Simultaneous nitrification and denitrification in a mixed methanotrophic culture

Simultaneous nitrification and denitrification using a mixed methanotrophic culture was investigated. When both NO₃ ^–-N (108 mg l^–1) and NH₃-N (59 mg l^–1) were added into batch reactors, nitrate removal was complete within 10 h at the rate of 47 mg NO₃ ^–-N g VSS^–1 day^–1 when dissolved oxygen (DO) concentration was maintained at 2 mg DO l^–1. Ammonia removal started simultaneously with nitrate removal at a slower rate of 14 NH₃-N g VSS^–1 day^–1. No significant accumulation of nitrite or nitrate during ammonia utilization suggested the occurrence of simultaneous nitrification and denitrification.     

Effects of sublethal exposure to copper on diel activity of sea catfish,Arius felis

The effect of sublethal exposure to copper on the diel activity of sea catfish, Arius felis, was examined after 72 h static exposure to copper (0.0, 0.05, 0.1, or 0.2 mg l^–1 ). The locomotor behavior of 28 individual fish was electronically monitored in a 16-compartment rosette tank. Activity was quantified by the number of entries into compartments per hour. Controls and copper-exposed fish (0.05 mg l^–1) behaved similarly after treatment and displayed no significant effects of handling or copper exposure on diel activity. However, fish exposed to 0.1 and 0.2 mg l^–1 copper were hyperactive immediately following exposure; these fish had significantly greater numbers of entries into compartments per hour for entire 24-h monitoring periods (P < 0.01) and for the 12 h of photophase (P < 0.001) and of scotophase (P < 0.001) than fish in the other two experimental groups. The normal diel activity cycle was abolished. Additionally, exposure of fish to copper at 0.1 and 0.2 mg l^–1 resulted in significantly less variability in activity (P < 0.05) compared to control fish and those exposed to 0.05 mg Cu l^–1.     

Mercury in the marine food chain in the Southern Ocean at Macquarie Island: an analysis of a top predator,Patagonian toothfish (<Emphasis Type="Italic">Dissostichus eleginoides</Emphasis>) and a mid-trophic species,the warty squid (<Emphasis Type="Italic">Moroteuthis ingens</Emphasis>)

The characteristics and habitat of the Patagonian toothfish (Dissostichus eleginoides) are typical of fish that accumulate high concentrations of mercury. In this study, mercury determinations were made on samples of muscle tissue from Macquarie Island toothfish and the Southern Ocean deepwater warty squid (Moroteuthis ingens). The analysis of mercury in the biological tissues was made by cold vapour-atomic absorption spectrometry following acid digestion. Performance of the analytical procedure was assessed by analysis of certified reference material (DORM-2, dogfish muscle). Mercury concentrations of 16 Macquarie Island toothfish ranged from 0.12 mg kg^–1 (550 g, 381 mm TL) to 0.59 mg kg^–1 (6,100 g, 823 mm TL), with a mean concentration of 0.33±0.12 mg kg^–1. A significant correlation was found between mercury and either toothfish weight or total length. The fish analysed were juveniles, which suggests that larger individuals would have higher mercury concentrations well exceeding food standard code limits for mercury in fish (typically 0.5 mg kg^–1). Warty squid, also from around Macquarie Island, had a low mean mercury concentration of 0.086 mg kg^–1 in mantle tissue; no significant correlation existed between mercury concentration and either squid mantle length or total weight. It is postulated that the squid have a mechanism, possibly involving the digestive gland, that prevents bioaccumulation of mercury in the mantle, and presumably other body tissues.     

Nitrification rates in a saline medium at different dissolved oxygen concentrations

A kinetic study was carried out in a saline medium to assess the effect of O₂ on the two-step nitrification process: for the first nitritation step, 2–26 mg dissolved O₂ (DO) l^–1 was used and for the second nitratation step, 0.5–24 mg DO l^–1 was used. Nitritation rate was measured in the presence of sodium azide so as to inhibit nitratation. Ammonia-oxidizing (AOB) and nitrite-oxidizing (NOB) bacterial in the mixed culture were determined by 16 S rRNA hybridization. The affinity constants for oxygen O₂ of the AOB and the NOB were 1.66 mg O₂ l^–1 and 3 mg O₂ l^–1 respectively. The larger than the previously reported values of these constants might be due to the high salt content in the medium. High O₂ concentrations did inhibit the nitrification rate.     

Role of aquatic vegetation coverage on hypoxia and sunfish abundance in bays of a eutrophic reservoir

We studied the relation between aquatic vegetation coverage, summer dissolved oxygen and density of sunfishes (Lepomis spp.) in 10 shallow bays of a eutrophic reservoir. The bays ranged 5.2–15.7 ha in area, 0.6–1.3 m mean depth and 6–91% vegetation area coverage. Over the 10 bays mean dissolved oxygen concentration ranged 5.8–9.0 mg l^-1 in open water at least 20 m away from the vegetation, 0.8–8.1 mg l^-1 at the vegetation-water edge and 0.5–7.7 mg l^-1 in dense vegetation. Dissolved oxygen concentrations were inversely related to vegetation coverage. In open water, dissolved oxygen levels were consistently higher than 8 mg l^-1 when vegetation coverage was <20% of the bay and decreased to about 6 mg l^-1 at coverages near 80%. At the vegetation-water edge and within dense vegetation, dissolved oxygen levels dropped rapidly as vegetation coverage increased to 20% of the bay; when vegetation reached about 50% coverage, dissolved oxygen remained near 1.5 mg l^-1at the vegetation-water edge, but oxygen dropped below 1 mg l^-1in dense vegetation. Scarce vegetation harbored high Lepomis relative abundance (fish per m² of vegetation) whereas extensive vegetation harbored low relative abundance, both contributing little to absolute abundance (total fish in all vegetation); however, intermediate coverage offered a combination of mid-level fish relative abundance that together with mid-level plant coverage translated into high absolute fish abundance. We suggest this response is related to hypoxia, and where aquatic vegetation is extensive, the effect of vegetation on hypoxia and water quality in general may influence fish populations in a way similar to that often attributed to reduced foraging efficiency and increased competitive interactions.     

Efficient GA3-assisted plant regeneration from cell suspensions of three grape genotypes via somatic embryogenesis

Petioles from in vitro grown plants of interspecific grapevine hybrids cvs `Bianca', `Podarok Magaracha' and `Intervitis Magaracha' were cultured on solid NN medium supplemented with 2,4-D and BA at various concentrations. The callus developed was cultured in liquid NN medium supplemented with 0.5 mg l<sup>–1</sup> BA to induce formation of somatic embryos. Somatic embryos of globular and heart-stage developed in suspensions of `Podarok Magaracha' and `Intervitis Magaracha'. In contrast, `Bianca' did not undergo embryogenesis beyond globular stage. This made it necessary to perform subculture of the suspensions to HTE liquid medium supplemented with 0.2 mg l^–1 BA for the development of globular embryos into heart stage. Heart-stage embryos developed into torpedo-stage after subculturing suspensions of all three cultivars to liquid HTE medium supplemented with 0.1 mg l^–1 IAA and 30 mg l^–1 sodium hummate. Torpedo-stage embryo suspensions were subcultured in liquid HTE medium supplemented with 0.5 mg l^–1 BA, 0.5 mg l^–1 GA₃ and 0.5 mg l^–1 GA₃ + 0.2 mg l^–1 BA. After 12 days of incubation, plantlets were cultured on solid M2MS medium: without growth regulators and with 0.5 mg l^–1 BA. Plantlets that developed in liquid HTE media with 0.5 mg l^–1 GA₃ or 0.5 mg l^–1 GA₃ + 0.2 mg l^–1 BA produced 82–90% shoots on solid M2MS medium with 0.5 mg l^–1 BA after 50 days of culture.     

Physical and chemical post-dam alterations in the Jamari River, a hydroelectric-developed river of the Brazilian Amazon

The regulation of the Jamari River advanced peak floods by 1–2 months and increased dry-season discharges from 60 to 200 m³ s^–1, resulting in water levels approximately 1 m above those recorded before regulation. Daily variation in water level associated with fluctuations in electricity production by the dam propagated to the lower reaches of the tributary Candeias River. Dissolved oxygen (DO), temperature, pH, and conductivity measured over 1.5 years on three locations along the regulated and two free-flowing rivers indicated important alterations in the case of oxygen concentrations. DO levels shifted from saturated (7–8 mg l^–1) during the flood season (when the spillways were open releasing epilimnetic water) to hypoxic (1.5–3 mg l^–1) during the dry season (when the floodgates were closed and only hypolimnetic water used to run the turbines was feeding the river). Fluctuations in water level and variation in dissolved oxygen tended to be greater at the site closest to the dam, gradually attenuating downstream. Mitigation of the downstream effects of river regulation would require modifications in the operation of the dam.     

Uptake and accumulation of zinc in juvenile rainbow trout,Salmo gairdneri

Synopsis The toxicity of zinc to rainbow trout was determined and the 72 h median lethal concentration was found to be 2.00 mg l^–1 in freshwater, hardness 7.50 mg l^–1 as calcium. An insignificant increase in zinc concentration of internal tissues occurred in fish exposed to 1.52 mg l^–1 in freshwater for 72 h. However, there was a significant uptake of zinc by gills and the body surface. Fish exposed to 10 mg l^–1 zinc for 72 h in two-thirds sea water showed significant zinc uptake by liver, rectum and muscle, when compared to control fish. Drinking rate decreased from 1.43 to 0.26 ml kg^–1 h^–1 when zinc sulphate was added to freshwater. Trout adapted to two-thirds sea water showed no decrease in drinking, about 7 ml kg^–1 h^–1 when zinc was added to the water.     

Overturn in a hypertrophic,warm, monomictic impoundment (Hartbeespoort Dam,South Africa)

A major decrease in air temperature with an increase in wind stress created thermal instability resulting in overturn. Surface oxygen values decreased from 12.6 (170% saturation) prior to overturn to 0.4 mg · l^–1 (5.7%) after overturn. Recovery of surface oxygen concentrations to supersaturation took approximately three months. Changes in the nitrogen balance after overturn indicated that extremely high rates of nitrification occurred. Little change in phosphorus concentration was observed. Surface chlorophyll concentrations decreased from 235 to approximately 2 mg · m^–3, resulting in very low primary production values (A_max = 12.4 mg C m^–3 h^–1). Reduced rates of primary production were maintained for several months after overturn, mainly owing to the low ratio of Z_eu/Z_m (0.17). No significant changes occurred in the total bacterial population other than a redistribution within the water column. Significant changes occurred in the distribution and density of the zooplankton population. No fish kills were observed although fish populations were oxygen-stressed.     

Transport of coarse particulate organic matter in an Idaho river,USA

I investigated organic matter transport in the Henry's Fork of the Snake River, Idaho, USA, from August 1987 to November 1988. Mean discharge during the study was 15 m³ s^–1. Screens were used to sample very coarse (> 6 mm) transported aquatic macrophyte material (VCTMM). Drift nets were used to sample coarse (1–6 mm) and fine (0.25–1 mm) transported particulate organic matter (CTOM and FTOM). Mean monthly concentration of VCTMM was 0.064 mg AFDWl^–1 and was significantly higher than CTOM (0.024 mg AFDW l^–1) and FTOM (0.036 mg AFDW l^–1). VCTMM concentration was highest in December (0.163 mg AFDW l^–1) and lowest in May (0.018 mg AFDW l^–1). The sample position along a transect across the channel had a significant effect on the amount of transported organic matter collected in many months. The concentration of debris from individual species tracked the standing stock of that species during the growing season. In Fall, a dramatic increase in VCTMM corresponded to a decrease in macrophyte standing stock. FTOM and CTOM concentrations were highest in January (CTOM: 0.048; FTOM: 0.111 mg AFDW l^–1), lowest in November 1988 (<0.006 mg AFDW l^–1), were not correlated with discharge, and were inversely correlated with the standing stocks of macrophytes upstream, probably because macrophyte beds influenced the retentiveness of the channel. Standing stock of aquatic macrophytes was highest in September–October (5.2 kg wet weight m ^–2) and lowest in February (1.7 kg wet weight m^–2). Annual movement of particulate organic matter past the sampling point was about 45 000 kg AFDW, of which 21 000 kg was VCTMM, 8 000 kg was CPOM, and 16 000 kg was FPOM.     

Thermal, mixing, and oxygen regimes of the Salton Sea, California, 1997–1999

The Salton Sea is a shallow (mean depth = 8 m; maximum depth = 15 m), saline (41–45 g l^–1), intermittently mixing, 57 km long, 980 km² lake located in the arid southwestern United States. The Sea is a wind driven system, with predominant winds paralleling the long axis of the lake, being strongest in spring and weakest in summer and fall. The Sea mixed daily or nearly daily between September and January. During this cooling period, moderate to high levels of dissolved oxygen (3–11 mg l^–1) were found throughout the water column. Mean water column temperature ranged from a minimum of 13–14 °C in early January to a maximum of 30–34 °C in July–September. During most of this warming period, the Sea was thermally stratified but subject to periodic wind driven mixing events. Winds were stronger in spring 1998 than in 1997 or 1999, causing more rapid heating of the lake that year and also delaying onset of anoxic conditions in bottom waters. During summer months, mid-lake surface waters were sometimes supersatured with oxygen, and bottom waters were hypoxic or anoxic with sulfide concentrations > 5 mg l^–1. Oxic conditions (> 1 mg O₂ l^–1) often extended a few meters deeper nearshore than they did well offshore as a consequence of greater mixing nearshore. Mixing events in late summer deoxygenated the entire water column for a period of days. Consumption of oxygen by sulfide oxidation likely was the principal mechanism for these deoxygenation events. Sulfide concentrations in surface waters were 0.5–1 mg l^–1 approximately 3 days after one mixing event in mid-August 1999. These mixing events were associated with population crashes of phytoplankters and zooplankters and with large fish kills. In the southern basin, freshwater inflows tended to move out over the surface of the Sea mixing with saline lake water as a function of wind conditions. Salinity gradients often contributed more to water column stability than did thermal gradients in the southeasternmost portion of the lake.     

Population Biology and Growth of Ozark Cavefish in Logan Cave National Wildlife Refuge, Arkansas

Ozark cavefish, Amblyopsis rosae, is a threatened species endemic to the Springfield Plateau of the Ozark Highlands in Arkansas, Missouri, and Oklahoma. One of the largest known Ozark cavefish populations, located in Logan Cave, Arkansas, was surveyed 25 times over a two-year period between 1993 and 1995. During the study, 147 Ozark cavefish > 30 mm (TL) were marked with visual implant tags and 140 Ozark cavefish were available for recapture; 68 were recaptured 189 times and the rest (72) were never recaptured. Individual Ozark cavefish persisted in Logan Cave for a relatively short time. Only 14% of 80 fish tagged during a previous study in 1992 were recaptured during this study, and half of all recaptured fish disappeared within three months. However, if a fish persisted for at least seven months in the cave, its probability of being recaptured over an additional year was high. Maximum persistence of a tagged fish was 28 months, suggesting these fish have a maximum life-span of 4–5 years. Growth averaged 0.6 mm per month, with maximum recorded growth of 6 mm per month and a maximum size of 65 mm TL. Smaller fish grew faster than larger fish but growth rates were sporadic, with several mid-sized fish (45–49 mm) showing little growth (0–3 mm year^-1) while some fish > 50 mm grew up to 12 mm year^-1. Most fish gained in length during April–October, the same period a maternity colony of gray bats occupied the cave. Gross Ozark cavefish movement over the study period ranged up to 1002 m, with a mean movement of 1.2 m day^-1; movement was positively correlated with Ozark cavefish total length. Death seemed the most likely explanation for loss of tagged Ozark cavefish, including fish that emigrated out of the cave. Little up-stream movement was recorded between reaches and did not account for loss of tagged fish. Reproduction within the cave and immigration from the aquifer accounted for persistence of Ozark cavefish in Logan Cave.     

A reproducible procedure for plant regeneration from seedling hypocotyl protoplasts of Vigna sublobata L.

Viable protoplasts of Vigna sublobata L. were isolated enzymatically from hypocotyls of axenic seedlings. Protoplast yields were dependent upon seedling age, with maximum yields (2.25 ± 0.35 × 10⁶ g fwt^–1) from seedlings aged 6 d. Protoplasts regenerated cell walls and underwent sustained divisions when cultured in either agarose-solidified or liquid K8P medium. The plating density affected the division frequency and plating efficiency; the division frequency (68 ±0 6.0%) was maximum at 4.0 × 10⁴ ml^–1 while plating efficiency was maximum (1.3 ± 0.1%) at 5.0 × 10⁴ ml^–1. Dividing protoplasts developed into microcalli, which produced glossy green compact nodular calli on transfer to 8.0 gl^–1 w/v agar-solidified medium containing MS salts, B5 organic components, 30 g l^–1 sucrose, NAA (0.2–0.5 mg l^–1), zeatin riboside (0.5–2.0 mg l^–1) and GA₃ (0.5–1.0 mg l^–1). These calli, after sub-culture on the same medium, produced shoot buds which underwent elongation following transfer of tissues to 6.0 g l^–1 agar-solidified B5 medium containing 30g l^–1 sucrose, IBA (0.01 mg l^–1) and BAP (1.0 mg l^–1). Elongated shoots developed roots after transfer to 8.0g l^–1 agar-solidified, hormone-free MS medium with 30 g l^–1 sucrose.Abbreviations IAA indole-3-acetic acid - IBA indole-3-butyric acid - BAP 6-benzyladenine or benzylaminopurine - B5 medium after Gamborg et al (1968) - 2,4-D 2,4-dichlorophenoxyacetic acid - GA³ gibberellic acid - 2,i-P 6-(--dimethylallylamino) purine - MS medium after Murashige and Skoog (1962) - NAA 1-naphthaleneacetic acid     

Food consumption of Sarotherodon mossambicus (Trewaves) exposed to sublethal concentration of diammonium phosphate

S. mossambicus was exposed to toxic and sublethal concentrations of the fertilizer diammonium phosphate (0.2 to 1.0 g l^–1). Mortality, food utilization and growth were studied. At a concentration of 0.6 g l^–1 DAP, 100% mortality was observed within 96 h; no mortality occurred at 0.5 g l^–1; LC50 was 0.55 g l^–1. Rearing the fish in increasing sublethal concentrations of DAP, it was found that the feeding rate decreased from 25.4 mg g^–1 fish^–1 d^–1 (fish reared in DAP-free water) to 10.1 mg g^–1 d^–1 at the highest sublethal concentration (0.5 g l^–1). Growth rate was drastically reduced. At high sublethal concentrations of DAP, the fish lost reserve energy, in addition to the energy obtained from food intake for survival, as a result of increased swimming activity and opercular beats.     

Efficient Regeneration of Curcuma amada Roxb. Plantlets from Rhizome and Leaf Sheath Explants

Efficient and simple protocols were developed for conversion of embryos derived from microspores in rape. The frequency of embryo conversion was higher than 80% on improved media without pretreatments using ABA, GA₃ or desiccation, which had been required before embryos were transferred to solid media for conversion. While on basal media such as 1/2MS, the conversion frequency was no more than 32%. Two groups of embryos at different developmental stages, 7 mm embryos and 3 mm embryos, were used as material to select the most suitable medium. Different components were required for successful conversion of the two groups of embryos. For 7 mm embryos 1/2MS medium with Ca²⁺ concentration of 900 mg l^–1 was optimal, while for 3 mm embryos, 1/2 MS_N+V+Ca (half strength MS with 450 mg l^–1 CaCl₂·2H₂O, 100 mg l^–1 NH₄NO₃ and enhanced vitamins) was a suitable medium. The results indicate that calcium may play an important role in the conversion of embryos derived from microspores, and can replace, to some extent, the practice of drying and growth supplements, which have been widely used for maturation, and desiccation of embryos.     

Impact of cyprinids on zooplankton and algae in ten drainable ponds

To study the impact of cyprinids on algae, zooplankton and physical and chemical water quality, ten drainable ponds of 0.1 ha (depth 1.3 m) were each divided into two equal parts. One half of each pond was stocked with 0 + cyprinids (bream, carp and roach of 10–15 mm), the other was free of fish. The average biomass of the 0 + fish at draining of the ponds was 466 kg ha^–1, to which carp contributed about 80%.The fish and non-fish compartments showed significant differences. In the non-fish compartments the density of Daphnia hyalina was 10–30 ind. l^–1 and that of Daphnia magna 2–4 ind. l-^–1, whereas in the fish compartments densities were c. 1 ind. l^–1. Cyclopoid copepods and Bosmina longirostris, however, showed higher densities in the fish compartments. The composition of algae in the two compartments differed only slightly, but the densities were lower in the non-fish compartments. The significant difference in turbidity was probably caused by resuspension of sediment by carp. No significant difference in nutrient concentration between the compartments was found.     

The effect of dissolved oxygen on the growth of young-of-the-year winter flounder,Pseudopleuronectes americanus

Synopsis The effects of constant and diurnally fluctuating levels of dissolved oxygen on the growth of young-of-the-year winter flounder,Pseudopleuronectes americanus, were examined under controlled laboratory conditions. Fish were exposed for either 10 or 11 weeks to constant levels of 6.7 (high) and 2.2 (low) mg l^–1, and a diurnal fluctuation, ranging from 2.5 to 6.4 mg 0₂l^–1. Growth rates, calculated for both standard length and weight, for fish exposed to low and diurnally fluctuating levels were significantly reduced (p < 0.001) as compared to those for fish exposed to the high level. Growth rates of fish exposed to the high level were over twice those of fish held under low oxygen conditions. Under fluctuating conditions, fish grew at intermediate rates. Following these exposures, all fish were subsequently held at 7.2 mg Oz l^–1 for five weeks. Growth rates increased over two and a half times for fish previously exposed to the low oxygen level and were significantly (p < 0.001) higher than for the other two groups.     

Effects of activated charcoal, explant size, explant position and sucrose concentration on plant and shoot regeneration of Lilium longiflorum via young stem culture

An efficient system for the in vitro plant and shootregeneration of Lilium longiflorum was developed andaccomplished using transverse thin cell layers (tTCL) of young stems.tTCLs were cut transversely along young stems from which the shoot-tipshad been removed. Sections were measured accurately using a graded gridand were cut in 4 mm × 4 mm × 1 mm cubes, eliminatingepidermal tissue, and were cultured on one-half MS medium containing 8 gl^–1 agar, different sucrose concentrations (10, 20, 30 or 40g l^–1), and with or without 1 mg l^–1 activatedcharcoal (AC). Plants formed on the surface of tTCLs within 60 days onone-half MS medium containing 8 g l^–1 agar and 20 gl^–1 sucrose. Sections of 1 mm taken just below the apicalarea developed buds within 15 days, whereas the sections closer to thebase required about 45 days. Shoot regeneration was enhanced whensucrose concentration was used at 30 or 40 g l^–1 after 60days of culture. No root formation occurred. Both shooting and rootingoccurred when sucrose was used at 20 g l^–1. The plantletswere transferred to soil and grew well under greenhouseconditions.     

Turbidity enhances feeding abilities of larval Pacific herring,Clupea harengus pallasi

Fishes inhabiting estuaries, rivers, and embayments are subject to turbid conditions. Larvae of many fishes utilize estuaries as nursery areas. For visual plankton feeders such as larval fishes, turbidity may reduce search and reaction distances, resulting in lowered feeding abilities. In this study feeding Pacific herring larvae, Clupea harengus pallasi, were exposed to suspensions of estuarine sediment and Mount Saint Helens volcanic ash at concentrations ranging from 0 mg · l^–1 to 8 000 mg · l^–1. In all experiments, maximum feeding incidence and intensity occurred at levels of suspension of either 500 mg · l^–1 or 1000 mg · l^–1 with values significantly greater than controls (0 mg · l^–1). Feeding decreased at greater concentrations. The suspensions may enhance feeding by providing visual contrast of prey items on the small perceptive scale used by the larvae. Larval residence in turbid environments such as estuaries may serve to reduce predation from larger, visual planktivores, while searching ability in the small larval perceptive field is not decreased.     

Age and growth estimates of the bull shark,Carcharhinus leucas,from the northern Gulf of Mexico

Synopsis Length at age and growth rates for 59 bull sharks, Carcharhinus leucas, collected from the northern Gulf of Mexico were estimated from the band patterns formed seasonally in the vertebral centra. The combined age at length data for both sexes were applied to a von Bertalanffy growth model producing parameter estimates of L = 285 cm TL, K = .076, t₀ = –3.0 yr. Lengths at age for males and females were similar except that males did not attain as great a length as females. Growth was apparently slow and varied among individuals, but in general, was estimated to be 15–20 cm yr^–1 for the first five years, 10 cm yr^–1 for years 6–10, 5–7 cm yr^–1 for years 11–16, and less than 4–5 cm yr^–1 thereafter. Males mature at 210–220 cm TL or 14–15 yr of age; females mature at>225 cm TL or 18+ yr of age. The largest male (245 cm TL) was 21.3 yr old; the largest female (268 cm TL) was 24.2 yr old.