Larval biomass and chemical composition at hatching in two geographically isolated clades of the shrimp Macrobrachium amazonicum: intra- or interspecific variation?

The shrimp Macrobrachium amazonicum (Heller 1862) has an extremely large geographic range (>4000?km across) in northern and central South America, comprising estuarine and fully limnic inland populations, which are hydrologically isolated from each other. Significant variations in ecology, physiology, reproduction, and larval development suggest an at least incipient allopatric speciation due to limited genetic exchange. In a comparative experimental investigation with shrimps from the Pantanal (upper Paraguay River basin) and the Amazon delta, respectively, we measured larval body size, dry weight (W), biochemical (total protein; lipid; fatty acids, FA), and elemental composition (carbon, hydrogen, nitrogen; collectively CHN) at hatching. All these early larval traits are relevant for the degree of developmental dependence on planktonic food sources. Various consistent differences were observed between the two populations: Newly hatched larvae produced by shrimps from the Amazon delta were significantly smaller and showed lower values of W, CHN, protein, and unsaturated FA compared to those from the Pantanal. On the other hand, they contained significantly higher quantities of total lipid and saturated FA and, in consequence, higher ratios of lipid:protein, C:N, and saturated:unsaturated FA. All these differences in biomass and chemical composition suggest that the larvae of the Amazon population are energetically better adapted to planktonic food limitation, which likely occurs during riverine downstream transport toward coastal marine waters, also explaining previous observations of much stronger initial starvation tolerance in larvae from the Amazon versus those from the Pantanal. The latter develop in highly productive lentic inland waters, where large body size, an early onset of feeding, and a strong musculature (indicated by a high protein content) should facilitate their role as planktonic predators and allow for fast growth. An initial independence from food (lecithotrophy in the zoea I stage) as well as a preference for oligohaline rather than fully limnic conditions observed in the Pantanal larvae are interpreted as traits that have persisted from an ancestral coastal marine clade. Altogether, consistent ontogenetic differences between shrimps from the Pantanal and the Amazon estuary support the hypothesis that the taxon M. amazonicum comprises a complex of closely related but separate species.     

A study on the energy source in the developing embryo of the mangrove horseshoe crab,Carcinoscorpius rotundicauda (Latreille)

The mangrove horseshoe crab, Carcinoscorpius rotundicauda, is an Asian species found in Malaysia. This ancient arthropod has a long incubational period during which it depends on various energy sources for both embryogenesis and organogenesis. This study describes the trend of energy utilization from the endogenous reserves by the developing embryos from 0 to 40 days of incubation (until the hatching of the larvae). The dry weight, insoluble protein, carbohydrate, glycogen and lipid showed a declining trend from 0 to 40 days of incubation, whereas the wet weight, water content, ash content and soluble protein showed an increasing trend. Selected micro-elements such as Cu²⁺, Fe²⁺ and Zn²⁺ also demonstrated an interesting trend in the developing eggs when the egg mass was subjected to inductively coupled plasma mass spectrometry analysis, where these elements showed a high correlation with the moulting stages and egg development. Maximum variations within the micro-elements were observed during the 1st (10 days after fertilization) and 2nd (35–36 days after fertilization) moulting stages within the developing eggs. This study clearly indicated that the moulting cycles of C. rotundicauda during embryonic development influence energy utilization in the form of carbohydrates, lipids, proteins, glycogen and other micro-elements.     

The effect of salinity on larval survival and development in the sea urchin Echinometra lucunter

Summary

Reductions in salinity can have adverse effects on larval development and larval survival in some invertebrate taxa but not others. Salinity tolerance of larvae may be particularly important in echinoderms because they are both poor ion regulators and stenohaline. I examined the effect of six levels of salinity (15, 18, 21, 24, 27 and 33 PSU) on survival and rate of development of larvae in the subtropical sea urchin Echinometra lucunter. In the short-term, mortality rate was significantly lower in 33 PSU than in all other salinities except 27 PSU, and it was significantly greater in 15 and 18 PSU than in all higher salinities. In the long-term, daily and cumulative mortality were significantly greater in 15 PSU than in most other salinities over 11 days of development (except for cumulative mortality in 18 PSU). They were significantly greater in 18 PSU than in 21 PSU or 33 PSU over a period of 13 days. Furthermore, daily mortality was significantly greater in 18 PSU than in 24 PSU or 27 PSU at 13 d after fertilization. Daily and cumulative mortality were significantly lower in 33 PSU than in 21, 24 or 27 PSU over a period of 17 days. Although in the control (33 PSU) 75% of larvae completed development to the 8-arm stage at 35 d, no larvae developed further than the 4-arm stage in 18, 21, 24 or 27 PSU; in 15 PSU, ~60% of larvae did not develop further than swimming blastulae. Since prolonged exposure to salinities as high as 27 PSU (frequently recorded in the adult habitat) can result in great larval losses, adaptive behaviours that prevent larvae from entering water layers of low salinity will enhance their chance for survival.     

Mini review: Hydrodynamics of larval settlement into fouling communities

Abstract

Flowing water delivers planktonic larvae to surfaces, but also dislodges them. This paper reviews experiments in the field and in laboratory flumes, as well as mathematical models, which have revealed how the interaction of ambient water motion with a developing fouling community affects larval settlement. Although mean current velocities across fouling communities in harbours are low, instantaneous velocities can be much higher due to turbulence and to the velocity oscillations of wind chop and ship wakes. As a fouling community develops, its topography becomes more complex and the range of flow microhabitats on the spatial scale of larvae increases. In spite of the prevalence of waves in shallow coastal habitats, and in spite of the importance to settlement of the fine-scale instantaneous velocities encountered by larvae, most studies of flow effects on larval settlement have focused on unidirectional currents and on temporally- and spatially-averaged aspects of the flow.     

Oxygen-depleted surfaces: a new antifouling technology

A novel, non-toxic strategy to combat marine biofouling is presented. The technology is paint with additions of up to 43% of industrial protein. Through microbial degradation of the protein component, an oxygen-depleted layer rapidly forms in a 0.2 mm layer close to the paint surface. With the present paint formulations, a stable, O₂-depleted layer can persist for 16 weeks. Barnacle larvae (cyprids) did not settle on panels where oxygen saturation was <20%, and cyprids were killed when exposed to O₂-free water for more than 1 h. It is also shown that the O₂-depleted layer will rapidly reform (within 15 min) after exposure to turbulent flow. Field exposure of panels for 16 weeks showed that paint with protein reduced fouling by barnacles and bryozoans by 80% and close to 100%, respectively. The results suggest that this novel technology may be developed into a non-toxic alternative to copper-based antifouling paints, especially for pleasure boats in sensitive environments. There is clearly potential for further development of the paint formulation, and a full-scale test on a boat-hull suggested that service-life under realistic operations needs to be improved.     

食品级惰性粉对三种储藏物害虫生长发育的影响

应用粉剂药膜法,一种食品级惰性粉——4号粉相应剂量处理赤拟谷盗Tribolium castaneum(Herbst)、烟草甲Lasioderma serricorne(Fabricius)、锯谷盗Oryzaephilus surinamensis(Linnaeus)的卵和1龄幼虫,赤拟谷盗、锯谷盗、烟草甲的卵平均孵化率均在97%以上,处理后卵孵化的1龄幼虫平均死亡率均在96%以上;处理3种害虫的1龄幼虫其平均死亡率均在99%以上。结果表明食品级惰性粉对3种试虫卵的孵化率几乎没有影响,但对其1龄幼虫有较好的防治效果。通过混粮法应用正交试验研究发现,惰性粉剂量和小麦含水量对赤拟谷盗F1代防治效果均有显著影响。当小麦处于安全水分时(含水量12%¹4%),10014%),100¹50 mg/kg 4号粉能有效防治赤拟谷盗F1代。     

Settlement Behaviour of Marine Invertebrate Larvae Measured by EthoVision 3.0

Submerged marine surfaces are rapidly colonized by fouling organisms. Current research is aimed at finding new, non-toxic, or at least environmentally benign, solutions to this problem. Barnacles are a major target organism for such control as they constitute a key component of the hard fouling community. A range of standard settlement assays is available for screening test compounds against barnacle cypris larvae, but they generally provide little information on mechanism(s) of action. Towards this end, a quick and reliable video-tracking protocol has been developed to study the behaviour of the cypris larvae of the barnacle, Balanus amphitrite, at settlement. EthoVision 3.0 was used to track individual cyprids in 30-mm Petri dishes. Experiments were run to determine the optimal conditions vis-à-vis acclimation time, tracking duration, number of replicates, temperature and lighting. A protocol was arrived at involving a two Petri dish system with backlighting, and tracking over a 5-min period after first acclimating the cyprids to test conditions for 2 min. A minimum of twenty replicates was required to account for individual variability in cyprid behaviour from the same batch of larvae. This methodology should be widely applicable to both fundamental and applied studies of larval settlement and with further refinements, to that of smaller fouling organisms such as microalgae and bacteria.     

Lethal and sub‐lethal impacts of pulsed laser irradiations on the larvae of the fouling barnacle Balanus amphitrite

Laboratory experiments were conducted to study the impact of laser irradiation on the larvae of the fouling barnacle Balanus amphitrite. Research pertaining to fouling invertebrate larvae‐laser interaction is sparse and, hence, data on this aspect were thought significant in order to consider pulsed low power laser irradiations as a possible future antifouling tool. Lethal and sub‐lethal impacts of four very low laser fluences, viz. 0.013, 0.025, 0.05 and 0.1 J cm^‐2 for three different durations, viz. 2, 10 and 30 s were investigated. Three growth stages of barnacle larvae, viz. nauplii stage II, nauplii stage IV and cyprids were exposed to the mentioned laser fluences for different durations. While lethal impact was assessed immediately after and 1 d after irradiation, sub‐lethal impacts were studied by monitoring the success rate of the irradiated nauplii in reaching the cyprid stage. In addition, the swimming speed of VI^th stage nauplii after irradiation was studied. In the case of cyprids, in addition to the mortality measurement immediately after and 1 d after irradiation, the settlement rate was investigated. In all the above experiments, non‐irradiated larvae served as controls. The results showed an increase in mortality with increasing laser fluence and duration of irradiation. Irradiation for 2 s resulted in significant mortality in nauplii, while it was less in the case of cyprids. In II^nd stage nauplii, the mortality immediately after irradiation for 2 s varied from 14.8±2.12 to 97.1±4.1% for laser fluences of 0.013 and 0.1 J cm^‐2, respectively. However, in cyprids, the mortality immediately after irradiation for 2 s varied from 12.2±3 to 13.4±1.2% for fluences of 0.013 and 0.1 J cm^‐2, respectively. The mortality in IV^th stage nauplii was less than that for II^nd stage nauplii but more than that for cyprids. There was a significant increase in mortality with time after irradiation. The formation of cyprids from the irradiated larvae was significantly less than that observed for non‐irradiated larvae. Also, the irradiated larvae showed a significantly slower swimming speed compared to the control samples. The settlement rate in cyprids was reduced significantly by the laser irradiation. This was true even for the lowest fluence and shortest period of irradiation tested. Thus, the results of the experiment showed that even a low power pulsed laser irradiation of 0.013 J cm^‐2 for 2 s can cause significant damage to fouling barnacle larvae.     

Ontogeny of body density and the swimbladder in yellowtail kingfish Seriola lalandi larvae

The ontogeny of larval body density and the morphological and histological events during swimbladder development were investigated in two cohorts of yellowtail kingfish Seriola lalandi larvae to understand the relationship between larval morphology and body density. Larvae <3 days post hatch (dph) were positively buoyant with a mean ± s.d . body density of 1·023 ± 0·001 g cm^?3. Histological evidence demonstrated that S. lalandi larvae are initially transient physostomes with the primordial swimbladder derived from the evagination of the gut ventral to the notochord and seen at 2 dph. A pneumatic duct connected the swimbladder to the oesophagus, but degenerated after 5 dph. Initial swimbladder (SB) inflation occurred on 3 dph, and the inflation window was 3–5 dph when the pneumatic duct was still connected to the gut. The swimbladder volume increased with larval age and the epithelial lining on the swimbladder became flattened squamous cells after initial inflation. Seriola lalandi developed into a physoclist with the formation of the rete mirabile and the gas‐secreting gland comprised low‐columnar epithelial cells. Larvae with successfully inflated swimbladders remained positively buoyant, whereas larvae without SB inflation became negatively buoyant and their body density gradually reached 1·030 ± 0·001 g cm^?3 by 10 dph. Diel density changes were observed after 5 dph, owing to day time deflation and night‐time inflation of the swimbladder. These results show that SB inflation has a direct effect on body density in larval S. lalandi and environmental factors should be further investigated to enhance the rate of SB inflation to prevent the sinking death syndrome in the early life stage of the fish larvae.     

Brugia pahangi: Immunization with early L3 ES alters parasite migration,and reduces microfilaremia and lymphatic lesion formation in gerbils (Meriones unguiculatus)

Previous studies have shown that intradermally (ID) injected Brugia pahangi L3s migrate through various tissues and into the lymphatics of gerbils in a distinct pattern. Excretory/secretory products (ES) produced at the time of invasion of B. pahangi are likely to be important in this early migration phase of the parasite life cycle in their rodent host. Hence, early L3 ES was collected from 24 h in vitro cultures of B. pahangi L3 larvae and used in immunization experiments to investigate the effect of immunity to early L3 ES on worm migration, survival and development of B. pahangi. Immunization of gerbils with ES in RIBI adjuvant produced antibodies to numerous ES proteins eliciting a strong humoral response to ES and indirect fluorescent antibody (IFA) assay using anti-ES serum recognized the ES proteins on the surface of B. pahangi L3 larvae. Following ES immunization, gerbils were challenged either ID or intraperitoneally (IP) with 100 L3s of B. pahangi and euthanized at 3 or 106 days post inoculation (DPI). Immunization with early ES slowed the migration of ID inoculated L3 at 3 DPI and significantly altered the locations of adult worms at 106 DPI. Immunization did not induce protection in any treatment group. However, immunized animals had significantly fewer microfilariae per female worm suggesting the antigens in ES are important in microfilariae development or survival in the host. The number of lymphatic granulomas was also significantly reduced in ES immunized animals. It is important to note that microfilariae serve as a nidus in these granulomas. Our results shows immunization with early Brugia malayi L3 ES alters the worm migration, affects circulating microfilarial numbers and reduces lymphatic granulomas associated with B. pahangi infection in gerbils.