Abstract The abundance and distribution of microfauna found associated with Prionitis lanceolata at a point chosen to represent an unpolluted, natural environment. The results of this study were compared to the results of a previous study made in the Pt. Pinos area. Also, areas at Pt. Pinos uere resampled to determine the effects of seasonal variation on microfaunal communities. Results indicate that differences in microfaunal community structure observed at Pt. Pinos are greater than those attributable to the effects of natural variation. Seasonal varistion uas found to have little effect in the Pt. Pinos area. From these results a hypothesis for testing for pollution damage in an area uas formulated. 2. Introduction Uhile noticeable differences in numbers of species and diversity indices uere found to exist in the microfaumal communities associated with the three species of algae examined around the polluted portion of Pt. Finos (Clearman, 1970), the role of the effluent in causing these differences may not be clearly stated until more is knoun of the natural variation in com- munities of this type. It is possible that the observed differences fall within the general range of natural variation along the point. In order to provide an ansuer to this question, a study similar to the survey at Pt. inos (Clearman, 1970) uas undertaken at another area chosen to represent an unpolluted, natural environment. This area is an unnamed point, hereafter referred to as Point 42, that lies approximately 1000 meters south of Pt. Pinos. The point is approximately 2/3 the length of Pt. Pinos, but is of a similar conformation, and maintains the general Nu orientation of Pt. Pinos. Uave action at the tuo points is roughly comparable, although Pt. Pinos may recieve slightly heavier wave action at times. By studying algal-microfauna associations on this point, it uas hoped t see hou natural communities compared with those previously observed at Pt. Pinos, thus allowing a firmer statement to be madelofthe effluents effect on the communities studied. Also, key areas, as revealed by the original survey at Pt. Pinos, uere resampled to determine the composition of the communities after a tuo month interval to see hou they compared to the results of the first study carried out during the spring. 3/ Materials and Methods Prionitis lanceolata was the alga selected for study because it's associated microfauna shoued the most marked effects near the outfall. Ten stations (areas) were sampled around Pt. 42 with three replicates made at each area (Fig. 1). The samples were analyzed in themanner previously described (Clearman, 1970). Again, the Simpson andshannon-leiner indices uere used to compute values for species diversity. At Pt. Pinos four key areas in the vicinity of the outfall (areas 4, 5, and 7) and areas at the end of the point (area 10) and directly across from the outfall (area 12) were resampled (Fig. 3, Clearman, 1970). Sample analysis was carried out as befors (Clearman, 1970). The results of the original study, the summer resamples and the samples at Pt. 12 uere compared in terms of numbers of species present and the diversit indices. In order to statistically compare areas felt to be polluted with natural, unpolluted areas, the data was divided into three groups. Pt. Pinos was divided into tuo groups designated "polluted", composed of arsas felt to be heavily damaged, as shoun by lou values for the diversity indices and the small number of species found, and Funpolluted", consisting of the remaining areas around the point. These tuo groups and Pt. 42 (all areas) were compared by employing the Student's t test, using the mean values and standard deviations of the species diversity indices and the number of species of the three groups. Finally, the "unpolluted" group at Pt. Pinos and the areas of Pt. 42 uere randomly divided into equal groups and compared by the Student's t test, as above, to test the internal variability of the three criteria tested. Results Et. 42 The tabulated data for the samples at this point are shoun in Table 1. Table 2 shous the species composition for these samples. A scattered distribution, as expected in a natural environment, is seen. Figure 2 shous a distribution of the animals by phyla. Figure 3 is a plot of the mean number of species against sample areas. Also on thegraph are the results of the first study and summer resampling at Pt. Pinos for comparison. (Note: The identification of annelids at Pt. 42 and in the resample are more complete than those at Pt. Pinos. This accounts for an increase of 2-3 species. Houever the incresse in number of species is also due to finding types not previously found at Pt. Pinos. igure h is a plot of mean values of thesimpson index against sample areas, uhile Figure 5 does the same for the Shannon-einer diversity index. As before, Pt. Finos results are incorporated for comparison. Note that in all cases the values at Pt. 42 are higher than those at Pt. Pinos, and that the variations in these values at Pt. 42 are not as extreme as those at Pt. Pinos. t. Pinos Table 3 shous the tabulated data for these samples, uhile able 4 shous the species composition. Note the similarity to the results of the original survey. Figure 6 shous the distribution by phyla, and Figures 3, 4 and 5 contain the mean values by areas for numbers of species, Simpson and Shannon-Meiner indices as mentioned above. The results of the summer re¬ samples correspond to those observed earlier at Pt. Pinos, especially in the area of the outfall. Areas 10 and 12 have louer values for the divesity indices and number of species than corresponding samples from the Spring. This differences seemsto be due to seasonal variation at Pt. Pinos. Table 5 presents the mean values and standard deviations derived from the tabulated data andused in comparing the three sets of samples. It alst gives the results of the comparisons betusen the groups using the Student's t test. 2/6. 21 Discussion The results shoun in Figures 3, 4 and 5 indicate that algal-microfauna associations at Pt. 42 are characterized by having more species of animals and higher diversity indices values than those at Pt. Pinos. If ue accept the idea of Pt. 12 being a control, a natural environment that is not polluted as there is reason to believe Pt. Pinos is, then this data suggests that the entirety of Pt. Pinos is somehou changed. Differences in diversity indices and numbers of species found in the samples taken around the point shou areas of changed community structure, uhich correlated with the distribution of seuage effluent imply that parts of Ft. Pinos are polluted. Accepting the hypothesis that louer values for species diversity indices and numbers of species uhen correlated with seuage effluents indicate pollution damage, Figures 3, 4 and 5 imply that all of Pt. Pinos has suffered pollution damage. Houever, to test the validity of the data statistical methods were em- ployed. Significant differences (pe.001) uere found betueen the "polluted' and "unpolluted" areas at Pt. Pinos and betueen "polluted" areas at Pt. Pinos and areas at Pt. 12.for the numbers of species of animals found associated with the algae. There was not a significant difference betueen the "unpolluted" group and Ft. 42 for this criterion, suggesting that numbers of species of animals found in a group of samples can reveal changes in a community. For the diversity indices significant differences (pe.001) were found betueen all comparisons. This uould suggest that diversity indices vary too much from sample to sample to serve as effective indicators. Houever, by comparing the "unpolluted" group with itself and the Et. 42 group uith itself insignificant differences (p2.2) were found for both diversity indices. This indicates that the igdices are consistent within a given geographical area such as Pt. Pinos and that they do serve as indicators of change in a community. The data thus implies that all oft. Finos is significantly different from the "control" area and that all of Pt. Pinos may be subtly altered by seuage pollution, aside from the heavily damaged area in the immediate vicinity of the outfall. It may be that extreme differences in the numbers of species found, as at Pt. Pinos, indicate severe changes in a community, uhile the slight dif- ferences observed at Pt. 42 are the result of natural variation. Diversity indices on the other hand may reflect more subtle changes that differences in the numbers of species do not shou. Besides helping to confirm some of the interpretations of the results of the first study at Pt. Pinos, namely that certain areas of the point are severely damaged by seuage pollution, these results allou for the formation of a hypothesis for testing for pollution damage in an area. If, by comparing algal microfauna in a presumed polluted area and a similar unpolluted or less polluted area, an investigator can demonstrate statistically significant (pe.Ol) differences in the number of species found or in diversity indices that characterize the community, then he may label his presumed area as being polluted. Houever, there are several limitations to such an hypothesis. The investigator should have some reason for presuming an arsa to be polluted- visible differences betueen the tuo areas, a polluting element in the environment or a mechanism to account for the pollution. Also, natural variations baced on environmental and seasonal differences must not be discounted As a case in point, consider the approach to the Pt. Finos area. Visible dif. ferences in the macrofauna and flora in areas near and auay from thebutfall prompted the investigation of the microfaunal communities. Significant dif ferences in the numbers of species found and in diversity indices betueen areas near the outfall and those suay from it uere found. Based on the presence of a 216 seuage outfall and a current system that would distribute the effluent in pattern corresponding to the damaged areas, the immediate outfall area was Labelled polluted. Later, in thesummer, another point was sampled and compared with Pt. Pinos. Also, Pt. Finos was resampled to determine the effects of seasonal variation on the communities. Results indicated that natural variation was less than variations observed at Pt. Pinos, andtthat Pt. Pinos algal- microfaunalassociations uere significantly different than those at a contro point. Seasonal variation over a tuo month period was found to have little effect on the Pt. Pinos microfaumal commumities. Literature Cited Clearman, David A. 1970. The Effect of Primary Treated Seuage on the Distribution and Abundance of Microfauna on Endocladia muricata, Prionitis lanceolata and Corralina vancouveriensus. Unpublished paper. 1. Location of Prionitis lanceolata sample site. around Pt. 42, Monterey, California. leplicate samples are grouped into areas. sle: 1 inch-approximately 12 meters. CHAMPION LINE NO. 810-3 CROSS SECTION -10 SQUARES TO INCH AREA AREA Table 1. Tabulated data for samples from Pt. 63 5.8 5152 6. 86 8.7 200 60 6.56 80 7.54 120 7.01 328 2 6.7 214 b. 65 136. 14 10.2 212 9.3 326 10 9.0 238 12 6.783 116 7.4 134 5 2 6. 96 10 12 1.78 1.0 2.9 2.45 2.6 1.7 3.4 2.7 2.9 3. 46 3.5 2.9 2.86 3.6 2.5 — 1.28 2. 2.12 2.0 2.64 2.3 2.6 2.53 2.4 1.9 2.1 1.2 159 6.7 234 2 7.2 84 6.8 5.65 192 7 2 5.3 110 10 86 6.26 186 5.5 2 4.3 66 144 6.7 293 11.0 144 7.6 9 2 8.3 223 12 7.13 216 10 2 7.0 194 10 86 5.8 58 2.1 2.26 2.1 2.8 2.6 2.97 2.8 2.2 1.97 2.3 1.1 1.7 1.64 2.2 0.8 0.6 2.2 2.16 2.2 1.52 able 2. Species composition for Prionitis lanced at samples from Pt. 42. Numbers are numbers of individuals per 100 grams dry weight of algae. Species occuring in only one sample are omitted. 88 81 60 1400 36 222 88 240 112 77 113 54 18 96 112 194 232 212 18 15 22 110 36 60 22 20 24 85 275 66 70 22 12 18 72 224 76 112 14 22 14 28 36 28 11 18 17 14 16 28 80 1870 900 555 330 22 1660 550 480 900 180 550 900 1400 hho 1850 900 1700 180 1080 864 1400 948 1200 2100 600 0 176 22 11 51 17 20 14 9a. 34 11 36 32 22 110 270 400 360 360 270 440 450 420 330 210 270 210 300 180 360 700 286 232 120 360 150 22 2 27 14 68 42 12 18 22 14 18 14 28 36 27 17 18 17 18 2 — E 12 28 18 18 10 26 12 26 18 18 22 2. ig. Distri samples ion by ph la for Prionitis la 42. 2 CHAMPION LINE NO. 810-3 CROSS SECTION :10 SOUARES TO INCH — — — — — L a — THSIAAO 0 LE L L LE a o eo 1 L NVU5 SON ig. 3. Plot of the mean number of species against the sample areas. Solid line shous Pt. Pinos, Spring, sample results. Broken line shous Pt. Pinos, Summer, sample results. Double line shous Pt. 42 sample results. CHAMPION LINE NO. 810-3 CROSS SECTION-10 SOUARES TO INCH PL.PINOS AREA L — L L 2 — 126 12 13 0 2 Fig. 4. Plot of mean Simpson Diversity Index against sample areas. Solid line shous Pt. Pinos, Spring, sample results. Broken line shous Ft. Pinos, summer, sample results, Double line shous Ft. 12 sample results. CHAMPION LINE NO. 810-3 CROSS SECTION -10 SQUARES TO INCH PI.PINOS II L a 12 AREA 10 12 10 83 i9. 5. Plot of mean, Stannon-Weiner Diversity Index against sample areas. Solid line shous Pt. Pinos, Spring, sample results. Broken line shous Pt. Pigos, summer, sample results. Double line shous Pt. 42 sample results. 2 L L L O 12 AREA CHAMPION LINE NO. 810-3 CROSS SECTION : 10 SQUARES TO INCH PI.PINOS PI. II 2 233 Table 3. Tabulateddata for onitis lanceolata from Pt. Pinos (summer) samples 2 7.8 9.1 50 8.2 7.7 6.9 22 2.6 4.9 2.6 86 3.6 5.3 100 4.8 4. 2 1.6 2.6 2.9 2.1 2.2 1.2 2.4 1.2 0.7 2.0 2.5 2.0 0.( 0.7 2.7 1.0 2.0 0.4 0.0 2.4 0.8 0.4 10 2 12 2 7.67 8.53 7.6 9.3 6.54 9.35 6.68 8.91 202 14 14 268 230 210 250 190 1.5 2.2 1.67 1.7 2.1 1.28 1.45 0.78 1.41 1.56 3 Table 4. Species composition for Pric mitis lanceolats samples from Pt. Pinos (summer). Numbers are numbers of individuals per 100 grams dry weight of algae. Species occuring in only one sample are omitted. 236 12 5 240 180 120 300 120 360 360 1o0 240 600 440 220 140 120 Lho 360 260 6 18 16 18 24 12 12 33 24 18 27 16 12 12 12 22 12 16 16 12 24 12 16 11 24 8 1500 1100 1330 1550 2400 1660 2400 1656 8a. 12 12 12 18 12 12 12 36 24 11 24 24 12 12 Fig. Dist (summer lancet amp. CHAMPION LINE NO. 810-3 CROSS SECTION-10 SQUARES TO INCH O — — — L THSIA AaO — Wva O + O e0 L — L L L L —L S01 285 able 5. Mean values and standard deviations for numbers o species and diversity indices for the three sreas compared employing Student'st test. Top: Mean values and standard deviations for 12 and "Polluted" and "Unpolluted" areas Pt. Finos. Means values and standard deviations for Middle: randomly divided areas of Pt. 12 and "Unpolluted" areas of Pt. Pinos. Bottom: Results of comparisons of areas using Student'st test. Point Pinos Polluted Unpolluted Point +2 (P2) Areas: 4,5,6,7,8 2,3: 9-13 1-10 No. Species Mean 3.19 8.77 9.03 Std. Dev. 0.75 2.17 1.77 Simpson Mean 1.36 2.86 2.38 Index Std. Dev. 0.68 1.13 0.91 Shannon-W Mean 0.685 1.4g Index Std. Dev. 0.29 0.76 0.41 Unpolluted Point +2 P2 P22 Simpson Mean 2.69 2.44 3.09 2.31 Std. Dev. 0.26 Index 0.301 2.5 0.305 Shannon-W) Mean 1.80 1.99 1.88 1.71 Index Std. Dev. 0.11 0.53 0.16 0.16 Results of Student'st Tests Species Simpson Index Shannon-Weiner Index PxU t-2.53 pc.001 PxU t =10.21 po.001 t-54.37 p.001 PxP2 t -5.21 po.001 PPxP2 t =18.29 p5.001 PxP2 t=243.2 pS.001 UxP2 t - 4.19 p5.001 UxP2 t- 0.1282p2.99 UxP2 t = 22.07 pe.001 XU, t - 0.492 .3ep.l U,XU2 t - 0.762 .5-p..6 P2 xP2, t - 0.637 .h-p-.5 P21xP2, t - 1.507 .74p-.8