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31295000586429.Pdf (5.941Mb) THE SYSTEMATIC STATUS OF FUNDULUS KANSAE AND FUNDULUS ZEBRINUS by CHARLES THOMAS EVERETT, B.S. in Ed. A THESIS IN ZOOLOGY Submitted to the Graduate Faculty of Texas Tech University in Partial Fulfillment of the Requirements for the Degree of MASTER OF SCIENCE Approved August, 1972 8QJ> f^tf^/oi"^ m Jin ACKNOWLEDGEMENTS I am deeply grateful to my major advisor, Dr. John S. Mecham, for his direction of this thesis. I am thankful to Drs. Francis L. Rose and Robert W. Mitchell for the time and equipment they provided. I sincerely thank Mr. Mike Bishop, Mr. Scott Simpson, Mr. Greg Mengden, Mr. Tony Mollhagen, and especially my wife, Donna, for their assis­ tance in collecting the fish of this study. I am grateful to Dr. Joe R. Goodin and, again. Dr. Francis L. Rose for their constructive criticism of this manuscript. This research was supported in part by National Science Founda­ tion Grant {GB-13791) under the direction of Dr. John S. Mecham. 11 CONTENTS ACKNOWLEDGEMENTS ii LIST OF TABLES iv LIST OF FIGURES v I. INTRODUCTION 1 II. METHODS AND MATERIALS 3 III. RESULTS 10 Morphological Analysis 10 Electrophoretic Analysis 12 IV. DISCUSSION 32 V. SUriMARY 3 9 LITERATURE CITED 41 111 LIST OF TABLES Table Page 1. Analysis of Variance, Single Classification Males v.s. Females 25 2. Regression of Morphological Characters on Distance from Southernmost Locality 26 3. Descriptive Statistics for Pectoral Rays, Dorsal Rays and Vertical Bar Number-Males 28 4. Descriptive Statistics for Anal Rays and Principal Caudal Rays 30 5. Degree of Correlation between Scale Number and Vertebral Number 31 IV LIST OF FIGURES Figure Page 1. Sample Localities for Fundulus zebrinus 16 2. Typical Male and Female F. zebrinus 18 3. Descriptive Representation of Lateral Scale Number with Respect to Locality 20 4. Variation in Eye Diameter with Respect to Locality 22 5. Electrophoretic Pattern Differences between F. zebrinus and Cyprinodon rubrofluviatills 24 V CHAPTER I INTRODUCTION According to the current literature there are two spe­ cies of the genus Fundulus in the saline, alkaline waters associated with the upper regions of the Central Plains drainage systems. These two fish, F. zebrinus Jordan and Gilbert and F. kansae Garman, exhibit morphological charac­ ters not observed elsewhere in the genus. These include a lengthy convoluted gut and weak, slender pharyngial teeth. Two characters have been utilized in differentiating the two fish. The first character, lateral scale number, is found in many recognized keys (Blair, et al., 1968; Eddy, 1969; Knapp, 1953). In F. kansae the lateral series number is given as 52 to 64 scales while in F. zebrinus the scale count varies from 41 to 49. The other character, eye diam­ eter, has been only infrequently mentioned (C. L. Hubbs, 1926; Koster, 1957). Hubbs simply reported that F^. zebrinus had a larger eye. Koster compared the eye diameter to the width of the preorbital bone. In F^. kansae the width of the preorbital was reported as being two-thirds or more of the diameter of the eye while in F_. zebrinus the bone width is no greater than one-half to two-thirds the diameter of the eye. According to Koster, the preorbital size is essen­ tially the same size in both species. There has been some disagreement as to the ranges of the two fish, but the concensus of most contemporary workers is that F. kansae is found from South Dakota to northern Texas and eastern New Mexico, or more generally the Arkansas River basin east to Missouri; F^. zebrinus is found in the Brazos, Colorado, and Pecos River drainages of Texas and New Mexico. Several authors (Miller, 1955; Metcalf, 1966; Pfliger, 1971) have suggested the possibility of conspecificity be­ tween F. kansae and F. zebrinus but have lacked sufficient evidence to support this contention. This study was under­ taken with the objective of determining the systematic relationships of the two animals. CHAPTER II METHODS AND MATERIALS Specimens for analysis were collected at the 20 local­ ities indicated in Figure 1. These localities were dis­ tributed from the lower portion of the Pecos drainage in southwestern Texas northward to the Smokie Hill River in the west-central portion of Kansas (least distance approxi­ mately 573 miles). Eight distinct drainages were sampled, these being from north to south; Smokie Hill River, Arkansas River, Cimarron River, Canadian River, Red River, Brazos River, Colorado River, and Pecos River. Samples from numer­ ous localities within the Canadian, Red and Brazos River drainages were studied in order that the supposed range limits of the fish could be closely inspected. All fish were collected with a fine mesh, 20 foot seine. The fish were most generally found in the stream bed proper where the water was shallow with a moderate rate of flow, or in the side pools adjoining the main body of water. The fish definitely seemed to prefer a substrate of sand or fine gravel. The fish were kept in styrofoam containers for transpor­ tation to the laboratory. Upon arrival in the laboratory a random sample of the fish were preserved in 10% formalin; the remaining fish were kept in aquaria, styrofoam coolers, and large plastic containers. All fish were kept in an 3 environmental room with the water temperature stabilized at 20.0 - 1.0°C. The fish were maintained on a diet of shrimp flakes and commercial tropical fish food. Freshly preserved fish were allowed to set for several days before measurements and counts were taken. This was done to allow completion of preservation, as these fish would be grouped with aged museum specimens. Setting also affected erosion of the mucous layer covering the scales. This layer seriously hinders the counting of scales. Four measurements were recorded (to the nearest 0.1 mm) for each fish. These were: standard body length, predor- sal length, eye diameter, and body depth immediately poste­ rior to the operculum. A binocular, widefield scope was used as an aid in taking all measurements. Counts were made of six meristic characters for each fish. These in­ cluded numbers of pectoral rays, dorsal rays, anal rays, principal caudal rays, vertical bars, and lateral scales (from the first scale in contact with the operculum to the last large scale in the region of the hypural plate). Scales were counted on both sides and averaged if there was a difference. Any mean value ending in 0.5 was raised to the next whole number. The scales in these fish are normally quite difficult to count, and a technique was devised which greatly simpli­ fied this problem. After all other measurements and counts 5 were taken, the fishes were placed in close proximity (7-10 cm) to an incandescent bulb and allowed to dry slightly. They were then placed in a solution of methylene blue in distilled water (1:200) for a period of approximately three minutes. They were again dried for three to five minutes under the lamp. With the aid of a dissecting scope the scales could then be easily and accurately counted. Upon return to the formalin the fish gradually lost their blue color and were in no way harmed. The ray counts generally proceeded without difficulty, but on occasion drying or backlighting was necessary. The fish were also sexed at the time of measurement. This is a relatively simple procedure. The males have much bolder barring than do the females (Fig. 2). The male has a rounded anal fin while in the female it is angular. The female also has a low sheath around the anterior portion of the anal fin. Another sexual difference, previously un­ recorded, is a black spot located above the pectoral fin just posterior to the operculum. This spot is consistently present in the males and absent in the females. When possible, thirty animals, including fifteen males and fifteen females, were analyzed from each locality. A total of thirteen localities afforded this number of animals. Where only a lesser number could be obtained, the animals were measured and counts taken in the same manner but the 6 sample was not included in the analyses of variance or re­ gression analyses as unequal sample size greatly burdened the analysis. All samples were analyzed in terms of descriptive statistics. Each character was first analyzed for the presence of sexual dimorphism. This was accomplished by single class­ ification, analysis of variance. If sex influenced the phenotypic expression of a character the males and females were compared separately; if sex did not create a bimodal distribution they were combined for study. Data for certain characters suggested clinal variation. To evaluate a possible clinal change in phenotypic expres­ sion the various mean values for each character were re­ gressed against locality on a south-north axis, the southernmost locality being assigned a value of zero. All other values were expressed in miles from this locality. The locality was considered the independent variable and the character, the dependent variable. Twenty animals, five from each of four localities, were examined for possible correlation between vertebral number and lateral scale number. The scale counts were made as previously described except that the scales were not stained, The vertebral counts were made after clearing and staining the fish in a modified method of Taylor (1967) as described by Mitchell (1971). Vertebral counts were taken from the 7 first unfused neural spine to the last neural spine on the hypural plate. Logrithms were utilized where ratios were involved in the evaluation of a character. Instead of dividing one character by another, the log of the smaller value was sub­ tracted from the log of the larger.
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