A synthesis of dominant ecological processes in intensive shrimp ponds and adjacent coastal environments in NE Australia

One of the key environmental concerns about shrimp farming is the discharge of waters with high levels of nutrients and suspended solids into adjacent waterways. In this paper we synthesize the results of our multidisciplinary research linking ecological processes in intensive shrimp ponds with their downstream impacts in tidal, mangrove-lined creeks. The incorporation of process measurements and bioindicators, in addition to water quality measurements, improved our understanding of the effect of shrimp farm discharges on the ecological health of the receiving water bodies. Changes in water quality parameters were an oversimplification of the ecological effects of water discharges, and use of key measures including primary production rates, phytoplankton responses to nutrients, community shifts in zooplankton and delta(15)N ratios in marine plants have the potential to provide more integrated and robust measures. Ultimately, reduction in nutrient discharges is most likely to ensure the future sustainability of the industry. (C) 2003 Elsevier Ltd. All rights reserved.

Enrichment of nitrifying microbial communities from shrimp farms and commercial inocula

Nitrifying bacteria were selected from shrimp farm water and sediment (natural seed) in Thailand and from commercial seed cultures. The microbial consortia from each source giving the best ammonia removal during batch culture pre-enrichments were used as inocula for two sequencing batch reactors (SBRs). Nitrifiers were cultivated in the SBRs with 100 mg NH4-N/I and artificial wastewater containing 25 ppt salinity. The two SBRs were operated at a 7 d hydraulic retention time (HRT) for 77 d after which the HRT was reduced to 3.5 d. The amounts of ammonia removed from the influent by microorganisms sourced from the natural seed were 85% and 92% for the 7 d HIRT and the 3.5 d HRT, respectively. The ammonia removals of microbial consortia from the commercial seed were 71% and 83% for these HRTs respectively. The quantity of ammonia-oxidizing bacteria (AOB) and nitrite-oxidizing bacteria (NOB) was determined in the SBRs using the most probable number (MPN) technique. Both AOB and NOB increased in number over the long-term operation of both SBRs. According to quantitative fluorescence in situ hybridisation (FISH) probing, AOB from the natural seed and commercial seed comprised 21 +/- 2% and 30 +/- 2%, respectively of all bacteria. NOB could not be detected with currently-reported FISH probes, suggesting that novel NOB were enriched from both sources. Taken collectively, the results from this study provide an indication that the nitrifiers from shrimp farm sources are more effective at ammonia removal than those from commercial seed cultures.

Sea anemone host selection by the symbiotic saddled cleaner shrimp Periclimenes holthuisi

A preliminary field survey was conducted to determine the distribution of ectosymbiotic shrimp Periclimenes holthuisi on the sea anemone Stichodactyla haddoni in Moreton Bay (Queensland, Australia). Laboratory experiments were also carried out to verify whether the shrimp show a preference for one anemone host. In the field, 45 individuals of P. holthuisi were found to be associated with 70% of the specimens of S. haddoni (n=20). We inferred this shrimp population was not space-limited because not all anemones were colonized. After having been isolated from their natural host for 2 weeks, when placed between individuals of S. haddoni and Macrodactyla doreensis (an anemone that is sympatric with S. haddoni), shrimp overwhelmingly selected S. haddoni (92%). To establish whether M. doreensis may serve as an alternative host for P. holthuisi, unacclimated shrimp were forced to associate with this anemone. Macrodactyla doreensis showed little tentacle reaction during this association; shrimp were found on the anemone's tentacles and the column. The finding that M. doreensis can serve as an alternative host for P. holthuisi demonstrates that this anemoneshrimp is adaptable to another anemone host and thus may not be highly host specific.

Assessing the influence and distribution of shrimp pond effluent in a tidal mangrove creek in north-east Australia

Effluent from a land based shrimp farm was detected in a receiving creek as changes in physical, chemical and biological parameters. The extent and severity of these changes depended on farm operations. This assessment was conducted at three different stages of shrimp-pond maturity, including (1) when the ponds were empty, (2) full and (3) being harvested. Methods for assessing farm effluent in receiving waters included physical/chemical analyses of the water column, phytoplankton bioassays and nitrogen isotope signatures of marine flora. Comparisons were made with an adjacent creek that served as the farms intake creek and did not directly receive effluent. Physical/chemical parameters identified distinct changes in the receiving creek with respect to farm operations. Elevated water column NH4+ (18.5+/-8.0 muM) and chlorophyll a concentrations (5.5+/-1.9 mug/l) were measured when the farm was in operation, in contrast to when the farm was inactive (1.3+/-0.3 muM and 1.2+/-0.6 mug/l, respectively). At all times, physically chemical parameters at the mouth of the effluent creek, were equivalent to control values, indicating effluent was contained within the effluent-receiving creek. However, elevated delta(15)N signatures of mangroves (up to similar to8parts per thousand) and macroalgae (up to similar to5parts per thousand) indicated a broader influence of shrimp farm effluent, extending to the lower regions of the farms intake creek. Bioassays at upstream sites close to the location of farm effluent discharge indicated that phytoplankton at these sites did not respond to further nutrient additions, however downstream sites showed large growth responses. This suggested that further nutrient loading from the shrimp farm, resulting in greater nutrient dispersal, will increase the extent of phytoplankton blooms downstream from the site of effluent discharge. When shrimp ponds were empty water quality in the effluent and intake creeks was comparable. This indicated that observed elevated nutrient and phytoplankton concentrations were directly attributable to farm operations. (C) 2003 Elsevier Ltd. All rights reserved.

Cleaner shrimp do clean

Apart from cleaner fish, there are many reports on cleaning by shrimps, yet whether shrimps actually 'clean', i.e. eat parasites in the wild, has not been demonstrated. For the first time, we show that, conclusively, cleaner shrimp in the wild do clean. We found crustacean ectoparasites from the Family Gnathiidae and the Class Copepoda in the gut contents of wild cleaner shrimp, Urocaridella sp. and Periclimenes holthuisi. In addition, they ate parasitic monogenean flatworms, Benedenia sp., offered to them in the laboratory. Finally, P. holthuisi, significantly reduced monogenean, Benedenia sp., loads by 74.5% on captive surgeonfish Ctenochaetus striatus within 48 h. Such large reductions in parasite loads are likely to benefit individual fish. These results emphasise the need for more information on the ecological role of cleaner shrimp on coral reefs.

Cleaner shrimp use a to advertise cleaning rocking dance service to clients

Signals transmit information to receivers about sender attributes, increase the fitness of both parties, and are selected for in cooperative interactions between species to reduce conflict [1, 2]. Marine cleaning interactions are known for stereotyped behaviors [3-6] that likely serve as signals. For example, dancing and tactile dancing in cleaner fish may serve to advertise cleaning services to client fish [7] and manipulate client behavior [8], respectively. Cleaner shrimp clean fish [9], yet are cryptic in comparison to cleaner fish. Signals, therefore, are likely essential for cleaner shrimp to attract clients. Here, we show that the yellow-beaked cleaner shrimp [110] Urocaridella sp. c [11] uses a stereotypical side-to-side movement, or rocking dance, while approaching potential client fish in the water column. This dance was followed by a cleaning interaction with the client 100% of the time. Hungry cleaner shrimp, which are more willing to clean than satiated ones [12], spent more time rocking and in closer proximity to clients Cephaiopholis cyanostigma than satiated ones, and when given a choice, clients preferred hungry, rocking shrimp. The rocking dance therefore influenced client behavior and, thus, appears to function as a signal to advertise the presence of cleaner shrimp to potential clients.

The potential of microarrays to assist shrimp breeding and production: a review

The shrimp aquaculture industry is a relatively new livestock industry, having developed over the past 30 years. Thus, it is poised to take advantage of new technologies from the outset of selective breeding programs. This contrasts with long established livestock industries, where there are already highly specialised breeds. This review focuses specifically on the potential application of microarrays to shrimp breeding. Potential applications of microarrays in selective breeding programs are summarised. Microarrays can be used as a rapid means to generate molecular markers for genetic linkage mapping, and genetic maps have been constructed for yeast, Arabidopsis and barley using microarray technology. Microarrays can also be used in the hunt for candidate genes affecting particular traits, leading to development of perfect markers for these traits (i.e. causative mutations). However, this requires that microarray analysis be combined with genetic linkage mapping, and that substantial genomic information is available for the species in question. A novel application of microarrays is to treat gene expression as a quantitative trait in itself and to combine this with linkage mapping to identify quantitative trait loci controlling the levels of gene expression; this approach may identify higher level regulatory genes in specific pathways. Finally, patterns of gene expression observed using microarrays may themselves be treated as phenotypic traits in selection programs (e.g. a particular pattern of gene expression might be indicative of a disease tolerant individual). Microarrays are now being developed for a number of shrimp species in laboratories around the world, primarily with a focus on identifying genes involved in the immune response. However, at present, there is no central repository of shrimp genomic information, which limits the rate at which shrimp genomic research can be progressed. The application of microarrays to shrimp breeding will be extremely limited until there is a shared repository of genomic information for shrimp, and the collective will and resources to develop comprehensive genomic tools for shrimp.

Design, development, and use of molecular primers and probes for the detection of Gluconacetobacter species in the pink sugarcane mealybug

Molecular tools for the species-specific detection of Gluconacetobacter sacchari, Gluconacetobacter diazotrophicus, and Gluconacetobacter liquefaciens from the pink sugarcane mealybug (PSMB) Saccharicoccus sacchari Cockerell (Homiptera: Pseudococcidae) were developed and used in polymerase chain reactions (PCR) and in fluorescence in situ hybridizations (FISH) to better understand the microbial diversity and the numerical significance of the acetic acid bacteria in the PSMB microenvironment. The presence of these species in the PSMB occurred over a wide range of sites, but not in all sites in sugarcane-growing areas of Queensland, Australia, and was variable over time. Molecular probes for use in FISH were also designed for the three acetic acid bacterial species, and shown to be specific only for the target species. Use of these probes in FISH of squashed whole mealybugs indicated that these acetic acid bacteria species represent only a small proportion of the microbial population of the PSMB. Despite the detection of Glac. sacchari, Glac. diazotrophicus, and Glac. liquefaciens by PCR from different mealybugs isolated at various times and from various sugarcane-growing areas in Queensland, Australia, these bacteria do not appear to be significant commensals in the PSMB environment.

Photoreceptor projection and termination pattern in the lamina of gonodactyloid stomatopods (mantis shrimp)

The apposition compound eyes of gonodactyloid stomatopods are divided into a ventral and a dorsal hemisphere by six equatorial rows of enlarged ommatidia, the mid-band (MB). Whereas the hemispheres are specialized for spatial vision, the MB consists of four dorsal rows of ommatidia specialized for colour vision and two ventral rows specialized for polarization vision. The eight retinula cell axons (RCAs) from each ommatidium project retinotopically onto one corresponding lamina cartridge, so that the three retinal data streams (spatial, colour and polarization) remain anatomically separated. This study investigates whether the retinal specializations are reflected in differences in the RCA arrangement within the corresponding lamina cartridges. We have found that, in all three eye regions, the seven short visual fibres (svfs) formed by retinula cells 1-7 (R1-R7) terminate at two distinct lamina levels, geometrically separating the terminals of photoreceptors sensitive to either orthogonal e-vector directions or different wavelengths of light. This arrangement is required for the establishment of spectral and polarization opponency mechanisms. The long visual fibres (lvfs) of the eighth retinula cells (R8) pass through the lamina and project retinotopically to the distal medulla externa. Differences between the three eye regions exist in the packing of svf terminals and in the branching patterns of the lvfs within the lamina. We hypothesize that the R8 cells of MB rows 1-4 are incorporated into the colour vision system formed by R1-R7, whereas the R8 cells of MB rows 5 and 6 form a separate neural channel from R1 to R7 for polarization processing.

Molecular investigation of the microbial populations of the pink sugarcane mealybug, Saccharicoccus sacchari

In an attempt to better understand the microbial diversity and endosymbiotic microbiota of the pink sugarcane mealybug (PSMB) Saccharicoccus sacchari Cockerell (Homoptera: Pseudococcidae), culture-independent approaches, namely PCR, a 16S rDNA clone library, and temperature gradient gel electrophoresis (TGGE) were used. Previous work has indicated that the acetic acid bacteria Gluconacetobacter sacchari, Gluconacetobacter diazotrophicus, and Gluconacetobacter liquefaciens represent only a small proportion of the microbial community of the PSMB. These findings were supported in this study by TGGE, where no bands representing G. sacchari, G. diazotrophicus, and G. liquefaciens on the acrylamide gel could be observed following electrophoresis, and by a 16S rDNA clone library study, where no clones with the sequence of an acetic acid bacterium were found. Instead, TGGE revealed that the mealybug microbial community was dominated by beta- and gamma-Proteobacteria. The dominant band in TGGE gels found in a majority of the mealybug samples was most similar, according to BLAST analysis, to the beta-symbiont of the craw mealybug Antonina crawii and to Candidatus Tremblaya princeps, an endosymbiont from the mealybug Paracoccus nothofagicola. The sequences of other dominant bands were identified as gamma-Proteobacteria, and were most closely related to uncultured bacterial clones obtained from soil samples. Mealybugs collected from different areas in Queensland, Australia, were found to produce similar TGGE profiles, although there were a few exceptions. A 16S rDNA clone library based on DNA extracted from a mealybug collected from sugarcane in the Burdekin region in Queensland, Australia, indicated very low levels of diversity among mealybug microbial populations. All sequenced clones were most closely related to the same members of the gamma-Proteobacteria, according to BLAST analysis.

Client fish ectoparasite loads and cleaner shrimp Urocaridella sp c hunger levels affect cleaning behaviour

Cleaning is a classic example of mutualism and determining the factors that maintain the balance between the costs and benefits for mutualist partners can assist our understanding of how cleaning relationships are maintained. Optimal foraging theory suggests two factors that might help to maintain the relationship between cleaners and their clients: client ectoparasite load and cleaner hunger levels. The ecological relevance and importance of foraging by cleaner fish in marine systems has been demonstrated repeatedly, yet there is little information available on this behaviour in cleaner shrimp. To determine whether cleaner shrimp base their choice of client fish on food patch quality (i.e. client fish ectoparasite load) we offered the yellow-beaked cleaner shrimp Urocaridella sp. c a choice of parasitized and unparasitized rock cods, Cephalopholis cyanostigma. To determine whether cleaner shrimp hunger levels influence cleaning time, we manipulated hunger levels in Urocaridella sp. c and examined their behaviour towards parasitized client fish. Cleaner shrimp preferred parasitized to unparasitized client fish and food-deprived cleaner shrimp cleaned parasitized rock cods more frequently than satiated cleaner shrimp did. Therefore, variations in client fish ectoparasite load and cleaner shrimp hunger level are two factors that affect the balance in this mutualism. Finally, our results meet some of the assumptions of biological market theory, a framework used to understand cooperative interactions, and thus this framework is suggested for future studies on this cleaning system.

Production of triploid Kuruma shrimp, Marsupenaeus (Penaeus) japonicus (Bate) nauplii through inhibition of polar body I, or polar body I and II extrusion using 6-dimethylaminopurine

This study investigated the chromosome ploidy level of Marsupenaeus (Penaeus) japonicus (Bate) non-viable (unhatched) embryos and nauplii after exposure to 6-dimethylaminopurine (6-DMAP), timed to stop either polar body (PB) I, or PBI and II extrusion. Embryos from eight separate families or spawnings were exposed to 150 or 200 mu M 6-DMAP from 1- to 3-min post-spawning detection (psd) for a 4- to 5-min duration (timed to stop PBI extrusion). Separate aliquots of embryos from five of the same spawnings were also exposed to 200 mu M of 6-DMAP from 1- to 3-min psd for a 16-min duration (timed to stop both PBI and II extrusion). For one spawning, a third aliquot of embryos was exposed to 400 p M of 6-DMAP from 1- to 3-min psd for a 16-min duration (timed to stop both PBI and II extrusion). At 18-h psd, non-viable embryo and nauplii samples were taken separately for fluorescent activated cell sorting (FACS). FACS revealed that there were diploids and triploids among all treated non-viable embryos and nauplii. All control non-viable embryos and nauplii were diploid. Percentages of triploid induction for the 4- to 5-min and 16-min durations were not significantly different (P > 0.05). Additionally, no difference was found in the triploidy level of nonviable embryos compared to nauplii in these treatments. The percentage of triploid embryos and nauplii when exposed to 6-DMAP for a 4- to 5-min duration ranged from 29.57% to 99.23% (average 55.28 +/- 5.45%) and from 5.60% to 98.85% (average 46.70 +/- 7.20%), respectively. The percentage of triploid embryos and nauplii when exposed to 6-DMAP for a 16-min duration ranged from 11.71% to 98.96% (average 52.49 +/- 11.00%) and from 47.5% to 99.24% (average 79.38 +/- 5.24%), respectively. To our knowledge, this is the first documentation of successful PBI or PBI and II inhibition in shrimp. This study conclusively shows that treatment of M. japonicus embryos with 6-DMAP at 1- to 3-min pscl for either a 4- to 5-min duration (timed to stop PBl extrusion) or 16-min duration (timed to stop both PBI and II extrusion) results in viable triploid nauplii. (c) 2006 Elsevier B.V. All rights reserved.