Temperature and metabolic rate in sedentary fish from the Antarctic,North Sea and Indo-West Pacific Ocean

Resting metabolic rate was measured in demersal stages of the teleostNotothenia neglecta Nybelin from the South Orkney Islands, Antarctica, from 1985 to 1987. The relationship between and body mass (Mb) conformed to the general relationship , wherea is a proportionality constant andb is the scaling exponent. (mg O₂ h^–1) was found to scale toMb ^{(0.82±0.011)} in the summer (November to April, 1.6 to 1 850 g,n=56) and toMb ^{(0.76±0.013)} in the winter (May to October, 0.9 to 1 850 g,n=57) (values ofb are means ± SD). Although the scaling exponents were significantly different (P<0.01), was similar in the juvenile stages of summer- and winter-caught fish matched for body mass. The effects of activity on oxygen consumption was studied using a Brett respirometer. Adult stages had a factorial aerobic scope for activity of 5.7, which is similar to that reported for demersal fish from temperate latitudes. The effects of temperature on resting metabolism was investigated in fish with similar sedentary lifestyles from the North Sea (Agonus cataphractus andMyoxocephalus scorpius) and the Indo-West Pacific (Paracirrhites forsteri, P. arcatus, Neocirrhites armatus andExallias brevis). Extrapolated values of for the tropical species approached zero at 5 to 10°C. For a standard 50 g fish, for the tropical species at 25°C was in the range 3.4 to 4.4 mg O₂ h^–1, compared with 1.3 mg O₂ h^–1 forNotothenia neglecta at its acclimation temperature. Thus, the maximum metabolic rate of sedentary tropical species at 24°C is likely to be 2 to 4 times higher than inN. neglecta at 0°C. This suggests that the energy available for sustained activity is significantly lower in cold- than in warm-water fish.     

Oxygen consumption inNeomysis americana (Crustacea: Mysidacea), and the effects of naphthalene exposure

Oxygen uptake ( ) of individual mysids was measured in a novel continuous flow respirometer for 24-h periods, and in a sealed chamber respirometer for several hours. Mysids were acclimated 30–100d under conditions which allowed complete life-cycle cultivation. was normalized for mysid size using an unconventional linear regression: log ( ) vs log (body water). Compared to the conventional regression, log ( ) vs log (body mass), the new approach showed two advantages: significantly greater coefficient of determination,r ², and removal of differences in between sexes. Minimum rates of oxygen uptake measured in the continuous flow respirometer were lower than sealed chamber measurements, and may more accurately reflect standard metabolic rate. Hourly data also showed a daily cycle when adjusted to represent a standard size mysid. Exposure to ca 1.1 mg l^-1 naphthalene increased and the range of spontaneous shown by the mysids; exposure to ca 0.1 mgl^-1 naphthalene depressed the range of . Both exposures caused aberrations in the daily cycle.     

Oxygen and carbon dioxide exchange in the polychaete Nereis virens: influence of ventilation activity and starvation

The influence of ventilation activity and starvation on O₂ uptake and CO₂ production in the polychaete Nereis virens Sars was investigated during September 1986 using worms collected on intertidal flats outside the Nærå Strand estuary, Denmark. The activity level, measured as ventilation rate, , was linearly related to active O₂ uptake, , for worms inhabiting artificial burrows (i.e. tubes) in the laboratory. at 16°C was two and four times the extrapolated standard O₂ uptake ( =0) when was 100 and 500 ml g^-1 h^-1, respectively. The use of tubes had no significant effect on N. viren's temporal ventilation pattern, D_v, compared to natural burrows in sediment. The ventilation rate, , however, was 2 to 3 times higher in tubes than in burrows. Starvation affected both ventilation and gas exchange. increased slightly during the first six days remaining constant thereafter. D_v, on the othe hand, decreased during the first six days followed by a gradual increase to Day 20. The observed decrease in D_v (%) from Day 0 to 1 was attributed to insufficient acclimation of the worms. The time integrated ventilation rate, × D_v, increased gradually throughout the 20 d starvation period. and declined rapidly during the first days of starvation. After 10 d the gas exchange reached steady levels at about 50% of the initial rate. The observed decrease in metabolic rate during starvation was explained by a rapid initial reduction in specific dynamic action (SDA) followed by a gradual exhaustion of readily available metabolic reserves, e.g. glycogen and lipids.     

Ecophysiology of the benthic amphipod Monoporeia affinis in an open-sea area of the northern Baltic Sea: seasonal variations in oxygen consumption and ammonia excretion

Seasonal variations in the oxygen uptake rate ( ) and ammonia excretion rate ( ) of the benthic deposit-feeding amphipod Monoporeia affinis vere studied in an open-sea population of the northern Baltic Sea during three consecutive years (1991 to 1993). In the early summer an increase in the weightspecific by 22% (standard 1-mg individual) was registered; an even more striking change was noted in the , which showed an increase in the early summer of ca. six times the extremely low winter/spring valnes. The metabolic changes observed could not be attributed to temperature, which remained at steadystate throughout the year, nor could seasonal variations in the biochemical composition of the amphipods explain the results. Therefor, it is evident that quality-food abundance following the sedimentation of the spring phytoplankton bloom induces the observed early-summer elevations in and . In turn, the marked decrease in the towards autumn reflects the gradual depletion of quality-food in the benthic environment. Atomic O: N ratios indicated that at the sampling location, M. affinis uses lipid almost exclusively for metabolic energy throughout the year, except in early summer when the use of lipid and protein was observed to be almost equal. Results also indicated a time-lag between sedimentation of the phytoplankton bloom and metabolic response in the amphipod population.     

Genetic population structure and gene flow among deep-sea amphipods,Abyssorchomene spp., from six California Continental Borderland basins

Studies of geographic population variation needed to estimate gene flow are lacking in deep-sea biology. Using allozyme electrophoresis, I have studied population-level geographic variation among scavenging lysianassoid amphipod populations (Abyssorchomene spp.) inhabiting deep-water basins of the Southern California Continental Borderland. Samples were collected from November 1987 to November 1990, using baited traps, from six basins whose bottom depths ranged from ca. 1000 to 2100 m. Five basins (San Diego Trough, Santa Catalina, San Nicolas, Santa Cruz, Tanner Basins) could be grouped together as shallow-sill basins, with physical conditions distinctly different from a single deepsill basin (San Clemente Basin). Amphipods tentatively identified asAbyssorchomene sp. 1 collected from the shallow-sill basins were morphologically discriminated from those collected in the San Clemente Basin, which were identified asAbyssorchomene sp. 2. Results from eight enzyme loci revealed significant genetic differentiation [Nei's genetic distance (D)>0.155] of deep-sill basin-dwellingAbyssorchomene sp. 2 vsAbyssorchomene sp. 1 from the shallow-sill basins and low levels of gene flow (migration rate, <1). Comparisons of benthic fauna suggest the presence of an abyssal-related assemblage in the deep-sill basin isolated from the northern shallow-sill basins. Genetic distances among the five shallow-sill basin populations ofAbyssorchomene sp. 1 were very low (D < 0.003). Estimates of gene flow among these populations were very high ( > 24 to 170) and suggested weak isolation by distance.     

Abundance and biomass of macrobenthos in the vicinity of Carson Submarine Canyon,northwest Atlantic Ocean

The abundance, biomass and mean weight of macrofauna in the Carson Canyon region (Lat. 45°30N, Long. 48°40W) of the Grand Banks were estimated from 40 quantitative infaunal samples taken at 15 stations in June 1980. In contrast to other areas, there were no significant differences in these values between samples taken in and outside of the canyon. Abundance ( = 716 individuals m^-2), biomass ( = 10.9 gm^-2), and mean weight ( = 0.03 g individual^-1) displayed no consistent relationship with depth, but stations grouped according to sediment type had differing dominant taxa. Biomass and abundance at the edge of the Grand Banks are unexpectedly low. Larger animals, including fishes, that forage there must depend more on pelagic than on benthic food sources.     

Influence of food concentration on the physiological energetics and growth ofOstrea edulis larvae

Feeding, respiration and growth rates of oyster (Ostrea edulis L.) larvae reared at five food levels were measured throughout the entire larval period. Energy budgets were derived as a function of alga (Isochrysis galbana Parke) food concentration. Ingestion rate (IR, cells h^-1) and oxygen consumption rate ( , nl h^-1) were almost isometric functions of larval size [ash-free dry weight, (AFDW, g)], characterized by the equations: IR=803.9 AFDW^1.13 and =4.85 AFDW^1.09. Ingested ration was directly correlated to cell concentration up to a maximum at 200 cells l^-1, with further increases failing to support higher ingestion rates. Likewise, growth rate linearly increased with food ration up to 100 cells l^-1 (max. growth efficiency,K ₁=25%) and reached a maximum at 200 cells l^-1 (growth rate=5.6 m d^-1), with further increases in food not supporting significantly faster growth. Maintenance ration was 2 to 3% daily dry weight (DW); optimum ration increased during larval development from 5 to 20% DW; maximum ration was 20% DW. During larval rearing, an increasing feeding schedule of 50, 100 and 200 cells l^-1 from Days 0, 5 and 10, respectively, is recommended.     

Growth,condition and specific dynamic action in the mussel Mytilus edulis recovering from starvation

Juvenile Mytilus edulis were grown individually in plastic racks in a tidal salt marsh for 72 d in 1984, starved in the laboratory for 130 d, and then fed the alga Isochrysis galbana daily for 64 d. Oxygen consumption was measured at various times during the course of starvation and recovery. The effects of both size (tissue dry weight) and condition (tissue size relative to shell size) on the rate of oxygen consumption during the course of starvation and recovery were analyzed by multiple regression. Weight-specific preprandial rate was inversely correlated with both size and condition. Weight-specific active rate (measured shortly after feeding) was correlated with size but not condition. Relative Scope for Activity was inversely correlated with size and positively correlated with condition. Relative Specific Dynamic Action (RSDA; the integrated physiological and mechanical response to a meal) was initially correlated negatively with size and subsequently positively with condition. Glycogen content was shown to be positively correlated with condition in mussels before starvation and during recovery. During recovery, experimental mussels returned to 90% of their estimated dry weight prior to starvation, and from 53% after starvation. At weekly intervals during recovery, oxygen consumption was measured following a meal until it returned to the preprandial rate. Both pre- and postprandial volumes (l at STP) of oxygen consumed per hour ( O _2pre and O _2post , respectively) increased significantly during recovery. The postprandial rise in oxygen consumption increased significantly from 15% to 23% of O _2pre . At the end of the recovery period, RSDA [( O _2post – O _2pre )/ O _2pre ] was independent of final dry weight, but was significantly correlated with percent dry weight recovered (r ²=0.44; df=10; P<0.02). The increase in RSDA may reflect increased utilization of food and its conversion to soma during recovery from starvation, as distinct from mechanical energy expenditure (feeding activity) following a meal.     

Comparative studies on the metabolism of shallow-water and deep-sea marine fishes. IV. Patterns of aerobic metabolism in the mesopelagic deep-sea fangtooth fish Anoplogaster cornuta

Measurements have been made of oxygen consumption rates O₂ of 10 specimens of the mesopelagic deep-sea fangtooth fish Anoplogaster cornuta. Determinations were made at 1 atm pressure, at temperatures of 3°, 7°, and 10°C, at dissolved oxygen concentrations ranging from near saturation to zero, with the fish swimming at low, controlled speeds. Weight-specific O₂ were uniformly low. They showed Q₁₀'s of 2.5 and 1.3, respectively, in the temperature ranges 3° to 7°C, and 7° to 10°C, at dissolved oxygen concentrations above 2 ml (standard temperature and presusure, STP)/1. Measurable O₂ continued in these fish at dissolved oxygen concentrations down to the lowest levels detectable with our instruments. At 7°C the average critical oxygen tension (P _c ) for the entire group was near 35 mm Hg. However, there is a statistically significant positive slope to the regression line relating O₂ to P _c for individual fish. The physiological and ecological significance of these results is discussed, particularly with reference to thermal effects and to the basis for survival by A. cornuta in the oxygen minimum layers of the eastern Pacific Ocean.     

Quantitative estimates of the meiofauna from the deep sea off North Carolina,USA

In 1974 the authors collected, sorted and enumerated meiofauna from 400, 800 and 4000 m off North Carolina, USA. Samples were replicated respectively with 4 boxcores and 21 subsamples, 2 boxcores and 7 subsamples and 2 boxcores and 8 subsamples. Total meiofaunal numbers were highest in fine silt sediment at 800 m ( =891.9 10 cm_–2) and lowest in very fine silt at 4000 m ( =73.5 10 cm_–2). Fine sand at 400 m yielded a mean of 442.4 10 cm^–2. At all depths, most fauna were located in the upper 3 cm of sediment ( depth distribution=2.2 cm), and typically only nematodes and foraminiferans were found below 4 cm. Total community abundances significantly differed with depth; however, there were no differences among replicate boxcores at particular depths. Since most (85.7%) of the variance was associated with subsamples from a boxcore, it appears that meiofauna densities are homomeneous within large areas at particular depths, and that patchiness is a smallscale phenomenon at the level of the 10 cm² subsampler. Comparisons of sorting efficiencies of live and preserved samples indicated that to accurately enumerate formainiferans, samples must first be fixed and stained, while turbellarians and oligochaetes must be sorted live.Contribution No. 151 from the Belle W. Baruch Institute for Marine Biology and Coastal Research. This research was supported by the Oceanography Section, National Science Foundation, NSF Grant GA-42792, and GC-00005 to Duke University for R.V. Eastward operations.     

Thallium in marine plankton

Concentrations of thallium in phytoplankton (0.02 to 0.8 g g^–1), zooplankton (0.03 to 0.5 g g^–1) and ichthyoplankton (0.1 g g^–1) from the central Pacific were comparable , as were the atomic ratios of thallium to calcium (3x10^–6) and to potassium (1x10^–6) in those organisms. These relatively constant ratios, plus the biounlimited ocean profile of thallium, indicate that it is rapidly cycled through plankton in the same manner as potassium, its principal biogeochemical analogue. The higher atomic ratios of thallium to potassium in pelagic clays (6x10^–6) and ferromanganese nodules (4x10^–3) suggest that both biological transport processes and abiotic transport processes influence this trace element's oceanic cycle.     

Comparative studies on the metabolism of shallow-water and deep-sea marine fishes. V. Effects of temperature and hydrostatic pressure on oxygen consumption in the mesopelagic zoarcid Melanostigma pammelas

Measurements have been made of routine oxygen consumption rates ( ) of the mesopelagic deep-sea zoarcid fish Melanostigma pammelas. Determinations were made over ecologically relevant ranges of 3 variables; temperature (3° to 10°C), hydrostatic pressure (1 to 170 atm), and oxygen partial pressure (1 to 160 mm Hg). Weight-specific s were uniformly low. Of the 3 test variables, only temperature had significant metabolic effects within the ranges studied. Q₁₀'s were 6.75 between 3° and 5°C. 1.47 between 5° and 7°C, and 17.4 between 7° and 10°C. These Q₁₀'s were constant over the hydrostatic pressure range studied. Between 3° and 7°C the fish regulated their rates of oxygen consumption down to PO₂'s comparable to those occurring in their natural environments (6 to 12 mm Hg). The showed no capacity to tolerate anoxic conditions. The physiological and ecological significance of these results is discussed, particularly with reference to thermal effects and to the basis of survival of this fish in the oxygen minimum layers of the eastern Pacific Ocean. Since it is possible to maintain M. pammelas in the laboratory for extended periods of time (over 12 months) it could serve as the basis for many interesting studies of deep-sea fish biology.     

Genetic differentiation between spatially-disjunct populations of the deep-sea,hydrothermal vent-endemic amphipod Ventiella sulfuris

Species endemic to deep-sea hydrothermal vent ecosystems have disjunct distributions imposed by the island-like arrangement of their specialized habitats. Using allozyme electrophoresis, we examined genetic population structure of the hydrothermal vent amphipod Ventiella sulfuris Barnard and Ingram, 1990. Samples from five sites along the East Pacific Rise (EPR) and two along the Galapagos Rift were collected in 1990 and 1988, respectively. Variability, based on 12 enzyme loci, was relatively high (proportion of polymorphic loci whose most common allele not greater than 0.95 in frequency) =41.6%; (mean heterozygosity) =0.158] compared with shallow-water marine and freshwater amphipods, and similar to the deep-sea lysianassid Eurythenes gryllus. Genetic divergence among populations spread along a cöntiguous rift axis (i.e., EPR) was low [Nei's genetic distance (D) ranged from <0.001 to 0.018]. Genetic tructure analysis suggests that along a contiguous ridge axis migration occurs in a stepping stone manner and is unconstrained by distances as great as 1200 km (migration rate, , ranged from 1.9 to 67.8 ind. generation^–1). However, genetic divergence between populations on disjunct ridge axes was extremely high (D ranged from 0.438 to 0.476). Most of the variance in gene frequencies was due to the differences between the major subpopulations inhabiting the two distinct ridge axes, EPR and Galapagos Rift. Apparently, very little migration and gene flow occur between these major subpopulations (M1). This level of genetic divergence may be sufficient to justity separation of EPR and Galapagos Rift populations at the species level. Futher analyses of morphological characters is required before taxonomic status can be assigned.     

Light absorption and utilization among hermatypic corals: a study in Jamaica,West Indies

The chlorophyll specific absorption coefficient ( _c) was measured for zooxanthellae from six hermatypic coral species obtained, where possible, from four depths (1, 10, 30, 50 m) on reef sites near Discovery Bay, Jamaica in February and March 1983. Measurements of photosynthetic rates versus irradiance, as well as cellular and areal chlorophyll a, were also performed on these colonies or sister colonies. Together the data were used to compare minimum quantum requirements (1/Φ m) among species and depths and to assess the importance of light utilization to the growth and depth distribution of these corals. Our data suggest that, although _c was found to decrease with depth, interspecific differences in _c do not occur for zooxanthellae from the corals investigated. Minimum quantum requirements (1/Φ m) decreased significantly with depth, thereby reflecting an increase in photosynthetic light utilization efficiency with decreasing irradiance. Interspecific differences in 1/Φ m determinations were suggested but not statistically conclusive. We conclude that interspecific differences in gross photosynthesis, and perhaps growth and depth distribution, are primarily attributable to differences in the light utilization capacity of the whole coral, as reflected by the product of _c ^′ and chlorophyll per unit surface area, and in-situ quantum efficiencies. This research was performed under the auspices of the US Department of Energy under Contract No. DE-AC02-76CH00016     

Trophic variation with length in two ommastrephid squids, <Emphasis Type="Italic">Ommastrephes bartramii</Emphasis> and <Emphasis Type="Italic">Sthenoteuthis oualaniensis</Emphasis>

From 1998 to 2001 a total of 200 Ommastrephes bartramii (27 paralarvae) and 170 Sthenoteuthis oualaniensis (14 paralarvae) were sampled from the Central North Pacific. One group of non-paralarval O. bartramii (n = 30) was sampled from farther northwest in 1996. The δ¹⁵N of mantle muscle of non-paralarval O. bartramii ( = 12.4‰) was significantly greater than that of non-parlarval S. oualaniensis ( = 8.1‰) (P < 0.001). The δ¹⁵N of whole paralarvae of O. bartramii ( = 6.4‰) was not significantly different than parlarvalae of S. oualaniensis ( = 6.1‰) (P = 0.528). There was no significant difference between the mantle muscle δ¹⁵N values of male (n = 95, = 13.3‰) and female (n = 18, = 12.9 ‰) O. bartramii greater than 300 mm mantle length (ML) (P = 0.15). There was also no significant difference between the mantle muscle δ¹⁵N values of male (n = 15, = 7.2‰) and female (n = 26, = 7.3 ‰) S. oualaniensis in the same size range (P = 0.41). Overall there was a distinct logistic increase in δ¹⁵N with mantle length for O. bartramii, whereas S. oualaniensis showed an exponential increase in δ¹⁵N with mantle length that was stronger within individual years than with all samples combined. In general, adult O. bartramii are more than a trophic level above S. oualaniensis (4.3‰, 1.3 TLs). Because of the nature of the sampling protocol, this study could not separate spatial and temporal effects on the δ¹⁵N signals from each squid species. This study demonstrates the ability of stable isotope analyses to differentiate trophic levels between squid species as well as track trophic changes across size ranges from paralarvae to adults. Additional research is needed to validate these trophic changes across size within individuals.     

Morphological and genetic variation in Japanese populations of the anemonefish Amphiprion clarkii

Geographic populations of the anemonefish Amphiprion clarkii (Bennett) from 6 widely separated locations off the coast of southern Japan are morphologically different, exhibiting (1) latitudinal clinal patterns in color pattern and meristics and (2) degrees of genetic differentiation. Electrophoretic examination of 7 polymorphic loci (95% level) among 6 populations collected between July 1979 and January 1980 revealed an average genetic distance value of 0.008. Significance was demonstrated for 39 out of 105 (37%) tests of heterogeneity. Averaged over the 6 populations, the percentage of polymorphic loci ( _0.95) was 24.6% and the proportion of heterozygous loci per individual ( ) was 0.0613. The results indicate that genetic clines are absent and suggest that the morphological variation may not be genetic. Relative isolation of genetic populations may be maintained by (1) localized larval dispersal resulting from a relatively short larval stage, and (2) current gyres tending to trap larvae, increasing the return of juveniles to their adult coastal habitat. Morphological clines may be due to clines in ecological parameters related to latitude.Contribution No. 39 of the Tatsuo Tanaka Memorial Biological Station     

Statistical inference on patch-specific survival and movement rates from marked animals

Empirical estimates of patch-specific survival and movement rates are needed to parametrize spatially explicit population models, and for inference on the effects of habitat quality and fragmentation on populations. Data from radio-marked animals, in which both the fates and habitat locations of animals are known over time, can be used in conjunction with continuous-time proportional hazards models to obtain inferences on survival rates. Discrete-time conditional logistic models may provide inference on both survival and movement rates. We use Monte Carlo simulation to investigate accuracy of estimates of survival from both approaches, and movement rates from conditional logistic regression, for two habitats. Bias was low (relative bias < 0.04) and interval coverage accurate (close to the nominal 0.95) for estimates of habitat effect on survival based on proportional hazards. Bias was high ( relative bias 0.60) and interval coverage poor ( = 0.26 vs. nominal 0.95) for estimates of habitat effect based on conditional logistic regression; bias was especially influenced by heterogeneity in survival and the shape of the hazard function, whereas both bias and coverage were affected by ‘memory’ effects in movement patterns. Bias estimates of movement rate was low ( relative bias < 0.05), but interval coverage was poor ( = 0.48–0.80), possibly as a result of poor performance of a Taylor series estimate of variance. An example is provided from a radio-telemetry study of 47 wintering American woodcock (Scolopax minor), illustrating practical difficulties in field studies to parametrize these models. We also discuss extensions of continuous-time models to explicitly include a movement process, and further examine tradeoffs between continuous and discrete models.     

Flow rate and particle concentration within the house of the pelagic tunicate Oikopleura vanhoeffeni

The food-concentrating filter within the house of Oikopleura vanhoeffeni Lohmann consists of three layers, upper and lower fine-mesh layers and an intermediate coarse-mesh layer. The filter is a natural analogue of man-made tangential, or cross-flow, filters that are used to concentrate fine particulate and colloidal material by the exclusion of water. Flow analysis using video microscopy of O. vanhoeffeni, collected in 1989 and 1990 at Logy Bay, insular Newfoundland, indicates concentration of particles of 74 to 1089 times that of ambient seawater ( ,n=23), for individuals 1.8 to 5.4 mm long. The smallest individuals had the highest concentration factors. Flow through the house is predominately affected by viscous forces and values of Reynolds numbers are 1. Flow rates through the house of O. vanhoeffeni indicate a maximum filtration capacity of ca. 1.4 to 26 liters d^-1 and clearance rates (assuming ca. 50% of the time is spent not actively pumping) of 0.7 to 13 liters d^-1, depending on body size. The estimated thickness of the boundary layer around individual fibers of the food-concentrating filter is 0.08 m, which is less than the pore width. The estimated concentration factors suggest that colloidal and fine particulate material may aggregate into larger particles within the food-concentrating filter and food tube during feeding.     

Genetic parentage in the indigo bunting: a study using DNA fingerprinting

Summary Parentage of nestlings in a North Carolina population of indigo buntings (Passerina cyanea) was analyzed using DNA fingerprinting. Three minisatellite DNA probes (wild type M13, Jeffreys' 33.15 and 33.6) were used to analyze nuclear DNA isolated from muscle biopsies of 63 nestlings, their parents, and other local adults. Each probe detected approximately 15 scorable fragments per individual, with 18%–39% overlap between probes. The proportion of bands shared (using all fragments over all three probes) between presumably unrelated adults averaged 0.23. Of the 63 offspring analyzed, 35 had at least one fragment not present in either putative parent. The distribution of offspring with novel fragments was distinctly bimodal. The lower mode (offspring with 0, 1, or 2 novel fragments, N=41) fit a Poisson distribution, a pattern expected if mutation (estimated rate per fragment= 0.01) were the source of the novel fragments. The remaining 22 offspring had more novel fragments than could be explained by mutation alone (minimum of four independent fragments across all three probes, =8.2). A low band-sharing proportion with the resident male ( =0.24) and high band-sharing with the resident female ( =0.60) implicated extra-pair fertilizations as the source of all 22. Thus in this sample, 35% of all nestlings came from extra-pair fertilizations and none from intra-specific brood parasitism. Of 25 broods sampled, 12 (48%) had at least one excluded offspring. In 3 broods all of the offspring excluded the resident male. Band-sharing proportions between excluded nestlings within a brood could not distinguish between single and multiple extra-pair paternity. Although young males tended to be excluded less often than older males, wing length and weight were not associated with the frequency of exclusion. Weight and wing length of females also were not associated with involvement in EPCs. Six of the 22 excluded offspring (in 3 broods) shared a large proportion of bands and had fewer than four novel fragments when compared to the fingerprints of a neighboring territorial male, implicating those males as actual fathers. The parentage of the remaining 16 offspring (in 9 broods) could not be clearly assigned because (1) one or more neighbors were not sampled or (2) difficulties in scoring across gels prevented confident alignment of fingerprint bands, and insufficient DNA was obtained from muscle samples to allow reanalysis of potential actual fathers on the same gel as excluded nestlings.