Multiple ciguatoxins present in Indian Ocean reef fish

Optimised gradient reversed-phase high-performance liquid chromatography electrospray ionisation mass spectrometry (LC/MS) methods, in combination with a [H-3]-brevetoxin binding assay (RLB), revealed multiple ciguatoxins in a partially purified extract of a highly toxic Lutjanus sebae (red emperor) from the Indian Ocean. Two major ciguatoxins of 1140.6 Da (I-CTX-1 and -2) and two minor ciguatoxins of 1156.6 Da (I-CTX-3 and -4) were identified. Accurate mass analysis revealed that I-CTX-1 and -2 and Caribbean C-CTX-1 had indistinguishable masses (1140.6316 Da, at 0.44 ppm resolution). Toxicity estimated from LC/MS/RLB responses indicated that I-CTX-1 and -2 were both similar to 60% the potency of Pacific ciguatoxin-1 (P-CTX-1). In contrast to ciguatoxins of the Pacific where the more oxidised ciguatoxins are more potent, I-CTX-3 and -4 were similar to 20% of P-CTX-1 potency. Interconversion in dilute acid or on storage, typical of spiroketal and hemiketal functionality found in P-CTXs and C-CTXs, respectively, was not observed to occur between I-CTX-1 and -2. The ratio of CTX-1 and -2 varied depending on the fish extract being analysed. These results suggest that I-CTX-1 and -2 may arise from separate dinoflagellate precursors that may be oxidatively biotransformed to I-CTX-3 and -4 in fish. (C) 2002 Published by Elsevier Science Ltd.

Effect of fertilizer placement on nitrogen loss from sugarcane in tropical Queensland

This paper reports on the fate of nitrogen (N) in a first ratoon sugarcane (Saccharum officinarum L.) crop in the wet tropics of Queensland when urea was either surface applied or drilled into the soil 3-4 days after harvesting the plant cane. Ammonia volatilization was measured with a micrometeorological method, and fertilizer N recovery in plants and soil, to a depth of 140 cm, was determined by mass balance in macroplots with N labelled urea 166 and 334 days after fertilizer application. The bulk of the fertilizer and soil N uptake by the sugarcane occurred between fertilizing and the first sampling on day 166. Nitrogen use efficiency measured as the recovery of labelled N in the plant was very low. At the time of the final sampling (day 334), the efficiencies for the surface and subsurface treatments were 18.9% and 28.8%, respectively. The tops, leaves, stalks and roots in the subsurface treatment contained significantly more fertilizer N than the corresponding parts in the surface treatment. The total recoveries of fertilizer N for the plant-trash-soil system on day 334 indicate significant losses of N in both treatments ( 59.1% and 45.6% of the applied N in the surface and subsurface treatments, respectively). Drilling the urea into the soil instead of applying it to the trash surface reduced ammonia loss from 37.3% to 5.5% of the applied N. Subtracting the data for ammonia loss from total loss suggests that losses by leaching and denitrification combined increased from 21.8% and 40.1% of the applied N as a result of the change in method of application. While the treatment resulted in increased denitrification and/or leaching loss, total N loss was reduced from 59.1% to 45.6%, ( a saving of 13.5% of the applied N), which resulted in an extra 9.9% of the applied N being assimilated by the crop.

The influence of increased line-fishing mortality on the sex ratio and age of sex reversal of the venus tusk fish

The age of sex reversal of the venus tusk fish Choerodon venustus, caught by line fishing at various locations on the southern Great Barrier Reef, indicated that C. venustus is capable of modifying its life cycle in response to increased mortality. The evidence suggests Masthead Reef fish, which experience the highest mortality, underwent sex reversal at a smaller size and younger age than at the other sites. The largest female fish, sexually transitional fish and males were smaller at Masthead Reef than at the Swains Reefs or One Tree Reef at Masthead Reef. There was also considerable overlap in the size of males and females within the exploited populations indicating that sex reversal is not initiated at a particular length but may have a social cause. The sex ratio of fish was essentially the same for fish fully susceptible to line fishing in the Swains and Masthead samples. Circumstantial evidence suggested that the absence of large males in a population may initiate sex reversal, indicating the maintenance of a constant sex ratio may have a social basis. (C) 2002 The Fisheries Society of the British Isles.

The effect of sodium chloride supplementation on the milk production of grazing Holstein Friesian cows during summer and autumn in a humid sub-tropical environment

Two studies were conducted to examine the effects of including NaCl at various rates in grain-based supplements for Friesian cows grazing established, dominant (>90%), rainfed kikuyu (Pennisetum clandestinum cv. Common) pastures during summer and autumn in a humid sub-tropical environment. In study 1 (19 January-27 March 1998), 48 cows (36 multiparous, 12 primiparous; 27-96 days postpartum) were allocated to one of four groups based on genetic merit, milk production, liveweight (LW) and days postpartum. They were fed (2.7 kg dry matter (DM) per cow, twice-a-day) one of four isoenergetic and isonitrogenous barley grain-based concentrates containing NaCl at concentrations (% as-fed) of either 0 (SC1), 1.1 (SC2), 2.2 (SC3) or 3.3 (SC4). Maximum temperature humidity index (THImax) was greater than or equal to78 during 50% of the experimental period. Concentrate NaCl content had no effect (P>0.05) on daily milk yield or LW change but daily yields of 4% fat corrected milk (FCM), fat and protein were higher (P0.05) among treatments at 7.6+/-1.24 kg DM per cow. In study 2 (18 January 1999-1 March 1999), 48 cows (32 pluriparous, 16 primiparous: 32-160 days postpartum) were fed (2.7 kg DM per cow twice-a-day) one of two isoenergetic and isonitrogenous barley grain-based concentrates containing NaCl at concentrations (% as-fed) of 0 (control) or 2.2 (HSC). THImax was greater than or equal to78 during 34% of days in the experimental period. Yields of milk, FCM, fat and protein were lower (P0.05) by concentrate NaCl content. These studies indicate that NaCl supplementation can be beneficial in terms of milk production during warm, humid conditions as opposed to milder conditions. (C) 2002 Elsevier Science B.V. All rights reserved.

Ross River virus disease in tropical Queensland: evolution of rheumatic manifestations in an inception cohort followed for six months

Objective: To describe the natural history of rheumatic manifestations of Ross River virus (RRV) disease. Design: Prospective longitudinal clinical review. Setting: North Queensland local government areas of Cairns, Douglas, Mareeba and Atherton during January to May 1998. Participants: General practice patients diagnosed with RRV disease on the basis of symptoms and a positive RRV IgM result. Main outcome measures: Rheumatic symptoms and signs assessed as soon as possible after disease onset and on two subsequent occasions (up to 6.5 months after onset). Results: 57 patients were recruited, 47 of whom were reviewed three times (at means of 1.1, 2.4 and 3.6 months after disease onset). Results are reported for these 47: 46 (98%) complained of joint pain at first review, with the ankles, wrists, fingers, knees and metacarpophalangeal joints (II-IV) most commonly involved. Prevalence of joint pain decreased progressively on second and third reviews, both overall (92% and 68% of patients, respectively), and in the five joints most commonly affected. The prevalence of other common rheumatic symptoms and signs, and use of non-steroidal anti-inflammatory drugs, also progressively declined over the three reviews. Conclusions: Earlier studies may have overestimated the prevalence and duration of symptoms in RRV disease. Progressive resolution over 3-6 months appears usual.

The effects of level of dietary protein on the milk production and rumen physiology of dairy cows fed a diet based on a tropical grass hay

zFour rumen-fistulated, multiparous Holstein-Friesian cows in early lactation were offered mixed diets based on rhodes grass hay (Chloris gayana) cv. Callide containing 13, 14, 15 or 16% crude protein (CP) on a dry matter basis, in a 4 x 4 latin square design. The estimated undegradable protein concentration in these diets was similar with rumen degradable protein concentration varying. Cows fed a diet containing 13% CP had lower (P = 0.07) milk yields than cows in other treatments (20.4 vs 21.9, 22.0 and 22.2 L/d for 13, 14, 15 and 16% CP, respectively). A positive linear relationship was found (P = 0.06) between organic matter intake and dietary CP%. There were negative linear relationships between dietary CP% and digestibilities of dry matter (P = 0.09), organic matter (P = 0.06) and neutral detergent fibre (P = 0.02). Feeding a diet containing 13% CP resulted in significantly lower (P = 0.001) molar proportions (%) of rumen valerate in comparison with other treatments. The molar proportions of isovalerate differed (P = 0.001) between treatments (0.66, 0.78, 0.89 and 1.04%) for 13, 14, 15 and 16% CP, respectively). Dietary protein level had no effect on rates of passage, in situ digestion of rhodes grass hay or ratios of allantoin: creatinine in urine. These data showed that increasing the dietary CP concentration of lactating cows fed diets based on rhodes grass hay increased intakes and not significantly improved at dietary CP concentrations above 14% DM.

Streptococcus pyogenes prtFII, but not sfbI, sfbII or fbp54, is represented more frequently among invasive-disease isolates of tropical Australia

Streptococcus pyogenes (group A streptococcus) strains may express several distinct fibronectin-binding proteins (FBPs) which are considered as major streptococcal adhesins. Of the FBPs, SfbI was shown in vitro to promote internalization of the bacterium into host cells and has been implicated in persistence. In the tropical Northern Territory, where group A streptococcal infection is common, multiple genotypes of the organism were found among isolates from invasive disease cases and no dominant strains were observed. To determine whether any FBPs is associated with invasive disease propensity of S. pyogenes, we have screened streptococcal isolates from bacteraemic and necrotizing fasciitis patients and isolates from uncomplicated infections for genetic endowment of 4 FBPs. No difference was observed in the distribution of sfbII, fbp54 and sfbI between the blood isolates' and isolates from uncomplicated infection. We conclude that the presence of sfbI does not appear to promote invasive diseases, despite its association with persistence. We also show a higher proportion of group A streptococcus strains isolated from invasive disease cases possess prtFII when compared to strains isolated from non-invasive disease cases. We suggest that S. pyogenes may recruit different FBPs for different purposes.

Analysis of toxin profiles in three different fish species causing ciguatera fish poisoning in Guadeloupe, French West Indies

A grey snapper (Lutjanus griseus), a grouper (Serranidae) and a blackjack (Caranx lugubris) were implicated in three different ciguatera poisonings in Guadeloupe, French West Indies. A mouse bioassay indicated toxicity for each specimens: 0.5-1, greater than or equal to 1 and > 1 M Ug g(-1), respectively. After purification by gel filtration chromatography, the samples were analysed by high-performance liquid chromatography coupled to mass spectrometry (LC-MS). The toxin profiles differ from one fish to another. C-CTX-1 was detected at 0.24, 0.90 and 13.8 ng g(-1) flesh in the snapper, grouper and jack, respectively. It contributed only to part of the whole toxicity determined by the mouse bioassay. Other toxins identified were C-CTX-2 (a C-CTX-1 epimer), three additional isomers of C-CTX-1 or -2, and five ciguatoxin congeners (C-CTX-1127, C-CTX-1143 and its isomer C-CTX-1143a, and C-CTX-1157 and its isomer C-CTX-1157b). Putative hydroxy-polyether-like compounds were also detected in the flesh of the grouper with [M+ + H](+) ions at m/z 851.51, 857.50, 875.51, 875.49 and 895.54 Da. Some of these compounds have the same mass range as some known dinoflagellate toxins. In conclusion, this study confirms the usefulness of LC-MS analysis to determine the ciguatoxins levels and the toxin profile in fish flesh hazardous to humans.

Independent and conjugate eye movements during optokinesis in teleost fish

In response to movements involving a large part of the visual field, the eyes of vertebrates typically show an optokinetic nystagmus, a response in which both eyes are tightly yoked. Using a comparative approach, this study sets out to establish whether fish with independent spontaneous eye movements show independent optokinetic nystagmus in each eye. Two fish with independent spontaneous eye movements, the pipefish Corythoichthyes intestinalis and the sandlance Limnichthyes fasciatus were compared with the butterflyfish Chaetodon rainfordi, which exhibits tightly yoked eye movements. In the butterflyfish a single whole-field stimulus elicits conjugate optokinesis, whereas the sandlance and pipefish show asynchronous optokinetic movements. In a split drum experiment, when both eyes were stimulated in opposite directions with different speeds, both the sandlance and the pipefish compensated independently with each eye. The optokinetic response in the butterflyfish showed some disconjugacy but was generally confused. When one eye was occluded, the seeing eye was capable of driving the occluded eye in both the butterflyfish and the pipefish but not in the sandlance. Monocular occlusion therefore unmasks a link between the two eyes in the pipefish, which is overridden when both eyes receive visual input. The sandlance never showed any correlation between the eyes during optokinesis in all stimulus conditions. This suggests that there are different levels of linkage between the two eyes in the oculomotor system of teleosts, depending on the visual input.

delta N-15 values of tropical savanna and monsoon forest species reflect root specialisations and soil nitrogen status

A large number of herbaceous and woody plants from tropical woodland, savanna, and monsoon forest were analysed to determine the impact of environmental factors (nutrient and water availability, fire) and biological factors (microbial associations, systematics) on plant delta(15)N values. Foliar delta(15)N values of herbaceous and woody species were not related to growth form or phenology, but a strong relationship existed between mycorrhizal status and plant delta(15)N. In woodland and savanna, woody species with ectomycorrhizal (ECM) associations and putative N-2-fixing species with ECM/arbuscular (AM) associations had lowest foliar delta(15)N values (1.0-0.6parts per thousand), AM species had mostly intermediate delta(15)N values (average +0.6parts per thousand), while non-mycorrhizal Proteaceae had highest delta(15)N values (+2.9 to +4.1parts per thousand). Similar differences in foliar delta(15)N were observed between AM (average 0.1 and 0.2parts per thousand) and non-mycorrhizal (average +0.8 and +0.3parts per thousand) herbaceous species in woodland and savanna. Leguminous savanna species had significantly higher leaf N contents (1.8-2.5% N) than non-fixing species (0.9-1.2% N) indicating substantial N acquisition via N-2 fixation. Monsoon forest species had similar leaf N contents (average 2.4% N) and positive delta(15)N values (+0.9 to +2.4parts per thousand). Soil nitrification and plant NO3- use was substantially higher in monsoon forest than in woodland or savanna. In the studied communities, higher soil N content and nitrification rates were associated with more positive soil delta(15)N and plant delta(15)N. In support of this notion, Ficus, a high NO3- using taxa associated with NO3- rich sites in the savanna, had the highest delta(15)N values of all AM species in the savanna. delta(15)N of xylem sap was examined as a tool for studying plant delta(15)N relations. delta(15)N of xylem sap varied seasonally and between differently aged Acacia and other savanna species. Plants from annually burnt savanna had significantly higher delta(15)N values compared to plants from less frequently burnt savanna, suggesting that foliar N-15 natural abundance could be used as marker for assessing historic fire regimes. Australian woodland and savanna species had low leaf delta(15)N and N content compared to species from equivalent African communities indicating that Australian biota are the more N depauperate. The largest differences in leaf delta(15)N occurred between the dominant ECM Australian and African savanna (miombo) species, which were depleted and enriched in N-15, respectively. While the depleted delta(15)N of Australian ECM species are similar to those of previous reports on ECM species in natural plant communities, the N-15-enriched delta(15)N of African ECM species represent an anomaly.

Adaptation of rainbow fish to lake and stream habitats

Fish occupy a range of hydrological habitats that exert different demands on locomotor performance. We examined replicate natural populations of the rainbow fishes Melanotaenia eachamensis and M. duboulayi to determine if colonization of low-velocity (lake) habitats by fish from high-velocity (stream) habitats resulted in adaptation of locomotor morphology and performance. Relative to stream conspecifics, lake fish had more posteriorly positioned first dorsal and pelvic fins, and shorter second dorsal fin bases. Habitat dimorphism observed between wild-caught fish was determined to be heritable as it was retained in M. eachamensis offspring raised in a common garden. Repeated evolution of the same heritable phenotype in independently derived populations indicated body shape divergence was a consequence of natural selection. Morphological divergence between hydrological habitats did not support a priori expectations of deeper bodies and caudal peduncles in lake fish. However, observed divergence in fin positioning was consistent with a family-wide association between habitat and morphology, and with empirical studies on other fish species. As predicted, decreased demand for sustained swimming in takes resulted in a reduction in caudal red muscle area of lake fish relative to their stream counterparts. Melanotaenia duboulayi lake fish also had slower sustained swimming speeds (U-crit) than stream conspecifics. In M. eachamensis, habitat affected U-crit of males and females differently. Specifically, females exhibited the pattern observed in M. duboulayi (lake fish had faster U-crit than stream fish), but the opposite association was observed in males (stream males had slower Ucrit than lake males). Stream M. eachamensis also exhibited a reversed pattern of sexual dimorphism in U-crit (males slower than females) relative to all other groups (males faster than females). We suggest that M. eachamensis males from streams responded to factors other than water velocity. Although replication of muscle and U,,it phenotypes across same habitat populations within and/or among species was suggestive of adaptation, the common garden experiment did not confirm a genetic basis to these associations. Kinematic studies should consider the effect of the position and base length of dorsal fins.

Behavioural responses of two native Australian fish species (Melanotaenia duboulayi and Pseudomugil signifer) to introduced poeciliids (Gambusia holbrooki and Xiphophorus helleri) in controlled conditions

Experimental treatments to compare behavioural responses included native fish species only, natives plus one exotic species and natives plus both exotic species. The mosquitofish, Gambusia holbrooki frequently attacked both native species, but tended to nip Melanotaenia duboulayi (especially small individuals) and chase Pseudomugil signifer The frequency of attacks by G. holbrooki on M. duboulayi rose when all four fish species were present. When food was added, all four species showed a strong increase in aggression, especially in the four-species treatment, where there were significant increases in the frequency of attacks by the swordtail Xiphophorus helleri on M. duboulay and by M. duboulayi on G. holbrooki, and of conspecific attacks by M. duboulayi. Increased attack frequency was associated with aggregation closer to the water's surface, regardless of the presence of food. The results support the hypothesis that introduced poeciliids can have deleterious competitive effects on native species. However, while juvenile M. duboulayi were particularly vulnerable to the secondary, effects of fin-nipping, R signifer appeared to be more susceptible to physical displacement and reduced food capture success.

Cleaner fish drives local fish diversity on coral reefs

Coral reefs are one of the most diverse habitats in the world [1], yet our understanding of the processes affecting their biodiversity is limited [1-3]. At the local scale, cleaner fish are thought to have a disproportionate effect, in relation to their abundance and size, on the activity of many other fish species, but confirmation of this species' effect on local fish diversity has proved elusive. The cleaner fish Labroides dimidiatus has major effects on fish activity patterns [4] and may indirectly affect fish demography through the removal of large numbers of parasites [5, 6]. Here we show that small reefs where L. dimidiatus had been experimentally excluded for 18 months had half the species diversity of fish and one-fourth the abundance of individuals. Only fish that move among reefs, however, were affected. These fish include large species that themselves can affect other reef organisms [2, 7]. In contrast, the distribution of resident fish was not affected by cleaner fish. Thus, many fish appear to choose reefs based on the presence of cleaner fish. Our findings indicate that a single small [8] and not very abundant [9] fish has a strong influence on the movement patterns, habitat choice, activity, and local diversity and abundance of a wide variety of reef fish species.