures Bird Feather Specimens Through collaboration with world-renowned biomimeticist and expert zoologist Prof. Andrew Parker of the Natural History Museum, London and Oxford University, UK, access to study the set of carefully-selected feather specimens was obtained. Feather samples from twelve birds from six species were furnished by the Natural History Museum (NHM), London, UK and are cataloged in Table 2. No birds were sacrificed specifically for this study. Table 2. A catalog of the twelve bird feather specimen supplied by Dr. Andrew Parker (BMNH, London), containing scientific names, common names, and identification codes. The wing feather of each listed specimen was studi Number Common name Scientific name 1 Reed cormorant Phalacrocoraxafricanus BMNH 1900.1.20.84 2 Great cormorant Phalacrocoraxcarbo BMNH 1895.6.20.121 3 Great cormorant Phalacrocoraxcarbo BMNH 1894.6.20.137 4 European shag Phalacrocoraxaristotelis BMNH 1897.4.19.2 5 European shag Phalacrocoraxaristotelis BMNH 1941.5.30.3280 6 European shag Phalacrocoraxaristotelis BMNH 2007.64.c 7 African darter Anhinga rufa BMNH 1904.11.19.57 8 African darter Anhinga rufa BMNH 1955.6.N.17.5 9 Common shelduck Tadorna tadorna BMNH 1955.3.10 10 Common shelduck Tadorna tadorna BMNH 1992.9 11 Mallard Anas platyrhynchos BMNH 1980.14.43 12 Mallard Anas platyrhynchos BMNH 1980.14.61 Identification code For this project, wing, breast, and belly feathers of these birds were available, and the wing feathers were selected for characterization. Figure 12 displays photographs of the wing feathers studied and of the breast and belly feathers, as well, that are representative of the actual plumage of other individual birds of the same species. 34 (a) Reed or long-tailed cormorant (Phalacrocoraxafricanus) (b) Great cormorant (Phalacrocoraxcarbo) (c) European shag (Phalacrocoraxaristotelis) (d) African darter (Anhinga rufa) (c) Common shelduck (Tadorna tadorna) (b) Mallard / wild duck (Anas platyrhynchos) Figure 12. Photographs of wing, breast, and belly feathers for the six bird species. (a) reed cormorant, (b) great cormorant, (c) European shag, (d) African darter, (e) common shelduck, and (f) mallard. 3.2 3.2.1 Coating Methodology Fluorodecyl POSS (polyhedraloligomeric silsesquioxane) and Tecnoflon@ solution Fluorodecyl POSS (polyhedral oligomeric silsesquioxane) molecules consist of silsesquioxane cages surrounded by eight IH,lH,2H,2H-heptadecafluorodecyl groups [27]. Due to the high density of perfluorinated carbon atoms present in the eight alkyl chains surrounding the silsesquioxane cages, a smooth fluorodecyl POSS surface has one of the lowest solid-surface energy values reported to date (ys, ~ 10 mN/m) [30]. To confer thin, uniform, flexible, and 35 conformal coatings of fluorodecyl POSS to textures of interest, the commercially available Tecnoflon@ fluoroelastomer (BR 9151, Solvay Solexis) (ysv ~ 18 mN/Nm) was employed as the continuous polymeric matrix, and common solvent Asahiklin AK225 (Asahi Glass Company) was used as the solvent for the Tecnoflon polymer and fluorodecyl POSS. Figure 13 shows the chemical structures of fluoro-POSS molecules and the Tecnoflon polymer. (a) Rf\ 0 Rf S' Rf.s S\O'S' /\ - RO 0 R Si (b) 0 . H2 / F2 c-c- CF F2 F2 F2 c- I F---- SiH-2 SR f Figure 13. Chemical structure of fluorodecyl POSS and Tecnoflon@ fluoroelastomer. (a) General chemical structure of fluoro-POSS molecules, containing alkyl chains (Rf) according to the general molecular formula: Rf = -CH 2-CH 2- (CF2)n-CF 3, where n = 0, 3, 5, or 7. For fluorodecyl POSS, n = 7, and thus Rf = -CH 2-CH 2-(CF 2)7-CF 3 (ysv ~10 mN/m). (b) Molecular structure of Tecnoflon (ys,~ 18 mN/m). Fluorodecyl POSS and Tecnoflon are dissolved in common solvent Asahiklin to form the solution used for dip-coating the feathers. 3.2.2 Coating smooth and textured surfaces Silicon wafer substrates were spin-coated with a POSS-Tecnoflon solution (50%-50% by weight, total solids 10 mg/mL) (ys, = 10.7 mN/m) at a rotation speed of 900 rpm for 30 s. The bird feathers were dip-coated in the POSS (50%)-Tecnoflon (50%) solution (10 mg/mL). After immersion for 5 min, the samples were removed and dried in air to ensure complete evaporation of the Asahiklin solvent. 36 3.3 3.3.1 Surface Characterization Contact angle measurements Contact angle measurements and sliding angle measurements were obtained with a ramd- hart 590-Fl goniometer. Advancing and receding contact angles were measured using -5 [tL droplets of various liquids (purchased from Aldrich and used as received). The advancing contact angle represents the wetting of liquid droplets on previously dry surfaces. The receding angle represents the interaction of the liquid with the surface after liquid has already come into contact with the surface. The advancing angle has been the angle of interest for computing effective D* in previous work [26]. Given the main motivation of this work-to study how a bird begins to interact with water during its diving activities-the advancing angle is the more relevant quantity of the two and thus was selected for D* calculations over the receding angle. Both advancing and receding contact angles were recorded and are both are reported for completeness in Table 3 (Section 4.1). Initially, a distinction was drawn between "inner" and "outer" contact angles, or contact angles measured on the inner region of a feather, near the main shaft, and those measured near the edge of a feather's barbs, respectively. Despite knowledge of a regular, highly waterrepellent central region and irregular, wettable distal region documented for the body feathers of great cormorants [7], the inner-outer construct was abandoned for this study after preliminary experiments revealed no statistically significant differences between inner and outer contact angles across species. 3.3.2 Scanning electron microscopy (SEM) Scanning electron microscopy (SEM) was conducted for wing feathers from two of the species using a JSM-JEOL 6060 microscope (Institute for Soldier Nanotechnologies at MIT) at 37 an accelerating voltage of 5 kV. Samples were cleaned in Asahiklin solvent and gold-coated with a thickness of- 5nm in preparation for microscopy. Micrographs were obtained capturing each level of structure on the surface of the feathers. The feature length scale, Rgeometry, was visually measured from the micrographs and taken the as input for the simple 1 -D model of parallel cylinders. Apart from those values used for this purpose, values for Rgeometry were not further considered in this thesis, and dimensionless D* is independent of length scale, regardless. 4 4.1 Results Apparent contact angles for bird feathers from each species Each feather was probed by various liquids, and the apparent advancing and receding contact angles measured (0*adv and 0 rec, respectively) are reported in Table 3. The probing liquids are listed in order of decreasing surface tension: water (yIv = 72.1 mN/m), diiodomethane (ylv = 50.8 mN/m), ethylene glycol (ylv = 47.7 mN/m), dimethyl sulfoxide (yIv= 44 mN/m), rapeseed oil (yIv = 35.5 mN/m), hexadecane (ylv = 27.5 mN/m), and dodecane (yiv = 23.8 mN/m). The equilibrium contact angles (OE) on a flat, fluorodecyl-POSS-coated surface are also provided for comparison as a reference point. Water droplets displayed the highest apparent contact angles, and as surface tension decreased with each of the other liquids, corresponding 0* decreased. The behavior is exemplified by what happens to a feather in water versus what happens to it in oil: water, with higher surface tension, will not wet the droplet as easily as oil, which has the lower surface tension. Equilibrium contact angles and apparent contact angles (*adv or 0*rec) inherently do not provide any information on surface structure; information about structure is obtained using D* [26]. 38 Table 3. Average, apparent advancing and receding contact-angle data (O*adv and 0*rec, respectively) for wing feathers of each species and for chemically identical, flat, fluorodecyl POSS surface (OE) with various liquids. For each species, liquids are listed in order of decreasing surface tension. Bird species Reed cormorant Liquid Water Diiodomethane Ethylene glycol Dimethyl sulfoxide Rapeseed oil European shag (0) 107 5 90 96 5 101 89 1 13 78 0 Water Diiodomethane Ethylene glycol Dimethyl sulfoxide Rapeseed oil Hexadecane Dodecane 142 124 134 120 : 108 0 0 4 2 5 5 10 133 109 120 104 + 90+ Water Diiodomethane Rapeseed oil Hexadecane Dodecane 140 137 137 112 114 0 0 6 7 4 8 9 Water 137 Ethylene glycol Ethylene glycol Dimethyl sulfoxide African darter O*rec 85 0 Hexadecane Dodecane Great cormorant 0*ad& (0) 125 6 103 4 121 + 9 115 3 113 + 8 7 15 12 5 6 6 5 1 0 0 130 115 120 114 100 0 6 5 6 6 6 0 + 2 126 2 3 2 104+ 6 103 8 6 3 7 132 + 3 115 4 107 + 7 Dimethyl sulfoxide 120 112 Mallard Water Ethylene glycol Dimethyl sulfoxide 141 133 125 Common shelduck Water Ethylene glycol Dimethyl sulfoxide 149 10 140 + 5 142 124 6 131 122 124 116 5 *Flat fluorodecyl POSS surface Water Diiodomethane Ethylene glycol Dimethyl sulfoxide Rapeseed oil Hexadecane Dodecane 6 100 2 2 111 98 88 80 75 2 2 3 1 1 9 79 87 80 66 61 60 * Equilibrium contact angles (6 adv and 0rec) for same liquids on a flat fluorodecyl POSS surface provided for comparison 39 4.2 "Effective microscopy" of feathers with D* The 6*adv values along with the equilibrium contact angles (OE) measured on a flat, chemically identical, fluorodecyl POSS surface (Table 3) served primarily as input data in the modified Cassie-Baxter (CB) relation (Equation 1, Section 2.4.1), which yielded effective D* values for the wing feather of each bird species shown in Figure 14 and Table 4. Figure 14 presents the generalized non-wetting diagrams for all six bird species. The effective spacing ratio, D*, for the wing feather of each species was first found by regression of the CB relation to the advancing, apparent contact angle data for each species. Once D* was obtained, it served as the fitting parameter to plot the CB relation as a fit (solid line) for the % contact angle data (large data points in Figure 14). The dashed lines represent the 95 confidence intervals for the D* fit. Good agreement of contact angle data with its CB fit indicates the 1 -D model can adequately characterize the textured surface. The MATLAB@ code used for the fitting procedure, courtesy of Shreerang Chhatre, is attached in Appendix A (Section 9.1). All these feathers form composite interfaces for an array of liquids with a range of surface tensions. By contrast, a natural, uncoated feather lacks the low-surface-energy coating of fluorodecyl POSS and is only aided by its re-entrant surface topography: its barbs and barbules. To verify the expected behavior of these same probing liquids on a natural feather, the apparent contact angles of the probing liquids on uncoated feathers from two of the species, the one with lowest and the one with highest, effective D*, were also measured (Table 5). Those tests confirmed the same monotonic trend of decreasing contact angles with decreasing surface tension of the probing liquids. The African darter had the lower effective spacing ratio (D* = 1.23 0.21) and, therefore, also had lower contact angles but resisted wetting by hexadecane (yiv = 27.5 mN/m). By contrast, the common shelduck had the highest D*, 40 (a) 180 90 120 180 1.0[ 1X20 0 60 90 0 0.5- 60 , 0.5- (b) 0 -0 60 1.0 60 90 0.0 (0) (c) 12 -0.5 0.0 cos9 0.5 1.0 120 90 60 0 wV 0 -0.5 0 -1.0 -1.0 -0.5 180 120 D =1.36 (d) 0.0 cos9. |180 0.5 1 .0 60 0 0 C) 90 0 1.0 F 0 120 0.39 / / 180 1.0 0.21 - -1.0 -1.0 = 1.23 - -0.5 - 0.5 - 60 0.0 p -1 D -1.0 .0 -0.5 0.0 cos9 120 90 = 1 61 0 25 -0.5 1120 1.018 0 0.5 -0.5 120 0 180 1.0- 60 0.5 (I) 0 60 0 -1.0 (*) 180 .0- d0 .) 7.. ';.d0 -0.5 (e) 90 0.0 900 0 60 0.0 '9 1, 77 1 0.34 -120 0.5 1.0 60 0 80 (0) 90 0 A 7 0 .- 0.0 90 1-/ 0 d.dO 60 90 ' 0 20I -0.5 - -1.0 -1.0 D =1.78 -0.5 0.0 0.5 -0.5 - 120 0.18 ' 180 10 -1. -1.0 1.0 ' =2.28 0.22 1 180 -0.5 0.0 0.5 1.0 cosO. cosO Figure 14. Generalized non-wetting diagram for dip-coated bird feathers from each species. Cosine of the apparent, advancing contact angle (6 adv) is plotted against the cosine of the advancing contact angle on a smooth silicon wafers (OE) spin-coated with the same solution as the dip-coated feathers. The effective spacing ratio, D*, for each specie's wing feather is first found by regression of the Cassie-Baxter relation to the contact angle data for each species (Equation 1, Section 2.4.1). The solid lines correspond to the Cassie-Baxter (CB) equation plotted for each D* value inset in the diagram; the dashed lines indicate the 95 % confidence intervals for the respective CB plots. The data plotted on each non-wetting diagram corresponds to the wing feathers tested for each bird species, in order of increasing effective D*: (a) African darter, (b) reed cormorant, (c) great cormorant, (d) mallard, (e) European shag, and (f) common shelduck. Inset in (b) shows a typical water droplet on a wing feather of reed cormorant. 41 Table 4. Effective spacing ratio, D*, values for each bird species Bird species D*,ff African darter Reed cormorant Great cormorant Mallard 1.23 + 0.21 European shag Common shelduck 1.36 1.61 1.77 1.78 2.28 0.39 0.25 0.34 0.18 0.22 2.28 h 0.22, which agrees with its lower wettability-higher contact angles-but lower robustness, as evidenced by it being fully wetted by hexadecane when the other species of lower D* is not (see Section 2.5 and 2.6 for concept). The data also showcases the effect of the fluorodecyl POSS coating, which increases liquid repellency, and thus 6, and generally renders oleophobic surfaces that would otherwise not repel oils. Table 5. Average, apparent advancing and receding contact-angle data (O*adv and 6*rec, respectively) with various liquids for wing feathers of the species with highest and lowest D*, African darter and common shelduck, respectively. For each species, liquids are listed in order of decreasing surface tension. Bird species African darter Common shelduck Liquid Water 0*adv (0) 128 2 Ethylene glycol 104 5 0*rec (0) 116+ 5 93 3 Dimethyl sulfoxide 90 + 8 77 + 7 Hexadecane 45 3 31 3 134 1 125 + 3 129 109 2 4 Water Ethylene glycol Dimethyl sulfoxide Hexadecane 114 7 0 86 5 0 As an additional, dynamic test for wettability, sliding angle measurements were taken for a few of the liquids on selected feathers. In those experiments, the contact angle becomes a sliding angle: the incipient angle at which the droplet begins to slide off the surface as the surface is gradually tilted. Sliding angle data is included as Appendix B (Section 9.2). 42 4.3 Scanning electron microscopy (SEM) of selected feathers Past studies have relied on photographic and microscopic techniques in attempts to * characterize feather structure and wettability [6,8-10,22], but this study presents the D* and A frameworks as a more adequate alternative for the characterization of complex surface topography. Scanning electron micrographs obtained for the African darter (D* = 1.23 and common shelduck (D* = 2.28 0.21) 0.22) showcase the complexity and diversity of feather topography (Figure 15). The micrographs start with the main shaft and barb structure of the feather and, with increasing magnification, reveal underlying structure in a hierarchical manner: - from barbs (Figure 15a, i), to barbules (c, d; k, 1), to smaller scale rods on the African darter (e h) and branch-like structures on the common shelduck (m - p) on the order of 10 pm. Additionally, the coverage of different areas on the feather surfaces highlights other differences in structure between the more central areas and distal areas (edges) of the feathers. Micrographs (c) and (k) can be contrasted with (g) and (o), for instance, to show differences in the order and density of barbules between central areas and distal areas; (d) and (1) and (h) and (p) can likewise be contrasted as barbule tips on the feather edges. Finally, the difference in D* between the two bird species is noticeable. The African darter apparently has a tighter weave, and hence a lower D*eff, whereas the common shelduck has a more open weave and higher D*eff. Also apparent in such ramage is the difficulty of manually measuring R and D to find D* values, which regardless of accuracy nevertheless fails to fully assess how topography contributes to wettability. Indeed, if manually measured at all, R and D should be measured with respect to barbules, because barbules contribute most to the surface area of the feather with which a liquid would come into contact. Unfortunately, barbules are also finer, can pack differently from barbs, and can possess even finer features, as in the case of the 43 barbules of the common shelduck, which branch out with even smaller-scale structures that resemble the split ends in human hair (Figure 15m, n). This complexity, however, is not unconquerable, and topography and wettability can still be approached by generating D*eff values and combining them with "Rgeometry" values, taken from the barbules of these micrographs, as approximations of the effective length scale, Reff. With this Reff, D* values, and the contact angle measurements, the robustness of the surface against wetting (A *) and the breakthrough pressure (Pb) can be readily computed. 44 Common shelduck African darter (a) (i) (b)l (j) (c) (k) (d) (1) 45 Common shelduck A frican darter (e (n) (m) (o) (p) 46 Figure 15. Scanning electron micrographs for topography of wing feathers of African darter and common shelduck. Left column (a - h) corresponds to African darter, and right (i - p) to common shelduck. Top-to-bottom order corresponds with increasing magnification for (a - f) and (i - n) of central feather area and for (g, h) and (o, p) of distal (edge) feather area. Colored boxes indicate dimensions of representative areas that were successively magnified and match with micrographs of the magnified areas. Arrow indicates when successivley magnified area is beyond visible area. Accelerating voltage, 5 kV. Spot size of 50 at a working distance of 8 to 12 mm. 4.4 Critical diving depth from robustness parameter, A*, and breakthrough pressure, Pb Robustness parameter, A *, and breakthrough pressure, Pb, can be obtained for each species from Equation 2 (Section 2.4.2). Instead of finding a feature length scale, R, for each species, Reff is approximated by the Rgeometry values measured from scanning electron micrographs like those in Figure 15. Both Rgeometry and Dgeometry were measured for the barbs and barbules of the African darter and common shelduck. From these values, D *geometry was computed, and all three quantities are reported in Table 6. African darter had a D*eff Of 1.23 0.21 and the common shelduck a D*effof 2.28 0.22, both of which are approximately the same as the D*geometry values computed from the R and D of barbules, but far from those computed from the R and D of barbs. Agreement of D*eff (Table 4) with D*geometry computed from barbule measurements and disagreement with the same quantity computed from barb measurements supports the expectation that barbules will play a greater role than barbs (and rachis) in liquid-feather interactions due to their contribution to surface area. Therefore, calculations for A* and Pb proceeded with Reff taken as the overall average of Rgeometry values based on barbule measurements on the two birds (Rgeometry = 5.4 1 pim). Table 6. Rgeometry and Dgeometry as measured from scanning electron micrographs and corresponding, calculated D*geometry for barbs and barbules of African darter and common shelduck feathers Bird species Structural feature African darter Barbules Common shelduck Rgeometry (pm) Deometry (pm) D *eometry 6.5 0.6 2 0.8 1.4 0.1 Barbs 37.6 2.4 109.9 5.3 3.9 0.2 Barbules Barbs 4.3 11.1 0.4 0.6 5.4 134.9 1.0 3.4 2.3 0.2 13.2 0.9 47 Using the 1 -D model, the robustness factor, A *, and breakthrough pressure, Pb, were estimated for each species with water as the contacting liquid. Based on estimated Pb values, the equivalent swimming velocity, v, due to dynamic pressure (P= pv2 , where p is fluid density) and the expected "breakthrough depth" were also calculated. The breakthrough depth, hb, represents the critical depth at which the breakthrough pressure of the feather structure is met by the bird in the course of diving. As expected from the A* equation (Equation 2), for a fixed liquid and feature length scale, R, a higher D* is accompanied by a lower robustness factor and lower breakthrough pressure. These variables, collected in Table 7, are those quantities that may help connect feather structure to bird behavior. Table 7. Compilation of fitted D*, calculated A*, expected Pb, equivalent breakthrough swimming velocity, and breakthrough depth for each bird species. African darter Effective spacig ratio, D ,f(kPa) 1.23 0.21 Reed cormorant Great cormorant 1.36 1.61 Mallard European shag Common shelduck Bird species 5 Robustness factor, A* Breakthrough Pressure, Pb Equivalent1 2velocity, v = (2PP) / (M s_') 13.6 Breakthrough depth, hb (M) 9.4 1727 92 0.39 0.25 1033 544 55 29 10.5 7.6 5.6 3.0 1.77 1.78 0.34 0.18 403 396 22 21 6.6 6.5 2.2 2.2 2.28 0.22 201 11 4.6 1.1 Discussion 5.1 Apparent contact angles used to assess wettability of bird feathers From Table 5 of O*adv and O*rec values for uncoated feathers and Table 3 for coated feathers, contact angles are statistically different. Feathers previously wetted by low surface tension liquids become liquid-repellent after coating. The wing feathers of the common shelduck, mallard, and great cormorant achieved the highest apparent contact angles with water (O*adv in the range of 140 - 150'), whereas they saw lower O*adv for dimethyl sulfoxide droplets 48 (- 120 - 130'); the great cormorant, which was also probed by rapeseed oil, had even lower contact angles for that liquid, with 6*adv = 108 100. With a lower-surface-tension liquid, hexadecane, the reed cormorant feather resists wetting and supports the droplet at a composite interface, whereas the feathers of great cormorant and European shag do not and are fully wetted by the liquid. Ideally, the contact angle data can be refined into one parameter that characterizes texture, namely D*. 5.2 Effective microscopy (1-D model) characterizes feather texture The "effective microscopy" by which the structural details of each feather were captured in a single parameter yielded D* values for the wing feather of each species (Table 4). This effective D* analysis employed throughout the present work managed to characterize the complex, delicate feather surfaces about as well as it has highly ordered, mesh surfaces in past work [28]. For comparison, the average error in D* for all bird feathers was ~ 0.27, close to the 0.2 reported for dip-coated, woven meshes and smaller than the 0.6 error recorded for dip-coated, carbon paper composed of randomly oriented, cylindrically textured microfibers [28]. To date, then, including in this study, a 1 -D model of D*eff has proven sufficient for characterizing diverse surface textures, and need for a higher-dimensional model for contrast against the 1 -D model has not led to any new understanding of feather wettability (see Section 7). The species whose contact angle data had the greatest disagreement with the Cassie-Baxter (CB) fit, the reed cormorant, is difficult to address because of the small data set: it is the only species for which only one feather was tested due to availability. 49 5.3 Effective D* is better indicator of feather wettability than D* from geometry In evaluating our effective microscopy technique as applied to bird feathers, a comparison with literature values of D* = (R + D) / R obtained geometrically (henceforth, "D *geometry), namely those reported by Rijke and in related work, is useful. Effective D* values (D*eff) as obtained from the CB fits (Figure 14) are reproduced alongside D*geometry values obtained by visual measurements in Table 8. In this regard, literature unfortunately provides limited information on D* values, which are only cataloged for some species and for varying feather categories. Table 8 indicates when wing feather values for the particular species were not available and another value was used and when other values were also included based on relevance. Table 8. Effective D* values (D*eff) from this work and D* et = (R + D) / R values from the literature Bird species African darter Reed cormorant Great cormorant Mallard European shag Common shelduck * from breast feathers in D*eff 1.23 +0.21 (R + D) /R Reference ~ 2-3, 4.5 [10,11,22] 1.36 0.39 1.61 + 0.25 3-4,4.3* 4.8* [10] [6] 1.77 + 0.34 1.78 0.18 ~ 6-7, 5.9* [10] -- -- ~ 9-0, [10] 2.28 0.22 Rijke's study [6] As D*eff increases, the (R + D) / R values increase, as well. A comparison between calculated D *eff values and the D *geometry values shows they both follow the same general order; however, there is no quantitative agreement. Admittedly, a few of the only available previous measurements in the literature were on breast feathers, not on wing feathers, so some of the mismatch is not surprising, but lack of literature data aside, the D* values do follow a general trend. 50 The objective of this work was to study liquid-feather interactions, which is the reason for characterizing feather texture. D*eff is a macroscopic measurement employing a wide range of liquids over many parts of a feather. Depending on surface tension, different liquids wet the same textured surface to different extents. This sampling better mimics water-feather interactions under different hydrostatic pressures. For example, probing a feather with a rapeseed oil droplet compared to a water droplet is roughly equivalent to subjecting it to twice the pressure under water (see Section 2.6). By using a set of contact angles measured for different liquids, the effect of hydrostatic pressure on wettability is captured, as the technique is based on actual liquid-feather interactions, just like the water-related behavior of birds. On the contrary, D* measurements based on microscopy (D *geometry) explore only a small portion of a surface a time. Moreover, D*geometry characterizes the physical appearance in a top view, and not the wetting behavior. The re-entrant portion of the texture, (i.e. the portion below the top half of the structural features) is not visualized, and therefore is missed, by microscopic or visual techniques. The scanning electron micrographs displayed in Figure 15 attest to the complexity and diversity of feather structure and even show signs of hierarchical structure, qualities not easily characterized in a quantitative sense by data obtained from visual methods. Finally, bird feathers are fragile, and electron microscopy is relatively slow and expensive. An effective D* is directly relevant to wettability and captures structural details otherwise missed. 5.4 Role of wettability (O*adv) and feather texture (D*) on ecological behavior As the Cassie-Baxter (CB) relation shows (Equation 1), everything else being the same, an increase in D* means an increase in 6*, and thus, improved non-wetting. For example, the common shelduck displayed the highest 9* and had the highest D*, as shown in Table 3 and Table 4, respectively. Evident directly from the CB relation for a 1 -D cylindrical array, this 51 trend also generally held for the feathers of the six species with distinct D* and is graphically conveyed in Figure 14. Those graphs emphasize that a simple comparison of contact angle data is not the objective, but rather, the connection between topography and wettability behavior that D* helps explore. A revisited version of Table 1, Table 9 presents the values in this study that were initially posited and now confirmed as relevant for analyzing bird-water interactions. Table 9. Catalog for the six species of metrics of interest for this study for analyzing bird-water interactions. Species are arranged in order of increasing D*, decreasing A*, and decreasing, predicted hb. B Bird species African darter Diving depth Diving speed (m s-1) <5 0.19 Wingspreading 0.161 D*ef Robustness factor, A* Y 1.23 0.21 1727 Breakthrough depth, hb (M) 9.4 Reed cormorant 5 - 6* 0.7-0.85 Y 1.36 0.39 1033 5.6 Great cormorant 4.7, < 101 1.1-2.1 Y 1.61 0.25 544 3.0 403 2.2 N 1.77 0.34 Dabbling Mallard Dabbling 396 2.2 Y 1.78 0.18 1.7-1.9 33 -35 European shag 201 1.1 N 2.28 0.22 Dabbling Common shelduck Dabbling * from neutral buoyancy experiments, not natural observation; T horizontal traveling speed, not diving speed; I usually < 10 m, but can dive to depths of 35 m [17]; Wing-spreading: Y = predictably, N = never; Dabbling = dabbling species, not primarily divers The D* framework as applied to birds reveals a correlation between feather structure and diving behavior that microscopy and photography alone cannot reveal. In discussing diving trends, it is taken for granted that the birds are not limited by lack of strength to propel themselves to desired depths. Feathers are expected to play a significant role in diving behavior, as diving for birds such as cormorants is believed to be quite an energetic investment relative to other divers due to poor insulation and less-efficient foot propulsion [37]. Effective openness of the weave as captured by D*eff for the birds of Phalacrocoraxspecies (the cormorants and shag) serves as a starting point for discussing their behavior. From reed cormorant, to great cormorant, to European shag, as the diving depths increase, so does D*. As for the ambiguity of which dives less deeply, whether the reed or great cormorant based on actually documented depths, the great 52 cormorant, while recorded in literature as typically diving up to 4.7 m and < 10 m, actually does show more range than the reed cormorant. In fact, the great cormorant is credited as having feather structure that allows for partial plumage wettability, meaning it is wettable, but not completely, a feature thought to enable it to be less positively buoyant [7,38]. In light of such studies and observation, then, it is likely great cormorants routinely dive deeper than reed cormorants, and their differences in D* predict the same. Remarkably, the highest D* value among the cormorants and shag studied belongs to the European shag, which without doubt exhibits the deepest diving. Shags are also estimated to have lower energetic costs to dive than great cormorants, which is attributed to the streamlined body shape-hinting at the idea that individual feather structure is not the only factor dictating diving ability-and their thick layer of trapped air [37]. In general, birds in the Phalacrocoracidaeare known for spending extended periods underwater, perhaps contributing to their need to assume spread-wing postures to dry. Closely related to cormorants (Phalacrocoracidae),the African darter displays significantly different behavior that agrees with its greater wettability. The species with the lowest D* of the set (D* = 1.23 0.21), Anhinga rufa is known for its low buoyancy, causing them to look like snakes, with only their neck protruding from underwater as they forage. Unlike the cormorant and shag, the darter and others in the Anhingidae family are considered shallow divers. Similar to its low D* value relative to the other species, the darter's plumage is assigned the label of "fully wettable" by past studies. In addition to that, they possess thinner, less spongy skin, and denser bones, and smaller air sacs [16]. These features, and the fact that its feathers are the most wettable of the species characterized by D* and 0*, make the bird suitable for its shallow foraging, which is characterized as slow and stealthy compared to other diving birds. They would also serve as good reasons to practice wing-spreading in-between dives. 53 The extension of the D* framework for aquatic birds is not limited to divers, however. Even behavior already seen as somewhat self-explanatory, such as the absence of spread-wing postures in the lives of the two dabbling duck species in this set can be revisited and now more directly attributable to feather topography. Dabbling ducks usually feed on the surface, a trait that does not demand much, if any, diving from them. In fact, the liquid-repellency of mallard and shelduck feathers as characterized by 6 *adv and D* reflect just how their feather structure suits their practices: their O* values with water are two of the three highest, well in the range of 1400 to 1500, and D* for their wing feathers are likewise in the top three highest, contributing to a greater fraction liquid/air interface that drives water droplets to easily bead up and roll off a bird's coat. This is important during their surface dipping or upending and a general benefit for such birds that spend much of their time in water. Documented ecological characteristics such as the great cormorant's partially wettable plumage, the African darter's notoriously low buoyancy, and the dappling ducks' high waterrepellency warrant a discussion on the robustness parameter, A*, and how in addition to wettability, robustness at the water-feather interface correlates with behavior. 5.5 Role of robustness factor, A*, and breakthrough pressure, Pb, on ecological behavior For the deep-diving birds, the expected breakthrough depths are significantly smaller than actual diving depths. This simply parametric analysis indicates feathers are wetted below this particular depth, and beyond an equivalent threshold velocity of swimming, both tabulated in Table 7. The reed cormorant and the dabbling birds are the only ones for which the expected breakthrough depth quantitatively agreed with observed behavior. On the other hand, the behavior metrics for the European shag, for example, which appears to have an good balance of both water-repellency and robustness, were captured at least in concept if not quantitatively by 54 its relatively high D* and low A*: after its deep and long dives [15], its feathers will undoubtedly need drying, according to the low breakthrough depth for the shag (hb = 2.2), which is in the low range of the dabbling birds studied. Also, the A* may be representative of more than trends, and may also link specific behavior with structure. For example, the high robustness of the darter, while not following the expected trend with diving depth, may be key for explaining Rijke's observation that although the darter emerges dripping wet from diving, remarkably, it is able to immediately become airborne. The analysis does show agreement in ranking order of diving depths and capabilities, and where there is disagreement in magnitude between breakthrough depths and actual diving depths, it is possibly attributable to other unexplored factors, such as the packing of whole arrays of feathers, or more likely, a limitation of the 1 -D, cylindrical model. As for the former, there may be reason to expect higher-order structures to operate on the same physico-chemical principles as its feather subcomponents, suggestion that explanation may be less probable but also difficult to test. While the A * values obtained by the simple 1 -D, cylinder model do not follow the exact trend expected for diving depths, they do correlate with other water-bird phenomena, namely wing-spreading and length of time spent submerged or on water. The observation that cormorants and shags spend much longer times submerged, whereas the African darter spends much less time does not necessarily agree with its high calculated robustness factor alone, but combined with its D* parameter (see Section 5.4), the calculated parameters together paint the more complete picture that describes its behavior. Unlike deep or shallow diving birds, the mallard and common shelduck don't require feathers that resist water penetration at great depths or against high diving speeds or fast 55 underwater swimming. However, their calculated A * values and breakthrough depths indicate their feathers do adequately equip them for spending extended periods of time on water and are generally robust; the expected Pb is a sufficient threshold for them to carry on their water activities as dabbling ducks unrestrained above a predicted 1 - 2 m of water. Young shelducks, for instance, reportedly speedily dive just below the surface when in danger or approached while the adult attendants take flight. As dabbling birds that do not face great hydrostatic or dynamic pressures, these ducks already don't seem to have any need for the spread-wing postures the other aquatic birds exhibit, and the robustness of their feathers more so suggests they would not need to use such techniques, even if they were to partake in more demanding water activities of their own. Theoretically, like for many textured materials, the longer a bird remains in water, the more vulnerable its feathers may become to impinging water [1]. Also like textured surfaces in general, previously wet feathers that do not fully dry are predicted to be more susceptible to wetting. To guard against these two threats, even dabbling birds appear to require robust feathers, while the other birds combine the wing-spreading, drying technique with whatever robustness their feathers offer. 6 Conclusions It is demonstrated that the 1 -D, effective-D* paradigm can effectively be used to characterize the wettability of complicated textures such as bird feathers. Using the D* analysis in conjunction with the robustness parameter framework allows us to a prioridetermine breakthrough pressures of bird feathers. This allows us to compare the diving depth up to which a bird's feathers will theoretically not wet. Alternatively, it also predicts a bird's resistance to 56 wetting when it comes into contact with water contaminated with oil or other lower-surfacetension liquids. In addition to diving, other water-related behavior of aquatic birds such as wingspreading, shallow foraging, and dabbling were correlated to the calculated metrics, namely the effective spacing ratio, robustness factor, and breakthrough depth, with notable agreement for several of the aquatic bird species. 7 Future work As this project has demonstrated, while D* and A * offer useful insights for the behavior of aquatic birds studied, not all behavior is captured by the model of a 1 -D cylindrical array, and future work is warranted. Complementary wettability experiments with higher-order structures of feathers, such as feather groups, feather coats, or whole birds would help more clearly draw connections between the parameters quantified in this work. Also, a better model or surface evolver simulation should reveal further details of bird-feather wettability, especially given that the breakthrough depths predicted by the 1 -D model are of the same order as the observed diving depths. Other areas of interest for follow-up study encompass various ecological and biological variables. Molting intensity, which can affect the integrity of feathers and their structure, for example, would be one variable to track for a set of species. Similar to molting, seasonal variations might be studied to reveal possible differences in feather structure throughout a year. Feather diversity for a single bird based on different locations on its body could also come into play, and studying breast and belly feathers in addition to wing feathers, for example could contribute to the picture of bird/water interactions, as they may show distinct effective spacing ratios, such as significantly different D* for feathers from relatively fluffier areas of the body. 57 As more information on these birds enters the body of literature, there will be more ways to frame bird behavior questions. 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Rijke A, Jesser W. The wilson journal of ornithology 2010;122:563. Elowson AM. The auk 1984;101:371-383. Rijke AM. The auk 1987;104:140-142. Mahoney SA. The auk 1984;101:181-185. Hustler K. Ibis 1992;134:229-236. Ropert-Coudert Y, Gremillet D, Kato A. Mar biol 2006;149:415-422. Wanless S, Burger AE, Harris MP. Ibis 2008;133:37-42. Ryan PG. J avian biology 2007;38:507-514. Hatch JJ, Brown KM, Hogan GG, Morris RD. Great cormorant (phalacrocorax carbo). In: Poole A, Gill F, editors. The Birds of North America. Philadelphia, PA.: The Birds of North America, Inc, 2000. Frederick R. Troeh, Roy L. Donahue, editors. Dictionary of agricultural and environmental science 2003:112. Wilson RP, Wilson M-PT. Journal of animal ecology 1988;57:943-955. Birdlife international (2011) species factsheet: phalacrocorax africanus 2011. Svensson L, Mullarney K, Zetterstrom D, Grant PJ. Collins Bird Guide. London: HarperCollins Publishers Ltd, 1999. Rijke AM, Jesser WA, Mahoney SA. 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Proceedings of the national academy of sciences 2008;105:18200 -18205. [31] Wenzel RN. Industrial & engineering chemistry 1936;28:988-994. [32] Cassie A, Baxter S. Transations of the faraday society 1944;40:546-551. [33] Quere D, Callies M. Soft matter 2005;1:55-61. [34] Marmur A. Langmuir 2003;19:8343-8348. [35] Nosonovsky M. Langmuir 2007;23:3157-3161. [36] Michielsen S, Lee HJ. Langmuir 2007;23:6004-60 10. [37] Enstipp MR, Grdmillet D, Lorentsen S-H. J. exp. biol 2005;208:3451-3461. [38] Wilson RP, Hustler K, Ryan PG, Burger AE, Noldeke EC. The american naturalist 1992;140:179-200. 60 9 Appendices Appendix A: MATLAB® code for D* fitting procedure 9.1 function estimateDstarfeathers This function reads the contact angle data from an excel spreadsheet and fits the cylindrical cassie baxter equation to estimate D written by Shreerang Chhatre global costheta; from ACU fa-bric- Dstar calculation.xls read the- data first for Bird Feathers v2.xls', xlsread. 'Graphs contactangle : 'graph', 'C5:Cll'. ; 'graph', 'U5:Ull'. ; cos. contact-angle pi.180.; cos theta : contact angle xlsread. 'Graphs costhetastar cos.contact_angle pi.180.; for Bird Feathers initial - Vstar0 ' 2; v2.xls', guess actual function . Dstar,r,J,SIGMA,mse. : fun,Dstar0. nlinfit. costheta,costhetastar, Dstar mse ci : nlparci. Dstar,r,'covar',SIGMA.; ci. 1... 2 .ci. 2. delta : Now plotting- the datapoints along with the fitted CB relation 100; n xx : 1,1,n: ; linspace. figure; plot. xx-,- fun. Dstar,xx , plot. costheta,costhetastar, fun. Dstar delta,xx, xx, fun. Dstar delta,xx.. ; hold on; 1 1. . ; 1 1 axis. . xx, 'o'. ; hold on; from regression', 'D^ legend. 'D delta'. ; xlabel. delta', 'D cos\thetaE'. ; ylabel. 'cos\theta' hold off; return; function y : 1 y fun. D,x. .1. D. . sin. acos. x. . pi acos. x. .. x.; the function return; Figure 16. MATLAB@ code used to estimate the dimensionless spacing ratio, D*, for each bird species. Takes contact angle data as input to fit the Cassie-Baxter equation for parallel cylinders and output D* as the fitting parameter. Courtesy of Shreerang Chhatre. 61 9.2 Appendix B: Sliding angles measured for selected birds and liquids Sliding angles were measured using 15 pL droplets deposited on the feather surface such that upon tilting the droplets slid parallel to the main shaft (rachis) of the feather and perpendicular to the barbs. A tilt angle range of 0 to 900 was spanned. Due to the inconsistency of measurements obtained from preliminary sliding-angle experiments and the ambiguous meaning of the dynamic sliding angle as related to bird-water interactions, the sliding angle experiments were not conducted more extensively, more static contact angle measurements were conducted instead. Table 10. Sliding angle (co) measurements for water and ethylene glycol on wing feathers from species in the Phalacrocoracidaefamily. Liquid Bird species (Owarer (0) Cwethylene glycol (0) Reed cormorant 90* Great cormorant <5 30 7 27 5 40 20 European shag * did not slide at 900, but exhausted tiling range without sliding off. 62