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Microparticulate feeds for marine suspension-feeders.

The development of microparticulate food particles for marine suspension-feeders is discussed with respect to the difficulties of nutrient delivery in the aquatic environment and to feeding and digestion in crustacea and bivalve molluscs. Loss of nutrients from particles suspended in seawater must be minimized by either trapping nutrients in gel particles or by encapsulation of the nutrients within impermeable walls. The gel matrix or capsule wall must be readily broken down by the animal so as to release the trapped dietary components.

Animal Feed↗

A new fixative solution to precede the reduced silver impregnation of arthropod central nervous systems.

Arthropod central nervous tissue is fixed for 1 hr at 20 C in 8% pure formic acid in 1:1 n-butanol/n-propanol prepared immediately before use (FBP), then washed for 15-30 min in 90% ethanol, and embedded in paraffin wax. Impregnation is by modified Ungewitter techniques in which the silver bath is preceded by mercury/cobalt mordanting, or by modified Holmes' methods following similar mordanting procedures. The methods yield high resolution of axons with minimal background staining, while the staining of neuronal somata is suppressed. They succeed with brains of crustacea and Odonata and other difficult materials. Tissues fixed in FBP are hard and require care in sectioning.

1-Propanol↗

Standardized white spot syndrome virus (WSSV) inoculation procedures for intramuscular or oral routes.

In the past, strategies to control white spot syndrome virus (WSSV) were mostly tested by infectivity trials in vivo using immersion or per os inoculation of undefined WSSV infectious doses, which complicated comparisons between experiments. In this study, the reproducibility of 3 defined doses (10, 30 and 90 shrimp infectious doses 50% endpoint [SID50]) of WSSV was determined in 3 experiments using intramuscular (i.m.) or oral inoculation in specific pathogen-free (SPF) Litopenaeus vannamei. Reproducibility was determined by the time of onset of disease, cumulative mortality, and median lethal time (LT50). By i.m. route, the 3 doses induced disease between 24 and 36 h post inoculation (hpi). Cumulative mortality was 100% at 84 hpi with doses of 30 and 90 SID50 and 108 hpi with a dose of 10 SID50. The LT50 of the doses 10, 30 and 90 SID50 were 52, 51 and 49 hpi and were not significantly different (p > 0.05). Shrimp orally inoculated with 10, 30 or 90 SID50 developed disease between 24 and 36 hpi. Cumulative mortality was 100% at 108 hpi with doses of 30 and 90 SID50 and 120 hpi with a dose of 10 SID50. The LT50 of 10, 30 and 90 SID50 were 65, 57 and 50 hpi; these were significantly different from each other (p < 0.05). A dose of 30 SID50 was selected as the standard for further WSSV challenges by i.m. or oral routes. These standardized inoculation procedures may be applied to other crustacea and WSSV strains in order to achieve comparable results among experiments.

Administration, Oral↗

[A method for culturing Artemia].

Artemia sp. is classified into phylum Arthropoda and class Crustacea. Ecological and experimental studies of this animal have been reported using wild species. In order to isolate inbred strains of Artemia, we have established a subculturing method for siblings of Artemia. Details of our practice on the culturing method of Artemia are described and the recent classification of Artemia is presented.

Animal Husbandry↗

Allatostatin-like immunoreactivity in the stomatogastric nervous system and the pericardial organs of the crab Cancer pagurus, the lobster Homarus americanus, and the crayfish Cherax destructor and Procambarus clarkii.

The distribution of allatostatin (AST)-like immunoreactivity was studied in the stomatogastric nervous system (STNS) and the neurosecretory pericardial organs (PO) of four decapod crustacean species by using wholemount immunocytochemical techniques and confocal microscopy. AST-like immunoreactivity was found within the STNS of all four species; its distribution in each was unique. In all four species, AST-like immunoreactivity was present in the paired commissural ganglia (CoG), in the esophageal ganglion (OG), in the stomatogastric ganglion (STG), and in their connecting nerves. Within the CoGs, numerous cell bodies and neuropil were stained. In the OG, two cell bodies were immunoreactive, although their branching pattern varies between species. In the STG of C. pagurus and H. americanus, neuropil was stained extensively, but no labeled cell bodies were found. Surprisingly, in C. destructor and P. clarkii, cell bodies were stained in the STG, one brightly stained cell body in both species and an additional two to five weakly stained cell bodies in P. clarkii. In all four species, stained gastropyloric receptor cells were present. In contrast to the variable staining within the STNS, all four species have a similar pattern of AST-like immunoreactivity within the PO. Only in C. destructor, AST-immunoreactive varicosities occur on the surface of the circumesophageal connectives and on the postesophageal commissure and suggest another neurohaemal source for AST-like peptides in this species. The pattern of this staining suggests that AST-like peptides are likely utilized as both neurohormones and as neuromodulators in the STNS of decapod crustacea.

Animals↗

Mosquito and aquatic predator communities in ground pools on lands deforested for rice field development in central Sulawesi, Indonesia.

Aquatic habitats, mosquitoes, and larvivorous predators were studied on deforested lands in Central Sulawesi, Indonesia. Open ground pools, mainly in depressions made by the treads of bulldozers and other heavy equipment, were numerous but because of their small size, comprised ca. 1% or less of the total area of the deforested lands studied. The dominant mosquitoes in these pools were Anopheles vagus, Culex vishnui, Culex tritaeniorhynchus, and Culex gelidus. The 1st 2 species were dominant in clear pools, whereas the latter 2 species were dominant in turbid pools. The dominant metazoans other than mosquitoes were Crustacea, Ephemeroptera, and Chironomidae. Both aquatic and surface predators were abundant. Dominant among aquatic predators were Anisoptera and Zygoptera nymphs, Dytiscidae, and Notonectidae. These results are discussed in relation to mosquito control on deforested lands that transitionally but inevitably appear during the course of rice field development projects in Indonesia.

Agriculture↗

Active immunisation of black tiger prawn (Penaeus monodon) against vibriosis in Thailand.

Mechanisms of host defence in Crustaceae and vibriosis in shrimp and methods for its prevention are discussed as introduction. The own work deals with the development of a site-specific multivalent anti-vibrio vaccine and its application in Thailand. The vaccine was produced in a continuous bioreactor system from field strains of Vibrio alginolyticus, V. harveyi, V. parahaemolyticus and V. vulnificus, purified through ultrafiltration, and inactivated by formalin. It was applied to Penaeus monodon PL prior to release into the ponds via artemia larvae. As mean result from numerous trials carried out under commercial field conditions in Eastern Thailand it was shown that the PL tolerated the vaccine perfectly. Due to technical problems only few of the field trials could be monitored until harvest. The data available allow claiming an obvious protective effect of the vaccine.

Animals↗

The actions of FxRFamide related neuropeptides on identified neurones from the snail, Helix aspersa.

Intracellular recordings were made from identified neurones in the suboesophageal ganglia of the snail, Helix aspersa. The actions of the eight FxRFamide analogues were investigated on these neurones. These peptides included ones isolated from arthropods and nematodes. All the peptides excited certain neurones while inhibiting others, though their relative potencies varied. Overall on neurones inhibited by these peptides the potency order was: DNFLRFamide > FMRFamide > PDVDHVFLRFamide = KNEFIRFamide > FLRFamide >> SDRNFLRFamide = SDPNFLRFamide > KHEYLRFamide. However, if the responses are compared on individual cell types, then the picture becomes more complex. For example, on cell F-2, KNEFIRFamide proved to be potent with an EC-50 value of 0.54 microM. On neurones F-13/16 and E-16, PDVDHVFLRFamide was inhibitory while FMRFamide, FLRFamide, SDRNFLRFamide and SDPNFLRFamide were excitatory. In terms of overall excitatory actions, the data are less complete but an approximate order of potency is: FMRFamide > DNFLRFamide >> SDPNFLRFamide > PDVDHVFLRFamide >> KNEFIRFamide = KHEYLRFamide = SDRNFLRFamide. However this again varies between specific neurones. These results demonstrate that peptides from insects, crustacea and nematodes are active on Helix neurones and may activate specific receptor subtypes, indicating the possible presence of endogenous analogues of these non-molluscan peptides in the Helix nervous system.

Amides↗

Methylmercury: a new look at the risks.

In the US, exposure to methylmercury, a neurotoxin, occurs primarily through consumption of fish. Data from recent studies assessing the health impact of methylmercury exposure due to consumption of fish and other sources in the aquatic food web (shellfish, crustacea, and marine mammals) suggest adverse effects at levels previously considered safe. There is substantial variation in human methylmercury exposure based on differences in the frequency and amount of fish consumed and in the fish's mercury concentration. Although virtually all fish and other seafood contain at least trace amounts of methylmercury, large predatory fish species have the highest concentrations. Concerns have been expressed about mercury exposure levels in the US, particularly among sensitive populations, and discussions are underway about the standards used by various federal agencies to protect the public. In the 1997 Mercury Study Report to Congress, the US Environmental Protection Agency summarized the current state of knowledge on methylmercury's effects on the health of humans and wildlife; sources of mercury; and how mercury is distributed in the environment. This article summarizes some of the major findings in the Report to Congress and identifies issues of concern to the public health community.

Adolescent↗

Tachykinin-related peptide and GABA-mediated presynaptic inhibition of crayfish photoreceptors.

Off-axis illumination elicits lateral inhibition at the primary visual synapse in crustacea and insects. The evidence suggests that the inhibitory action is presynaptic (i.e., on the photoreceptor terminal) and that the amacrine neurons of the lamina ganglionaris (the first synaptic layer) may be part of the inhibitory pathway. The neurotransmitters and the synaptic mechanisms are unknown. We show by immunocytochemistry that GABA and a tachykinin-related peptide (TRP) are localized in the amacrine neurons of the crayfish lamina ganglionaris. Indirect evidence suggests that GABA and TRP may be colocalized in these neurons. The extensive processes of the amacrine neurons occupy lamina layers containing the terminals of photoreceptors. Application of exogenous GABA and TRP to photoreceptor terminals produces a short-latency, dose-dependent hyperpolarization with a decay time constant on the order of a few seconds. TRP also exhibits actions that evolve over several minutes. These include a reduction of the receptor potential (and the light-elicited current) by approximately 40% and potentiation of the action of GABA by approximately 100%. The mechanisms of TRP action in crayfish are not known, but a plausible pathway is a TRP-dependent elevation of intracellular Ca(2+) that reduces photoreceptor sensitivity in arthropods. Although the mechanisms are not established, the results indicate that in crayfish photoreceptors TRP displays actions on two time scales and can exert profound modulatory control over cell function.

Animals↗

[Evaluation of genetic variability of the Kamchatka crab Paralithodes camtschaticus (Tilesius) using allozyme markers].

Interspecific genetic variation in populations of red king crab Paralithodes camtschaticus Tilesius (Litholidae, Decapoda: Crustacea) was examined using allozyme markers. The activity of 57 enzymes and the general protein presumably encoded by 92 loci was detected. The level of allozyme variability was low: the expected heterozygosity and the proportion of polymorphic loci were respectively 0.027 +/- 0.008 and 6.5%. This level of heterozygosity is three times lower than the average value for 122 crustacean species (0.082 +/- 0.007). Although genetic variants were found at 22 loci, their frequencies were generally low: only in loci 6-Pgd, Alp-1, and Pep-1 did the frequencies of the most common alleles not exceed 0.9. All polymorphic loci except one had two alleles; the exception was 6-Pgd, which had three alleles. The possible reasons for the low level of allozyme variability in red king crab are discussed.

Animals↗

[Characteristics of effects of acetylene amino alcohols M-cholinoreceptor blockers on the dopaminergic system in Daphnia magna].

Haloperidol and dimethpromide exhibit the properties of dopamine antagonists in the experiments on Daphnia magna Straus (Crustaceae family). Haloperidol, in contrast to dimethpromide, does not significantly influence the toxic effect of apomorphine. It is suggested that acetylene aminoalcohol derivatives are selective ligands for one of the subtypes of dopamine receptors.

Acetylene↗

[Probiotics--possibilities and limitations of their application in food, animal feed, and in pharmaceutical preparations for men and animals].

Probiotics are cultures of special microorganisms, which have been used as feed additives since the seventies of the past century and already since the twenties in food specimen and in pharmaceuticals. The definition of "Probioticum" was formulated in 1974 simultaneously with the use of living cultures in feed for various animals in order to substitute the application of nutritive antibiotics or chemotherapeutics. In the meantime probiotics are applied not only as feed supplements or pharmaceuticals but increasingly in suitable food specimens such as dairy products, fruit juices, chocolates, and even meat products. Of course, heating of such products prior to consumption or application must be ruled out. The selection of a suitable strain of a microorganism can be regarded as the primary requirement for the use as a probiotic. These cultures must be able to pass the stomach-duodenum barrier in a viable state and to multiply at the site of destination in the intestine. Additionally, they must be capable of producing antagonistic metabolites against a dominating saprophytic microflora resulting in a competitive growth. These abilities are common among lactic acid bacteria, e.g. lactobacilli and bifidobacteria. Yet, intended autochthonous species do not possess very good technological features for the enrichment in food specimens, because their viability will decrease rapidly under unfavourable conditions as it is the case with mineral supplements in feed or with low acidity in fermented dairy products. Therefore, some other microorganisms like spore-forming species or yeast cultures were introduced as probiotic components. These possess some similar features which render them suitable for probiotic use. But their physiological and ecological traits do not qualify them as probiotics of first choice. The special efficacy of probiotics must be strictly verified in animal nutrition due to restrictive EC-regulations, in pharmacy due to legal restraints, and in food applications in accordance with food law regulations. Safety aspects are considered very restrictively in feed applications, in the food and pharmaceutical sector they should be in accordance with the intended purpose of "fulfilling health claims". In the presented review the different requirements for the application of probiotics in animal nutrition, in food, and in pharmaceuticals will be provided. The special effect of competitive exclusion of pathogenetic and toxinogenic microorganisms in fowl performance are mentioned and even the application of probiotics as marine aquacultures of fish and Crustaceae will be included. Furthermore, the safety aspects resulting from the tremendous amount of industrially produced cultures which are distributed into the environment will be discussed. In conclusion it may be pointed out that probiotics may serve to partially replace the presently reduced or even prohibited application of nutritive antibiotics or chemotherapeutics in animal nutrition and in fulfillment of health claims in man and animals. Economic and environmental aspects will reduce the overall application of probiotics world-wide. Restrictions of use and controls of efficacy and safety are essential and must be implemented periodically.

Animal Feed↗

[Structure and function of pelagic zooplankton in various types of lakes (using the example of small lakes in northwestern Russia].

Changes of structure and functioning of pelagic zooplankton under lake eutrophication were studied in a group of small (without outlet) lakes in southern Karelia and Leningrad region. The lakes were morphologically similar and located within the same climatic zone. Differences in their trophic status were connected with anthropogenic eutrophication. Correlation between species number and feeding resources of a lake is determined as: Y = (8.01 +/- 3.85) + (0.29 +/- 0.07)X1 + (6.75 +/- 1.52)X2, r2 = 0.95, (1) Y--number of zooplankton species, X1--average chlorophyll concentration for the season, mg/1; X2--average value of biochemical oxygen demand (expressed in mgC/1). Average biomass of zooplankton for season also depends on food: logY = (-0.054 +/- 0.224) + (0.242 +/- 0.094)logX1 + (0.170 +/- 0.179)logX2, n = 13, r2 = 0.87, (2) Y--average zooplankton biomass for the season, kcal/m3, X1 and X2 the same as (1). Increase in species number and zooplankton bimass determined primarily by Rotatoria occur in parallel to chlorophyll concentration and activity of bacteria. In acid lakes rotifers are not important in energy balance. In mezotrophic lakes two main energy paths are formed--through rotifers and through Crustacea. The role of rotifers is extremely important in eutrophic lakes where about 80% of energy paths through them. The ratio community production to energy consumption for the growing season is also depended on the community structure (species number, equitability, connectence): Y = (2.257 +/- 0.026) - (0.368 +/- 0.031)X1 + (5.160 +/- 0.442)X2, r2 = 0.99, (3) Y--average seasonal production of zooplankton, kcal/m2; X1--maximal meaning of Shannon index (bites), calculated on biomass value; X2--connectence of the community, calculated according Briand (1983). Eutrophication changes the stability of lakes relative to external influences, while low productive lakes are very sensitive to the increase in nutrient load, high productive lakes are more influenced by changes in fish predation.

Animals↗

Gnathostoma infection in Nakhon Nayok and Prachin Buri, Central Thailand.

Gnathostoma infection in Nakhon Nayok and Prachin Buri Provinces, Central Thailand, was investigated. The prevalence and intensity of infection of swamp eels were determined; dog fecal samples and fresh-water copepods were examined for evidence of infection. The overall prevalence of eel infection was 38.1% (117/307) in Nakhon Nayok and 24.0% (74/308) in Prachin Buri--the former rate being significantly higher than the latter. Most of the positive Nalkhon Nayok eels (53.8%) harbored only 1-9 larvae; only one eel bore more than 50 larvae. In Prachin Buri, 67.6% of the positive eels harbored 1-9 larvae; again, only one eel bore more than 50 larvae. The mean number of 11.0 +/- 10.4 larvae/eel in Nakhon Nayok was not significantly different from that of Prachin Buri (9.3 +/- 11.4). A total of 1,292 gnathostome larvae were recovered from 307 eels in Nakhon Nayok. Of these, 52.3% had accumulated in the liver and 47.7% had spread throughout the muscles. In eels from Prachin Buri, 50.6% and 49.4% of the total of 688 larvae (from 308 eels) were found in the liver and muscles, respectively. The larvae preferred encysting in ventral of muscles rather than dorsal part; they preferred the middle portion to the anterior and posterior portions. The average length of gnathostome larvae recovered from Nakhon Nayok eels was 4.0 +/- 0.5 mm (range 2.5-5.1 mm) and the average body width was 0.40 +/- 0.05 mm (range 0.29-0.51 mm). Those from eels in Prachin Buri were 3.9 +/- 0.5 mm (range 2.2-5.1 mm) and 0.34 +/- 0.05 mm (range 0.20-0.48 mm), respectively. The mean body length and width of the larvae from eels in Nakhon Nayok were significantly greater than those of the larvae from eels in Prachin Buri. In Ban Phrao, Nakhon Nayok, none of the first 44 fecal specimens examined was positive. Of the second (68) and the third (70) specimens, one (1.5%) and two (2.9%) samples were positive. However, six months after the third fecal collection, no eggs were found. In Tha Ngam, Prachin Buri, no eggs were found in all three batches (109, 115, and 100 fecal samples). A cyclops survey of 4,000-5,000 crustacea from each of two areas (Ban Phrao and Tha Ngam) found no evidence of natural cyclops infection.

Animals↗

[The regularities in connection between fecundity with body weight and growth rate in fishes].

The absolute and relative fecundity of freshwater, anadromous and marine fishes (102 species and subspecies from 33 families) and its dependence on body mass and growth rate were analyzed on the basis of published data. According to the spawning type all studied fishes were divided into species with short-term and single spawning and fishes with extended or long-term spawning. The equations of dependence of absolute fecundity (E) on body mass (W) were calculated: E = 1.033 W0.578 (the first group) and E = 0.792 W0.74 (the second group). If W < 177 g the equations don't differ significantly and one may use the equation E = 1.34(-0.742) for both groups. The body mass of females at age of maturity expressed as a portion of maximal definitive body mass equals 0.22 +/- 0.044 for many different species with 0.95 probability. The relative fecundity (a1) of some species negatively correlates with maximal body mass of adult individuals (Wmax). This dependence is expressed by equations: a1 = 3.033 Wmax-0.549 (for the first group) and a1 = 1.726 Wmax-0.351 (for the second group). Value of ratio Wov/W of different fish species changes irregularly from 0.054 to 0.32 and its average is 0.150 +/- 0.012 for the first group and 0.156 +/- 0.007 for the second one. In such a way, single reproduction effort of fishes is approximately 0.15. Comparison of data on Pisces, Crustacea, Amphibia, Reptilia, and Mammalia revealed that reproduction effort of different aquatic and terrestrial invertebrates and vertebrates varies within rather narrow limits (from 0.05 to 0.44). Average values of this index varies even less--from 0.097 to 0.238, on average 0.162, i.e. approximately 15-18% of animals' body mass falls on their reproduction constituent.

Animals↗

Using a freshwater amphipod in situ bioassay as a sensitive tool to detect pesticide effects in the field.

In situ testing represents an alternative to conventional laboratory toxicity testing of field samples. Juvenile Paramelita nigroculus (Crustacea: Amphipoda) were exposed in situ in two rivers downstream of fruit orchard areas in the Western Cape, South Africa. Exposure took place during six time intervals (3-7 d), of which three represented the first rainfall-induced edge-of-field runoff events of the wet season, which was about two months after the last pesticide application. Survival rates were significantly reduced (35-70%) during runoff events 1 and 2 at both sites. No difference was observed from the respective no-runoff survival rate (>90%) during event 3. Peak levels of total insecticides (azinphosmethyl, chlorpyrifos, endosulfan, and prothiofos) in samples taken with water level-triggered samplers during the runoff events were between 0.03 and 0.26 microg/L in filtered water and between 305 and 870 microg/kg in suspended particles during runoff events 1 and 2 and only up to 0.01 microg/L and 101 microg/kg during event 3. Total suspended solids (TSS) varied between 400 and 700 mg/L during all three runoff events but never exceeded 65 mg/L during no-runoff time intervals. A laboratory experiment revealed that uncontaminated TSS levels of 1,500 mg/L during a 7-d exposure caused insignificant mortality (<2.5%) in P. nigroculus. No acute toxicity was observed in standard 48-h toxicity tests with juvenile Daphnia pulex using the filtered water samples taken during runoff and no-runoff conditions. It is concluded that the observed mortalities were caused by particle-associated pesticides and that the present amphipod in situ bioassay represents a sound and sensitive tool to detect runoff-related insecticide effects under field conditions.

Agriculture↗

During fictive locomotion, graded synaptic currents drive bursts of impulses in swimmeret motor neurons.

During forward swimming, motor neurons that innervate each crayfish swimmeret fire periodic coordinated bursts of impulses. These bursts occur simultaneously in neurons that are functional synergists but alternate with bursts in their antagonists. These impulses ride on periodic oscillations of membrane potential that occur simultaneously in neurons of each type. A model of the local circuit that generates this motor pattern has been proposed. In this model, each motor neuron is driven alternately by excitatory and inhibitory synaptic currents from nonspiking local interneurons. I tested this model by perturbing individual interneurons and recording synaptic currents and changes in input resistance from each class of motor neuron. I also simulated the synaptic currents that would be observed in a cell subject to different patterns of presynaptic input. When the CNS was actively expressing the swimming motor pattern, changes in the membrane potential of individual local interneurons controlled firing of whole sets of motor neurons. Membrane currents in these motor neurons oscillated in phase with the motor output from their own local circuit. The phases of these oscillations differed in different functional classes of motor neurons. In neurons that could be clamped at the reversal potential of their outward currents, the model predicted that large periodic inward currents would be recorded. I observed no signs of periodic inward currents, even when the outward currents clearly had reversed. These results permit a simplification of the cellular model. They are discussed in the context of neural control of locomotion in crustacea and insects.

Animals↗