Temperature Effects on Expression Levels of hsp Genes in Eggs and Second-Stage Juveniles of Meloidogyne hapla Chitwood, 1949.

Meloidogyne hapla is one of the most important nematode pathogens. It is a sedentary, biotrophic parasite of plants that overwinters in the soil or in diseased roots. The development of M. hapla is temperature dependent. Numerous studies have been performed on the effect of temperature on the development of M. hapla, but only a few of them analyzed the heat shock protein (hsp) genes. The aim of the study was to perform expression profiling of eight hsp genes (Mh-hsp90, Mh-hsp1, Mh-hsp4, Mh-hsp6, Mh-hsp60, Mh-dnj19, Mh-hsp43, and Mh-hsp12.2) at two development stages of M. hapla, i.e., in eggs and second-stage juveniles (J2). The eggs and J2 were incubated under cold stress (5 °C), heat stress (35 °C, 40 °C), and non-stress (10 °C, 20 °C, and 30 °C) conditions. Expression profiling was performed by qPCR. It was demonstrated that only two genes, Mh-hsp60 and Mh-dnj19, have been upregulated by heat and cold stress at both development stages. Heat stress upregulated the expression of more hsp genes than cold stress did. The level of upregulation of most hsp genes was more marked in J2 than in eggs. The obtained results suggest that the Mh-hsp90 and Mh-hsp1 genes can be used as bioindicators of environmental impacts on nematodes of the Meloidogyne genus.

The combined effect of host and food availability on optimized parasitoid life-history traits based on a three-dimensional trade-off surface.

The reproductive success of many insects is considered to be limited by two main factors: the availability of mature eggs to lay (termed egg limitation) and the time to locate suitable hosts (termed time limitation). High host density in the environment is likely to enhance oviposition opportunities, thereby selecting for higher investment in egg supply. In contrast, a shortage of food (e.g. sugar sources) is likely to increase the risk of time limitation, thereby selecting for higher allocation to initial energy reserves. To our knowledge, the combined effect of host and food availability on these optimal life-history allocations has never been investigated. We thus modelled their simultaneous effects on a three-dimensional trade-off between initial investment in energy reserves, egg number and egg size, while focusing on insect parasitoids. The model was based on Monte Carlo simulations coupled with genetic algorithms, in order to identify the optimal life-history traits of a single simulated parasitoid female in an environment in which both hosts and food are present in varying densities. Our results reproduced the simple predictions described above. However, some novel predictions were also obtained, especially when specific interactions between the different factors were examined and their effects on the three-dimensional life-history surface were considered. The work sheds light on long-lasting debates regarding the relative importance of time versus egg limitation in determining insect life-history traits and highlights the complexity of life-history evolution, where several environmental factors act simultaneously on multiple traits.

Assessment of In Ovo Administration of Bifidobacterium bifidum and Bifidobacterium longum on Performance, Ileal Histomorphometry, Blood Hematological, and Biochemical Parameters of Broilers.

Bifidobacterium is one of the most promising probiotics which was recently used as an alternative growth promoter in poultry. This trial was considered to evaluate the potential effects of in ovo administration of Bifidobacterium bifidum and Bifidobacterium longum on growth performance and biochemical parameters of broilers. Three hundred broiler breeder eggs were incubated and candled individually at day 17 of embryogenesis; then, eggs that had live embryos were randomly divided into six groups. The first and second groups were of intact (without injection) and vehicle-injected (sterile distilled water) control groups, respectively. Other groups were in ovo injected with 1 × 109 and 1 × 107 CFU B. bifidum, and 1 × 109 and 1 × 107 CFU B. longum, respectively. Hatchability was significantly (P < 0.001) affected by in ovo inoculation of bifidobacteria strains. Body weight gain and feed conversion ratio were significantly improved (P = 0.014 and 0.019, respectively) in all treatment groups. Protein fractions, aspartate aminotransferase, and alanine aminotransferase activities were not significantly altered by bifidobacteria strains in ovo injection. Regarding ileal histomorphometry parameters, a significant elevation in villus height and villus height/crypt depth ratio values were recorded while crypt depth was insignificantly affected by in ovo treatments. It could be concluded that the tested manual method of in-yolk sac injection at day 17 of incubation is more favorable in easier delivering of bifidobacteria strains which improve broiler growth performance and ileal development without any undesirable effects on blood indices and liver and renal functions.

Microbiota assessment across different developmental stages of Dermacentor silvarum (Acari: Ixodidae) revealed stage-specific signatures.

Dermacentor silvarum is an important vector of tick-borne pathogens. The microbiota of ticks has been recognized to influence their development, fitness, and reproduction as well as the acquisition, establishment and transmission of pathogens. Eggs, larvae, nymphs and adults (females and males) of D. silvarum were used in this study to evaluate microbial community and diversity across different developmental stages. The results demonstrated that some developmental stages host different bacterial species, confirming that each stage of development could have a specific associated microbiota. Proteobacteria was the dominant phylum in most stages, while Actinobacteria was the most abundant in nymphs. The abundance of Brevibacterium significantly increased from the egg stage to the following stages, and there was a sharp increase in the abundance of this genus among nymphs. Eggs showed the highest microbial richness and diversity, and the microbial community of eggs exhibited high similarity to that of females. Moreover, the high abundance of Coxiella in eggs and females further indicated that members of this genus are vertically transmitted symbionts. Nymphs showed a microbial composition distinct from those of the other stages, and the microbial community associated with males possessed higher species richness and greater bacterial diversity than that of females. Further investigations are needed to determine the functions of the microbiota and provide information enabling a better understanding of the diversity of the microbial community at different life stages.

A pulsed-dose study evaluating chronic toxicity of chlorothalonil to fish: A case study for environmental risk assessment.

Chlorothalonil is a fungicide which is highly toxic to aquatic organisms. However, in natural aquatic environments, it is very rapidly degraded, with a half-life typically in hours, reducing exposure of aquatic organisms and the potential for effects. In standard regulatory studies looking at the chronic toxicity of chlorothalonil to fathead minnow, the most sensitive endpoint was fecundity. A standard fish full-life cycle study, where chlorothalonil concentrations were maintained constant throughout, resulted in a no-observed-effect concentration (NOEC) of 1.4 µg/L. Comparing peak modeled exposure concentrations to this NOEC can result in the chronic risk to fish being considered unacceptable. The present study investigated the effect on fecundity in fathead minnow using a fish short-term reproduction assay. Five different exposure profiles were employed with time-varying concentrations based on realistic worst-case modeled environmental exposure profiles, multiplied by an assessment factor of 10, which resulted in maximum measured concentrations up to 15.5 µg/L. There were no effects on fecundity from any of the exposure profiles tested. Therefore, based on these more realistic exposure profiles, the chronic risk to fish could be considered acceptable if these exposures were deemed to be representative of the worst case. Environ Toxicol Chem 2019;38:1549-1559. © 2019 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals, Inc. on behalf of SETAC.

Allometry of individual reproduction and defense in eusocial colonies: A comparative approach to trade-offs in social sponge-dwelling Synalpheus shrimps.

Eusociality, one of the most complex forms of social organization, is thought to have evolved in several animal clades in response to competition for resources and reproductive opportunities. Several species of snapping shrimp in the genus Synalpheus, the only marine organisms known to exhibit eusociality, form colonies characterized by high reproductive skew, and aggressive territoriality coupled with cooperative defense. In eusocial Synalpheus colonies, individual reproduction is limited to female 'queens', whose fecundity dictates colony growth. Given that individual reproduction and defense are both energetically costly, individual and colony fitness likely depend on the optimal allocation of resources by these reproducing individuals towards these potentially competing demands. Synalpheus species, however, display varying degrees of eusociality, suggesting that reproducing females have adopted different strategies for allocation among reproduction and defense. Here, we use structural equation modeling to characterize the relationships between the allometry of queen reproductive capacity and defensive weaponry, and colony size in six eusocial Synalpheus species, estimating trade-offs between reproduction and defense. We document strong trade-offs between mass of the fighting claw (defense) and egg number (reproduction) in queens from weakly eusocial species, while the trade-off is reduced or absent in those from strongly eusocial species. These results suggest that in less cooperative species, intra-colony conflict selects for queen retention of weapons that have significant costs to fecundity, while reproducing females from highly eusocial species, i.e., those with a single queen, have been able to reduce the cost of weapons as a result of protection by other colony members.

Attempt to Develop Cost-Effective Rearing of Amblyseius swirskii (Acari: Phytoseiidae): Assessment of Different Artificial Diets.

Commercial production of Amblyseius swirskii Athias-Henriot based on storage mites needs both space and labor to maintain large cultures of these prey, and also may lead to health problems for workers. Therefore, the accessibility of a suitable artificial diet could eliminate the mentioned problems; however, the artificial diets must support the persistent production of high quality progeny. This study endeavored to find a more easily available and cheaper nutrient that may further reduce the cost of diet production for A. swirskii. The predator's performance was determined when it was fed on a basic artificial diet (AD1) composed of honey, sucrose, tryptone, yeast extract, and hen egg yolk, and on eight other artificial diets consisting of 80% AD1 enriched with different nutrients including maize pollen (AD2), hemolymph of Plusia gamma L. (AD3), Ephestia eggs (AD4), Artemis cysts (AD5), Ephestia last-instar larvae (AD6), multivitamin syrup (AD7), bovine serum albumin (AD8), and bull sperm (AD9). The lowest development time was on AD2. The highest value of fecundity and oviposition period were observed on AD5, followed by AD2 and AD4. The intrinsic rate of increase (r) and the finite rate of increase (λ) reached the maximal value on AD5. Feeding on AD2 and AD5 resulted in highest value of R0 (net reproductive rate). Our results indicated that Artemia cysts and maize pollen had better potential to be used as nutrient in artificial diet for mass production of A. swirskii. Overall, it seems that AD2 is the most cost effective than others.

Viability assessment of Ascaris suum eggs stained with fluorescent dyes using digital colorimetric analysis.

The aim of the study was to develop a method for the colorimetric evaluation of nematode eggs using appropriate instruments. The materials for the study were live and dead (inactivated) eggs of the Ascaris suum. Viability of the eggs was assessed using four different kits for fluorescent staining (for each technique, a series of photos were taken). Images of stained eggs were analysed using graphic software with RGB (red-green-blue) function. The viability of the eggs was assessed according to the relative positions of the distributions of colour intensities of live or dead eggs - distributions area's overlap index (DAOI), and distributions area's separation index (DASI) were calculated. Computer analysis of the intensity of green colour was not satisfactory. However, analysis of images in the spectrum of red colour proved useful for the effective differentiation between live or dead eggs. The best parameters were observed using the Annexin V FITC Apoptosis Detection Kit (DASI = 41 and 67). The investigation confirmed the usefulness of fluorescent dyes used in conjunction with digital analysis for the assessment of the viability of A. suum eggs. The use of computer software allowed a better objectivity of the assessment, especially in the case of doubtful staining.

Spirotetramat Resistance Selected in the Phenacoccus solenopsis (Homoptera: Pseudococcidae): Cross-Resistance Patterns, Stability, and Fitness Costs Analysis.

The Phenacoccus solenopsis Tinsley (Homoptera: Pseudococcidae) is a major agricultural and horticultural pest of crops throughout the world. To develop a better resistance management strategy for P. solenopsis, we conducted a study on life history parameters of different populations of this pest, one selected with spirotetramat (Spiro-SEL), an unselected (UNSEL) population, and their reciprocal crosses. We also studied the cross-resistance and the stability of spirotetramat resistance. The Spiro-SEL of P. solenopsis exhibited a 328.69-fold resistance compared to the susceptible population (Lab-PK). The Spiro-SEL population also displayed a moderate level of cross-resistance to profenofos and bifenthrin and a high level of cross-resistance to abamectin. Resistance to spirotetramat in Spiro-SEL was unstable in the absence of selection. The study of life history parameters showed that there was a significant reduction in fitness parameters of Spiro-SEL population with a relative fitness value of 0.14. There was a significant decrease in survival rate, pupal weight, fecundity, egg hatching percentage, male and female generation time, intrinsic rate of population increase of males and females, biotic potential, and mean relative growth rate. It is concluded that selection with spirotetramat had marked effect on resistance development in P. solenopsis and upon removal of selection pressure spirotetramat resistance declined significantly, indicating unstable resistance. Development of resistance led to high fitness costs for the spirotetramat-selected population. Our study may provide the basic information on spirotetramat resistance and its mechanism to help develop the resistance management strategies.

Assessment of in vitro and in vivo anthelminthic potential of extracts of Allium sativum bulb against naturally occurring ovine gastrointestinal nematodiosis.

BACKGROUND: The rapid development of anthelminthic resistance has limited the success of traditional control programmes, thereby forcing researchers to search for ethno-veterinary alternatives. OBJECTIVE: The objective is to assess the anthelminthic potential of various extracts of the bulb of Allium sativum in naturally infected sheep. ANIMALS AND METHODS: In vitro anthelminthic activities of crude aqueous and methanolic extracts of the bulb of A. sativum were investigated against the egg (500 eggs/ml) and larvae of naturally infected sheep. The aqueous extract of A. sativum was also investigated for in vivo anthelminthic activity in three groups (n = 15 each) of naturally infected Chokla sheep with a negative control group receiving no treatment, a positive control group was given a single oral dose of albendazole at 7.5 mg/kg bodyweight, and a group administered a single oral dose of an aqueous extract at 5 g/animal. Data were analysed using the general linear model. RESULTS: Aqueous extract showed better efficacy in egg hatch assay and larval development test. However, in larval paralysis test, reverse trend was seen as methanolic extract was more potent than the aqueous counterpart. A significant amount of 57% faecal egg count reduction was observed in in vivo trail using the aqueous extract on day 21 post-treatment, although in initial stages it showed 30% and 83% effectiveness on days 7 and 14 post-treatment, respectively. No deleterious ill effect was found in any of the haematological and biochemical parameters. CONCLUSIONS: Bulb of A. sativum possesses good anthelminthic efficacy and further research is thereby warranted before recommending it for nematode control programme in ovines.

Effects of spirotetramat on Aonidiella aurantii (Homoptera: Diaspididae) and its parasitoid, Aphytis melinus (Hymenoptera: Aphelinidae).

Laboratory and field studies were conducted to measure the effects of spirotetramat on life stages of California red scale, Aonidiella aurantii (Maskell), and a primary parasitoid, Aphytis melinus DeBach. Organophosphate-resistant and -susceptible populations responded similarly to spirotetramat, suggesting there is no cross-resistance between these insecticide classes. First and second instar male and female A. aurantii were 10- and 32-fold more susceptible to spirotetramat (LC50 = 0.1-0.2 ppm) compared with early third (LC50 = 1.5 ppm) and late third instar females (LC50 = 5.3 ppm). The LC99 value indicated that late stage third instar females would not be fully controlled by a field rate of spirotetramat; however, spirotetramat would reduce their fecundity by 89%. Field applications of spirotetramat in two water volumes and using two adjuvants (oil and a nonionic spray adjuvant) showed similar reduction in A. aurantii numbers, even though the higher water volume demonstrated more complete coverage. These data suggest that this foliarly applied systemic insecticide can be applied in as little as 2,340 liters/ha of water volume, minimizing application costs, and that the two adjuvants acted similarly. The endoparasitoid, A. melinus, was unaffected by the field rate of spirotetramat when it was applied to the host when the parasitoid was in the egg or larval stage. Adult A. melinus showed 2 wk of moderate reductions in survival when exposed to leaves with field-weathered residues. Spirotetramat is an integrated pest management compatible insecticide, effective in reducing A. aurantii stages and allowing survival of its primary parasitoid A. melinus.

Western bean cutworm survival and the development of economic injury levels and economic thresholds in field corn.

Western bean cutworm, Striacosta albicosta (Smith) (Lepidoptera: Noctuidae), is a native pest of dry beans (Phaseolus vulgaris L.) and corn (Zea mays L.). Historically, the western bean cutworm was distributed in the western United States, but since 1999 eastward expansion has been observed. In corn, economic impact is caused by larval ear feeding. Information on western bean cutworm biology, ecology, and economic impact is relatively limited, and the development of economic injury levels (EILs) and economic thresholds (ETs) is required for more effective management. Studies during 2008-2011, across three ecoregions of Nebraska, sought to characterize western bean cutworm survival and development of EILs and ETs. Calculations of EILs and ETs incorporated the dynamics of corn price, management cost, and pest survival. The results from the current study demonstrated low larval survival of this species (1.51-12.82%). The mean yield loss from one western bean cutworm larva per plant was 945.52 kg/ha (15.08 bu/acre), based on 74,100 plants per ha. Economic thresholds are expressed as a percentage of plants with at least one egg mass. This study is the first study that explicitly incorporates variable management costs and crop values into western bean cutworm EIL calculations, and larval survival into ET calculations.

Susceptibility of Tribolium castaneum life stages exposed to elevated temperatures during heat treatments of a pilot flour mill: influence of sanitation, temperatures attained among mills floors, and costs.

The influence of sanitation on responses of life stages of the red flour beetle, Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae), was investigated in a pilot flour mill subjected to three, 24-h heat treatments by using forced-air gas heaters fueled by propane. Two sanitation levels, dusting of wheat flour and 2-cm-deep flour, were created in 25 plastic bioassay boxes, each holding eggs, young larvae, old larvae, pupae, and adults of T. castaneum plus two temperature sensors. Data loggers (48) were placed on the five mill floors to record air temperatures. The time required to reach 50 degrees C, time above 50 degrees C, and the maximum temperature among mill floors and in bioassay boxes were measured. The maximum temperature in bioassay boxes and in the mill was lower on the first floor than on other floors. This trend was apparent in time required to reach 50 degrees C and time above 50 degrees C, especially in compartments with 2-cm-deep flour. The mean +/- SE mortality of T. castaneum life stages on the first floor was 55.5 +/- 12.9-98.6 +/- 0.8%; it was 93.2 +/- 6.7-100 +/- 0.0% on other floors. Adults were the least susceptible stage. Mortality of T. castaneum stages in compartments with 2-cm-deep flour was generally lower than those with flour dust. Costs for the three heat treatments ranged from US$27,438 to $28,838. An effective heat treatment can be conducted within 24 h, provided temperatures on mill floors reach 50 degrees C in 8-12 h and are held above 50 degrees C for at least 10-14 h, with maximum temperatures held between 50 and 60 degrees C.

Brood parasite eggs enhance egg survivorship in a multiply parasitized host.

Despite the costs to avian parents of rearing brood parasitic offspring, many species do not reject foreign eggs from their nests. We show that where multiple parasitism occurs, rejection itself can be costly, by increasing the risk of host egg loss during subsequent parasite attacks. Chalk-browed mockingbirds (Mimus saturninus) are heavily parasitized by shiny cowbirds (Molothrus bonariensis), which also puncture eggs in host nests. Mockingbirds struggle to prevent cowbirds puncturing and laying, but seldom remove cowbird eggs once laid. We filmed cowbird visits to nests with manipulated clutch compositions and found that mockingbird eggs were more likely to escape puncture the more cowbird eggs accompanied them in the clutch. A Monte Carlo simulation of this 'dilution effect', comparing virtual hosts that systematically either reject or accept parasite eggs, shows that acceptors enjoy higher egg survivorship than rejecters in host populations where multiple parasitism occurs. For mockingbirds or other hosts in which host nestlings fare well in parasitized broods, this benefit might be sufficient to offset the fitness cost of rearing parasite chicks, making egg acceptance evolutionarily stable. Thus, counterintuitively, high intensities of parasitism might decrease or even reverse selection pressure for host defence via egg rejection.

Broodstock diet effect on sea urchin Paracentrotus lividus (Lamarck, 1816) endotrophic larvae development: potential for their year-round use in environmental toxicology assessment.

The effect of captive broodstock diet on fertilization and endotrophic larvae development of the sea urchin Paracentrotus lividus was assessed. Maize grain and five inert pelleted diets were tested, during a three-month experimental period. Maize flour, wheat flour, soybean flour, maize/wheat flour (MWF) and maize/soybean flour mixes were used as vegetal sources for inert feed. Gonad index, percent egg fertilization and larvae malformation occurrence were compared with the results obtained from wild sea urchins (W). Whole egg total amino acid composition was concomitantly analyzed as a tool to explain eventual endotrophic larvae malformations caused by lack of specific nutrients. For all treatment groups (wild and captive), percent egg fertilization values above 96% were always observed, fulfilling the requisites (70-90%) necessary to conduct environmental monitoring bioassays, according to USEPA (2002). Similar values for normal percent larval development were only obtained from P. lividus broodstock subjected to an inert feeding diet based on a maize/wheat flour mix (85.0±1.45%), in comparison to wild P. lividus (82.5±1.75%). Likewise, no statistical differences on resultant whole egg total amino acid composition were observed between P. lividus fed MWF and wild treatments. Moreover, statistical differences between MWF and all the other captive feeding treatments were found for six out of the seventeen amino acids analyzed. This study demonstrates the possibility to obtain high values for P. lividus endotrophic larvae percent normal development based on broodstock held in captivity as long as an appropriate inert diet is provided.

Cost-effective binomial sequential sampling of western bean cutworm, Striacosta albicosta (Lepidoptera: Noctuidae), egg masses in corn.

Striacosta albicosta (Smith) (Lepidoptera: Noctuidae), is a native pest of dry beans (Phaseolus vulgaris L.) and corn (Zea mays L.). As a result of larval feeding damage on corn ears, S. albicosta has a narrow treatment window; thus, early detection of the pest in the field is essential, and egg mass sampling has become a popular monitoring tool. Three action thresholds for field and sweet corn currently are used by crop consultants, including 4% of plants infested with egg masses on sweet corn in the silking-tasseling stage, 8% of plants infested with egg masses on field corn with approximately 95% tasseled, and 20% of plants infested with egg masses on field corn during mid-milk-stage corn. The current monitoring recommendation is to sample 20 plants at each of five locations per field (100 plants total). In an effort to develop a more cost-effective sampling plan for S. albicosta egg masses, several alternative binomial sampling plans were developed using Wald's sequential probability ratio test, and validated using Resampling for Validation of Sampling Plans (RVSP) software. The benefit-cost ratio also was calculated and used to determine the final selection of sampling plans. Based on final sampling plans selected for each action threshold, the average sample number required to reach a treat or no-treat decision ranged from 38 to 41 plants per field. This represents a significant savings in sampling cost over the current recommendation of 100 plants.

Hatching egg characteristics, chick quality, and broiler performance at 2 breeder flock ages and from 3 egg weights.

The objective of this study was to determine the effects of flock age and egg weight on hatching egg characteristics, fertility, hatchability, salable chick production, and broiler performance using a commercial Cobb 500 broiler breeder flock. Hatching eggs from the same breeder flock in 3 weight categories (light, average, and heavy) were obtained from a commercial hatchery when the birds were 29 and 59 wk of age. One group of eggs per age and weight category was selected to assess specific gravity and was broken open to weigh egg components. Another group of eggs was incubated for 21.5 d and incubation parameters were measured. At hatching, all salable chicks were individually weighed and placed in floor pens, where they were grown out for 41 d. Daily mortality, weekly feed consumption, and individual BW at 21 and 41 d were recorded. Irrespective of flock age and egg weight, all eggs had a specific gravity lower than 1.080, the commercial set value. Eggs from the young flock age had a smaller proportion of yolk and a greater proportion of albumen. Age affected fertility, with a lower value observed at 29 wk of age. Chicks from the flock at 59 wk hatched earlier than chicks from the flock at 29 wk, and light eggs hatched earlier than both average and heavy eggs. Broilers from the 29-wk-old breeders had a lower final BW than broilers from the 59-wk-old breeders. The smaller proportion of yolk in eggs from 29-wk-old broiler breeders may be associated with the low final BW observed in their offspring. This could mean that chicks hatching from young broiler breeders, which produce eggs with small yolks, may be at a disadvantage when reared under the same conditions as chicks produced by older broiler breeders whose eggs have larger yolks.

Methodological basis for the optimization of a marine sea-urchin embryo test (SET) for the ecological assessment of coastal water quality.

The sea-urchin embryo test (SET) has been frequently used as a rapid, sensitive, and cost-effective biological tool for marine monitoring worldwide, but the selection of a sensitive, objective, and automatically readable endpoint, a stricter quality control to guarantee optimum handling and biological material, and the identification of confounding factors that interfere with the response have hampered its widespread routine use. Size increase in a minimum of n=30 individuals per replicate, either normal larvae or earlier developmental stages, was preferred to observer-dependent, discontinuous responses as test endpoint. Control size increase after 48 h incubation at 20 degrees C must meet an acceptability criterion of 218 microm. In order to avoid false positives minimums of 32 per thousand salinity, 7 pH and 2mg/L oxygen, and a maximum of 40 microg/L NH(3) (NOEC) are required in the incubation media. For in situ testing size increase rates must be corrected on a degree-day basis using 12 degrees C as the developmental threshold.

[Standardization of larval development of the sea urchin, Paracentrotus lividus, as tool for the assessment sea water quality]. / Standardisation du développement larvaire de l'oursin, Paracentrotus lividus, pour l'évaluation de la qualité d'une eau de mer.

All stages of development of the sea urchin are of interest in ecotoxicology; the largest number of prior works concerns studies on gametes. Previous studies indicated that the use of sea urchin larvae was difficult because of the need to obtain the parent generations and good quality gametes. Progress in sea urchin culture has allowed one to standardize the method. The proposed technique is based on an evaluation of the number of non-developing fertilized eggs, on the frequency of malformations, and on the length of the larvae at 96 hours, using parents raised under well-controlled conditions. Temperature (18-22 degrees C), salinity (28-34 ppt) and pH (8-8.4) have been fixed to standardize the proposed biological test. Thirty micrograms per litre of copper reduce significantly the length of the larvae and could be used as a positive control. On the other hand, reconstituted sea water permits an optimal development of the larvae and may be used as negative control. A seasonal follow-up of water quality has been achieved to validate the use of this technique in a surveillance network of water quality. The method may be used whatever the salinity, including fresh and brackish waters.