Controlled experiment
Direct Neocaridina evidence
Gorol et al., 2026. The European Zoological Journal 93(1): 563-582.
Source taxon as published: Neocaridina davidi
Study shape: As a continuation of the 2025 experiment, adults received clean water or nominal 1, 5 or 10 mg/L BPA for 24, 48 or 72 hours. Qualitative methods used organs from five animals per group and quantitative methods used five cell suspensions per group.
Experimental unit: Individually exposed adult and its derived organ or cell preparation; this continued the earlier exposure program and is not evidence of an independent replication
Environment: Adult shrimp held individually in 0.5-litre containers at 21 C, pH 7 and GH 10, fed ad libitum, with excrement removed every 12 hours and fresh nominal exposure water prepared for each animal
Endpoints: reactive oxygen species, apoptosis and necrosis, caspase and Bcl-2 states, DNA damage, mitochondrial structure and membrane potential, reduced glutathione.
What it can support: The assigned acute BPA treatments changed multiple intestine and hepatopancreas cell endpoints. Reactive-oxygen signals were highest after 24 hours and then declined, while cell-death, mitochondrial and DNA-damage responses varied by organ, concentration and time; hepatopancreas DNA damage was higher after 48 hours at every tested concentration.
What it cannot support: This is a continuation of the same nominal concentration and exposure design as the 2025 publication, not independent confirmation. Concentrations were not analytically verified, controls were clean-water organ groups rather than an explicitly time-matched series, and five method preparations per group do not establish the total number of independent animals across assays. Declining reactive-oxygen signal does not prove recovery because other cellular endpoints changed on different timelines. No whole-animal survival, behaviour, reproduction, chronic effect or recovery endpoint was measured. It does not establish an aquarium BPA threshold, a general plastic-safety rule or a diagnosis from visible symptoms.
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Controlled experiment
Direct Neocaridina evidence
Guz et al., 2026. Journal of Fish Diseases 49(7): e70127.
Source taxon as published: Ornamental Neocaridina spp.
Study shape: Five isolates were identified biochemically and by mass spectrometry, screened for virulence genes, challenged back into shrimp and tested for drug susceptibility.
Experimental unit: Infection challenge group or bacterial isolate, depending on endpoint
Environment: Diagnostic isolation followed by laboratory pathogenicity challenges and antimicrobial susceptibility testing
Endpoints: isolate identity, virulence genes, challenge mortality, lesion reproduction, antimicrobial susceptibility.
What it can support: The tested isolates reproduced disease in challenge animals with isolate-specific mortality and multidrug-resistance patterns.
What it cannot support: A dark lesion or lethargy cannot identify Aeromonas, and isolate resistance results do not authorize an aquarium antibiotic or dose.
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Controlled experiment
Direct Neocaridina evidence
McIvor et al., 2026. Aquaculture Research 2026(1): 6618042.
Source taxon as published: Neocaridina davidi, red cherry variant
Study shape: Three aquaria per treatment received 0, 20, 2000 or 20000 smooth 38 to 45 micrometre particles per litre, starting with 20 females and five males per aquarium.
Experimental unit: Thirty-litre aquarium, with three aquaria per concentration
Environment: Twelve 30-litre aquaria with sponge filtration and five months of smooth polystyrene-sphere exposure
Endpoints: particle ingestion, monthly mortality, counts by size class, post-exposure mortality.
What it can support: The two higher tested concentrations increased mortality and changed population size-class counts under the five-month exposure.
What it cannot support: Fecundity, gonads and individual growth trajectories were not measured, so size-class counts do not directly establish slower reproduction or individual growth rates; the treatments are not aquarium safety thresholds.
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Controlled experiment
Direct Neocaridina evidence
Gorol et al., 2025. The European Zoological Journal 92(1): 390-409.
Source taxon as published: Neocaridina davidi
Study shape: Adults received clean water or nominal 1, 5 or 10 mg/L BPA for 24, 48 or 72 hours. One animal occupied each container, and five specimens from each group were selected for each microscopy, histochemistry, fluorescence or cell-cycle method.
Experimental unit: Individually exposed adult in a 0.5-litre container; method-specific samples used five animals per group, while reuse or independence across methods was not stated clearly
Environment: Adult shrimp held individually in 0.5-litre plastic containers at 21 C, pH 7 and GH 10, fed ad libitum, with excrement removed every 12 hours and fresh nominal exposure water prepared for each animal
Endpoints: intestine and hepatopancreas ultrastructure, autophagic and acidic structures, lipid, protein and polysaccharide reserves, regenerative-cell ultrastructure, Ki67-positive cells, cell-cycle phase proportions.
What it can support: The assigned acute BPA treatments produced organ-, time- and concentration-associated changes in midgut cell structure, autophagic signals, reserve material, regenerative-cell measures and cell-cycle distributions. The highest treatment produced extensive intestinal degeneration after 72 hours.
What it cannot support: Concentrations were nominal and not analytically verified. The clean-water control came from continuous culture rather than an explicitly time-matched control series, and the number of distinct animals across the many methods was not clear. The study lasted 24 to 72 hours, used adult tissue and cell endpoints, and did not report whole-animal survival, behaviour, reproduction, recovery or a chronic exposure response. It does not establish a household BPA threshold, diagnose damage from an aquarium observation or show that an unnamed plastic product caused exposure.
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Controlled experiment
Direct Neocaridina evidence
Hugger, 2025. Master thesis, University of Vienna.
Source taxon as published: Neocaridina davidi
Study shape: Ionic silver, PVP-coated nanospheres and uncoated nanopowder were compared with measured dissolved silver and animal accumulation.
Experimental unit: Mesocosm exposure unit, not each shrimp sharing the exposure
Environment: Mesocosm exposures in artificial Danube water
Endpoints: 24-hour mortality, median lethal estimate, dissolved silver, whole-animal accumulation.
What it can support: The three tested silver forms had different dissolution and acute mortality relationships under the mesocosm chemistry.
What it cannot support: A master thesis acute result is not a chronic safe value, copper proxy, product limit or household aquarium target.
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Controlled experiment
Direct Neocaridina evidence
Ostróżka et al., 2025. The European Zoological Journal 92(1): 876-895.
Source taxon as published: Neocaridina davidi
Study shape: A continuation using the 2024 nickel experiment scheme measured cell-cycle distribution, Ki67 proliferation, ATP and ADP/ATP, mitochondrial membrane potential, HSP70 and MnSOD in intestine and hepatopancreas. Five specimens represented each control or experimental group for each qualitative and quantitative method except Western blot. The prepublication CC BY deposit supplies the mitochondrial-potential and luminometry values plus microscopy and blot images.
Experimental unit: Individually exposed adult and its isolated organ or cell preparation, with five deposited values per group and organ for mitochondrial potential, ATP and ADP/ATP. The files do not provide specimen identifiers linking endpoints or establish whether the same five animals supplied each method; whether this was a new animal cohort independent of the 2024 publication was not stated.
Environment: Adult shrimp held individually in 0.5-litre cups at 21 C, pH 7 and GH 10 during nominal 3.63 mg Ni/L exposure for one or two weeks and one or two subsequent clean-water weeks
Endpoints: cell-cycle distribution, Ki67-positive cells, ATP concentration, ADP to ATP ratio, mitochondrial membrane potential, HSP70, MnSOD.
What it can support: The one high nominal nickel treatment produced organ- and time-associated changes in proliferation, energy and mitochondrial measures. Some measures returned toward control values during clean-water periods while others did not.
What it cannot support: This publication continues the prior nickel research program and uses its experimental scheme, so it is not evidence of an independent dose replication. Water concentration was not analytically verified, the control was the continuous culture rather than a separately described time-matched cup group, recovery groups were conditional on survival and only five values represented each deposited quantitative method group. The deposit does not link values across methods or prove independent reuse of animals. It did not measure whole-animal recovery, behaviour, reproduction or a safe household concentration.
Related source material: CC BY quantitative and image dataset
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Controlled experiment
Direct Neocaridina evidence
Ostróżka et al., 2024. The European Zoological Journal.
Source taxon as published: Neocaridina davidi
Study shape: The authors first screened nominal 0.09, 0.9 and 3.63 mg Ni/L, selected 3.63 mg Ni/L after observing about 50 percent mortality at 14 days, then compared one- and two-week exposures and one- or two-week clean-water periods. Whole-body nickel, midgut structure, reserves, reactive-oxygen and cell-death measures were assessed. The linked CC BY deposit supplies quantitative tables and microscopy files.
Experimental unit: Individually held adult for exposure, with animal, whole-body preparation or isolated organ used according to endpoint. The deposited TXRF table has three numbered samples per group, while reactive-oxygen tables contain uneven row counts and repeated analysis dates that do not identify every row as an independent animal.
Environment: Adult shrimp exposed individually to nickel chloride for one or two weeks, followed by one or two weeks in clean water at 21 C, pH 7 and GH 10
Endpoints: preliminary mortality, whole-body nickel, intestinal degeneration, hepatopancreatic change, reserve material, reactive oxygen species, autophagic signals, apoptosis and necrosis, clean-water response.
What it can support: At the selected high nominal nickel treatment, adults accumulated nickel and showed organ- and time-associated cellular damage; some measured changes persisted after one or two weeks in clean water.
What it cannot support: The approximately 50 percent 14-day mortality was a preliminary selection observation, not a reported formal LC50 estimate with uncertainty. The deposit measures nickel in whole-shrimp samples, not the exposure water, so 3.63 mg Ni/L remains nominal; the recovery groups necessarily contained survivors, controls remained in the continuous culture and the repository does not resolve whether every reactive-oxygen row is a biological unit rather than a repeat or batch observation. It does not establish whole-animal recovery, a household threshold, a copper or fertilizer limit, or a diagnosis from appearance.
Related source material: CC BY quantitative and image dataset
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Controlled experiment
Direct Neocaridina evidence
Razekenari et al., 2023. Aquatic Toxicology 259: 106519.
Source taxon as published: Neocaridina davidi
Study shape: Six nominal EE2 concentrations from 0 to 200 micrograms/L were assigned to three aquaria each, with 30 females per aquarium. Ten ready females per treatment were later mated in clean water with unexposed males, eggs were removed for artificial incubation and unexposed offspring were followed for 10 days across five successive spawns.
Experimental unit: The 8-litre exposure aquarium, with three aquaria per concentration; later mating, artificial-incubation and offspring units differed by endpoint
Environment: Eighteen 15-litre laboratory aquaria with 8 litres of useful water at 26 to 28 C, complete water replacement and redosing every two days during 210 days of nominal EE2 exposure
Endpoints: female survival, female weight and length, time to first spawning, interspawn interval, fecundity, egg volume, hatching, hatchling size, 10-day offspring size and survival.
What it can support: Under this nominal exposure protocol, females in the two lowest nonzero treatments were heavier and longer but produced fewer eggs and had lower hatching outcomes, while every nonzero treatment had lower adult survival than the control.
What it cannot support: Exposure concentrations were nominal and not analytically verified. The control was clean water rather than an ethanol-matched vehicle, the written stock-volume calculation conflicts with the reported maximum ethanol percentage and control survival was only 57.3 percent. Full water was replaced every two days, males and offspring were not directly exposed and eggs were artificially incubated. The study does not establish a household threshold, a beneficial low dose, an environmental safe level or an aquarium treatment.
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Controlled experiment
Direct Neocaridina evidence
Zanitti et al., 2023. Archives of Environmental Contamination and Toxicology 85(2): 181-190.
Source taxon as published: Neocaridina davidi
Study shape: Twenty-two aquaria were randomly assigned per treatment. Each began with one ovigerous female and two mature males and received control water or nominal 0.1 or 1 mg/L diclofenac. HPLC-MS/MS checks at 0 and 72 hours in two aquaria per treatment produced overall measured means of 0.0075, 0.1320 and 1.0730 mg/L, respectively. Mortality, molts, egg loss, hatching and ovarian rematuration were checked daily; surviving adults, broods, ovaries and male distal vas deferens supplied later endpoints.
Experimental unit: The 1.5-litre aquarium, with 22 aquaria per assigned treatment
Environment: Sixty-six 1.5-litre laboratory aquaria at 27 C, pH 7.5 and hardness 80 mg/L as CaCO3, with aeration, Java moss, daily ad-libitum feed and complete water replacement plus re-dosing three times weekly during 63 +/- 3 days
Endpoints: mortality, adult specific growth rate, successive spawning timing, hatching, juvenile count and morphology, female ovarian histology, male spermatophore histology.
What it can support: Under this 63-day laboratory protocol, aggregate mortality was 22.7, 39.4 and 45.5 percent, with only the 1 mg/L nominal treatment differing significantly from control. The higher treatment reduced survivor-conditioned female growth, the proportion of females hatching their first exposed brood and the proportion of advanced ovarian oocytes, shortened interspawn timing, and increased abnormal juveniles per female in the small second-spawn sample. Incubation time, male growth, male spermatophore structure and among-treatment hatchling counts did not differ significantly.
What it cannot support: The control was not an analytical zero: its two checks were 0.009 and 0.006 mg/L, for a 0.0075 mg/L mean. The lowest nonzero measured treatment averaged 0.1320 mg/L, and only two nonzero treatments were tested. Analytical checks covered two aquaria per treatment at two times. The first egg cohort had already developed partly before exposure, later brood and tissue endpoints were conditioned on female survival and successful spawning, the second-spawn abnormality result came from few females, and male dependency may not have been fully represented by aquarium in the analysis. Stock came from a dealer-derived laboratory colony without voucher or diagnostic identity confirmation; allocation blinding, feed intake, exposure-period nitrogen chemistry, tissue residues and offspring survival or growth were not reported. The preprint abstract can overstate the juvenile-count result because the paper reports no significant among-treatment decrease. The design does not establish a trace-effect threshold, safe household medication concentration, aquarium treatment, recovery, mechanism or multigeneration outcome. The SSRN and Research Square postings are duplicate preprint versions, not independent replication.
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Controlled experiment
Direct Neocaridina evidence
Kucera et al., 2022. Fishes 7(6): 323.
Source taxon as published: Neocardina heteropoda, as spelled in the paper
Study shape: Two sequential batches each assigned three tanks to polystyrene exposure and three tanks to control water; 10 shrimp per condition were measured after 7 and 14 days in each batch.
Experimental unit: Aquarium for exposure assignment; behaviour and oxygen consumption were then measured on selected individual shrimp
Environment: Laboratory aquaria at 20 C with 0.2 to 0.5 mm virgin polystyrene particles at a nominal 8 mg/L
Endpoints: swimming speed, acceleration, distance moved, standard metabolic rate, particle presence.
What it can support: At the tested particle size and nominal concentration, exposed shrimp showed greater activity measures and lower standard metabolic rate than controls, with speed and acceleration differences appearing by day 14.
What it cannot support: The published mixed model did not list aquarium as a random effect even though exposure was assigned by tank. One high nominal concentration, short duration and virgin spherical particles do not establish a household threshold, chronic outcome or effect of other plastics.
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Controlled experiment
Direct Neocaridina evidence
Ostróżka et al., 2022. Micron 154: 103162.
Source taxon as published: Neocaridina davidi
Study shape: Midgut tissues were compared across dimethoate exposure and recovery intervals.
Experimental unit: Animal and prepared midgut tissue
Environment: One to three weeks of laboratory exposure followed by clean-water recovery
Endpoints: midgut tissue damage, cellular regeneration, recovery interval.
What it can support: The tested exposure damaged midgut tissues and recovery responses differed among organs and intervals.
What it cannot support: It does not establish that a fixed clean-water period clears an aquarium or restores every biological endpoint.
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Controlled experiment
Direct Neocaridina evidence
Klein, Hess, Nungess, Schulte-Oehlmann and Oehlmann, 2021. Environmental Science and Pollution Research 28: 62246-62254.
Source taxon as published: Neocaridina palmata, White Pearl variety as purchased
Study shape: One independent run used eight one-shrimp vessels per treatment. A 24-hour concentration series compared fluorescent polyethylene bead mixtures or milled polyvinyl chloride fragments, a separate fragment series added food, and separate groups exposed at 20000 particles/L were either sampled immediately or moved to particle-free vessels with food for four hours.
Experimental unit: Individual 500 mL vessel and shrimp; seven to eight surviving replicates per treatment in one independent run
Environment: Individual shrimp in 500 mL laboratory vessels containing reconstituted water, with particles settled before each 24-hour exposure
Endpoints: particles in rinsed whole-shrimp lysate, particles recovered from excretions, body length, sex, observed mortality.
What it can support: Under this protocol, mean particle recovery in whole-shrimp lysates rose across the 24-hour concentration series, smaller beads were recovered more often than larger beads, particles appeared in four-hour excretion samples and adding food did not produce a significant fragment-uptake difference.
What it cannot support: The source used N. palmata rather than N. davidi and did not independently verify the White Pearl trade identity. Shape was confounded with polymer, size distribution, suspension preparation and optical detection; fragments at or below 5 micrometres were excluded. The supplement reports group summaries rather than animal-level data, one incomplete nominal-versus-actual concentration example, separate highly variable immediate and four-hour groups and no outlier identities or corrected results. Three deaths across different treatments do not establish particle toxicity. Whole-animal lysate cannot localize particles to gut or tissue, and the study measured no chronic survival, growth, reproduction, histology or home-aquarium threshold.
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Controlled experiment
Direct Neocaridina evidence
Klein, Hess, Schulte-Oehlmann and Oehlmann, 2021. PeerJ 9: e12442.
Source taxon as published: Neocaridina palmata, White Pearl variety as purchased; identity was not independently verified
Study shape: Four material-by-weathering experiments each used eight one-shrimp vessels per concentration at 0.40, 1.00, 2.50, 6.25 or 15.6 g plastic equivalents/L plus medium, DMSO and lethal sodium-chloride controls. Movement was recorded on days 1, 3, 7 and 14. Separate three-run Microtox tests used the same pooled extracts.
Experimental unit: Individual 600 mL beaker containing one shrimp and 200 mL medium for the in vivo response; duplicate 24-hour leachates were combined into one extract per material and weathering condition, so shrimp vessels did not independently replicate leachate preparation
Environment: Adult shrimp held individually for 14 days in 200 mL of reconstituted water with concentrated, particle-filtered extracts from one recycled LDPE material or one starch-blend foil
Endpoints: moved distance, frozen events, daily observed mortality, daily molting, body length, sex, Aliivibrio fischeri luminescence inhibition.
What it can support: The concentrated extracts inhibited bacterial luminescence, but moved distance and frozen events in shrimp showed high individual variation, few isolated differences and no consistent concentration-response across the four material-by-weathering experiments.
What it cannot support: The in vivo treatments were particle-filtered extracts expressed as source-plastic equivalents, not particles, identified chemical doses or aquarium-product exposures. The two leachate duplicates were pooled, extracts were 5000-fold concentrated, the study did not chemically analyze its own mixtures and the lethal sodium-chloride group was excluded from the 896-trajectory analysis. Two locomotor endpoints cannot establish overall safety or absence of harm. The work did not test N. davidi, growth, reproduction, histology, chronic exposure, natural weathering or a home-aquarium threshold. Four raw XLSX supplements were available but could not be audited in this intake because the required workbook runtime was unavailable.
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Controlled experiment
Direct Neocaridina evidence
Siregar et al., 2021. Antioxidants 10(3): 391.
Source taxon as published: Research-centre animals labelled Neocaridina denticulata; no diagnostic or molecular identification method reported
Study shape: A commercial 28.8 percent imidacloprid formulation was diluted to nominal 0.03125, 0.0625, 0.125, 0.25, 0.5 and 1 ppm for up to 96 hours. The paper describes 20 shrimp in one 200 mL plastic tank per treatment and 12 animals per group for movement, heartbeat and maxilliped-movement endpoints. A separate four-group series placed 24 animals per 500 mL tank, exposed three groups to nominal 1 ppm imidacloprid for 24 hours, then transferred them to fresh water or nominal 0.1 or 1 ppm acetylcholine for three days.
Experimental unit: Exposure tank for treatment assignment, apparently one tank per concentration or rescue group; individual shrimp were later measured in multiwell plates. Repeated video recordings and animal-level measurements do not create independent treatment-tank replication.
Environment: Small plastic exposure tanks using reverse-osmosis water at pH 7.0 to 7.5 after seven days of laboratory holding at 26.5 C
Endpoints: tracked distance and immobilization, image-derived heartbeat, maxilliped movement used as a gill-ventilation proxy, whole-animal oxidative-stress and energy-metabolism kit measurements, fresh-water and acetylcholine post-exposure trajectories, surrogate-protein molecular docking.
What it can support: Under this formulation and fasting protocol, imidacloprid-treated groups showed lower tracked movement across 24 to 72 hours, and higher nominal treatments had lower heartbeat and maxilliped-movement rates at 96 hours. The small whole-animal assay series reported no group differences in its selected oxidative-stress or energy-metabolism measures.
What it cannot support: Independent exposure tanks, random allocation, formulation-only controls and measured water concentrations were not reported. The text conflicts between 12 endpoint animals and 20 exposed animals per group, and treatment is confounded with the apparent single tank while animal measurements are analysed as replicates. Starvation changed heartbeat and maxilliped movement, acetylcholine itself changed the measured endpoints, and the rescue comparison also appears to use one tank per group. Docking used a snail acetylcholine-binding protein rather than a shrimp receptor and cannot prove the in vivo mechanism. The study does not validate acetylcholine as an antidote, identify imidacloprid from behaviour, establish recovery, chronic safety, a plant or flea-product rule, an aquarium dose or a universal threshold.
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Controlled experiment
Direct Neocaridina evidence
Hu et al., 2019. Environmental Pollution.
Source taxon as published: Neocaridina davidi
Study shape: Defined fenoxycarb and methoprene exposures were compared with controls.
Experimental unit: Exposure treatment culture unit
Environment: Chronic laboratory chemical exposure
Endpoints: body length, molting frequency, cuticle-related pathways, metabolism.
What it can support: The tested insecticides changed growth, molting and molecular outcomes under the exposure design.
What it cannot support: It does not establish that every pesticide acts the same way or identify a home exposure from behaviour or molting alone.
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Controlled experiment
Direct Neocaridina evidence
Kim et al., 2018. Journal of Environmental Biology 39(1): 51-57.
Source taxon as published: Ten wild Lake Chang Pyung animals and separate exposure animals labelled Neocaridina denticulata denticulata; no diagnostic or molecular identification method reported, and the exposure-animal source was not separately stated
Study shape: A baseline whole-body cDNA library from ten non-exposed wild shrimp produced 1,247 high-quality expressed sequence tags and 603 unique sequences. A separate short-term copper experiment used three shrimp per treatment, a time series at 0, 1, 3, 6, 9 and 24 hours with the copper concentration unstated, and a 24-hour nominal dose series whose figure labels 0, 1, 2, 5, 10 and 100 micrograms per litre. Pooled whole-body RNA was assayed by real-time RT-PCR for 14 selected transcripts using beta-actin as the sole reference gene.
Experimental unit: The report does not identify the exposure container, independent vessel replication or random allocation. Three shrimp are described per treatment and their total RNA was pooled before cDNA synthesis; the triplicate experiments and figure N of 3 therefore do not clearly identify independent biological replicates.
Environment: Aerated natural lake water for the baseline cDNA library; copper-exposure conditions reported as 20 C, pH 8.0, dissolved oxygen above 5.6 mg/L and 28 PSU salinity, despite the freshwater lake animal description
Endpoints: baseline expressed sequence tags and sequence annotation, relative whole-body mRNA abundance for 14 selected targets.
What it can support: Under the reported short-term nominal copper protocol, all 14 selected transcripts varied in at least part of the time or concentration series, and nine were reported to increase by more than two-fold at one or more observations from 1 to 24 hours. This supports a candidate short-term pooled whole-body transcriptional response, not a validated field biomarker.
What it cannot support: The copper material, concentration used for the time series, exposure vessels, independent vessel replication, allocation, acclimation, feeding, water renewal and measured water concentrations were not reported. The prose loses the dose unit while the figure labels micrograms per litre, and the stated 28 PSU salinity conflicts with the freshwater lake description. Pooling makes the biological denominator unclear, beta-actin stability and PCR efficiencies were not reported, and repeated target-by-time and target-by-dose tests had no reported multiplicity model. Transcript changes do not establish protein abundance, toxicity, survival, molting impairment, immune competence, chronic harm, a safe concentration, diagnosis or validated biomarker panel.
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Controlled experiment
Direct Neocaridina evidence
Li et al., 2015. Environmental Science 36(2): 727-735.
Source taxon as published: Wild animals from vegetation around Dianshan Lake labelled Neocaridina denticulate; no diagnostic or molecular identification method reported
Study shape: Separate five-day semi-static series used nominal copper-ion treatments of 0.086, 0.172, 0.344 and 0.688 mg/L and nominal chlorpyrifos treatments of 0.0015, 0.0030, 0.0060 and 0.0120 micrograms/L. Each concentration was described as three parallel 2-litre beakers with 20 shrimp per beaker, daily solution renewal, no feeding and blank-water and chlorpyrifos vehicle controls. Three shrimp per group were destructively sampled on each day for muscle homogenate assays.
Experimental unit: The two-litre exposure beaker, described as three parallel beakers per concentration. The methods do not state how the three shrimp sampled at each time were distributed among or pooled across those beakers, although results are described as means of three parallel groups.
Environment: Two-litre beakers in temperature-controlled water baths using aerated tap water at 23 +/- 1 C, pH 7.2 to 7.7, hardness about 205.95 mg/L as CaCO3 and dissolved oxygen above 5.80 mg/L
Endpoints: five-day mortality observation, muscle protein concentration, muscle malondialdehyde content, muscle total superoxide dismutase activity, muscle acetylcholinesterase activity.
What it can support: Under the tested semi-static protocols, nominal copper-ion and chlorpyrifos concentration series were associated with time-varying muscle protein, lipid-peroxidation and enzyme-activity measurements. No deaths were reported through five days in these sublethal treatment groups.
What it cannot support: The wild-source taxon was not independently verified as N. davidi, concentrations were nominal rather than analytically measured, animals were fasted for five days and sampling-to-beaker mapping is unclear. The paper cites 96-hour LC50 values from earlier work to select its fractions but does not provide that acute experiment, uncertainty or full analysis, so those values are not results of this five-day series. Repeated t tests and one-way ANOVAs across concentration-by-day contrasts had no reported multiplicity or repeated-time model, and the text contains concentration-label inconsistencies. Muscle biomarkers do not establish a no-effect level, chronic safety, diagnosis, antidote, aquarium dose or universal copper or pesticide threshold.
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Controlled experiment
Neocaridina identity evidence
Sung et al., 2014. Environmental Toxicology and Pharmacology 38(1): 8-13.
Source taxon as published: River-collected animals from Tainan County, Taiwan, identified in the paper as Neocaridina denticulata; no voucher, morphology or molecular identification method reported
Study shape: Nominal 96-hour mortality series tested acetaminophen and ibuprofen alone at 0.2, 0.6, 1, 1.5, 2, 4 and 8 mg/L; a 1:1 series; variable acetaminophen at 0.1, 1, 2, 4 or 7.5 mg/L plus fixed 0.5 mg/L ibuprofen; and variable ibuprofen at the same levels plus fixed 0.5 mg/L acetaminophen. Stocks were prepared in DMSO. Sodium chloride at 1 to 5 g/L was used as a reference chemical, and mortality was recorded at 2, 6, 24, 48 and 96 hours.
Experimental unit: The treatment beaker, with 20 shrimp reported per group. The paper says the experiment was repeated four times and every treatment was performed in duplicate, while its mortality table reports n = 4; it does not reconcile tests, duplicate beakers and the analyzed n or explain whether beaker-level results or pooled animal counts entered each model.
Environment: Ten- to fourteen-day-old, approximately 1.5 mm juveniles from acclimated river-source stock were tested in 1-litre glass beakers under a static-renewal guide; source stock was held at 25 +/- 1 C, pH 7.4 to 7.8, dissolved oxygen above 7.3 mg/L and hardness 38 to 45 mg CaCO3/L
Endpoints: mortality time course, 96-hour LC10 with 95 percent confidence interval, 96-hour LC50 with 95 percent confidence interval, sodium-chloride reference-chemical LC50.
What it can support: Under the nominal 96-hour protocol, reported LC50 values were 6.07 mg/L for acetaminophen, 6.60 mg/L for ibuprofen, 6.23 mg/L for the 1:1 series, 4.78 mg/L for variable acetaminophen plus 0.5 mg/L ibuprofen and 6.78 mg/L for variable ibuprofen plus 0.5 mg/L acetaminophen. The experiment supports ratio-specific mortality curves under those exact series.
What it cannot support: The treatment concentrations were nominal, the paper does not report analytical verification, the DMSO fraction or a DMSO-matched control, and renewal frequency and test-time feeding are not stated. The equal-ratio LC50 does not clearly say whether concentration means each component or their sum. The unequal-series LC50 values describe the variable component while omitting the fixed 0.5 mg/L co-exposure from the reported number. The confidence intervals overlap substantially, and the paper calls patterns synergistic or antagonistic without reporting a concentration-addition, independent-action, toxicity-unit, model-deviation or isobologram analysis. The abstract says a non-significant comparison had p < 0.05. The source and identity method, allocation and duplicate-to-n mapping are incomplete. Acute mortality in tiny juveniles does not establish a safe aquarium level, chronic outcome, medication instruction, disposal rule, wastewater prediction, decontamination method or universal rule for drug mixtures.
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Controlled experiment
Direct Neocaridina evidence
Tsai and Sung, 2013. International Journal of Ecology 2(4): 38-49.
Source taxon as published: Neocaridina denticulata in the Chinese title and methods; Neocaridina denticulate in the English title
Study shape: The discovery procedure exposed shrimp to nominal 50 mg/L dipropyl phthalate for two hours followed by 12 hours in DPrP-free pond water before pooled RNA extraction. The concentration-response validation exposed groups for one day to nominal 0.5, 1 or 50 mg/L DPrP or DPrP-free pond water. DPrP stocks were prepared in acetone, but the control was not described as acetone matched. Suppression subtractive hybridization generated candidate expressed sequence tags, and 12 selected targets were checked by semi-quantitative RT-PCR.
Experimental unit: The paper describes three concentration groups and one control without reporting the number of independent exposure vessels, random allocation or a vessel-level analysis. Ten shrimp were pooled for each RNA sample, while the statistics section instead says each of at least five replicates used at least five shrimp, leaving the pooled biological-replicate construction internally inconsistent.
Environment: Aquarium-store shrimp acclimated for three days in 30 cm glass culture aquaria with pond water, fine sand, plants, an approximately 150 L/hour hang-on filter, eight hours of plant lighting and daily dried mosquito larvae
Endpoints: subtracted cDNA library, unique expressed sequence tags, semi-quantitative transcript abundance for 12 selected targets.
What it can support: The paper recovered 71 unique expressed sequence tags, including 23 matched to known-function sequences and 48 labelled unknown. Its main results, figures and summary table report that 4 of 12 selected transcripts differed from the control at nominal 0.5 mg/L, 8 at 1 mg/L and 6 at 50 mg/L after one day under the tested protocol.
What it cannot support: Concentrations were nominal and not analytically verified, acetone was not matched in the control, independent exposure-vessel replication was not reported, whole animals were pooled, control points were normalized rather than shown, and no multiplicity correction was described. The English abstract says all six responsive targets at 50 mg/L were upregulated, while the Chinese abstract, main text, figures and table show downregulation. The table footnote mistakenly names nonylphenol, and the abstract one-day discovery wording conflicts with the methods describing two hours of exposure plus 12 hours in clean water. Transcript changes do not establish protein abundance, immune competence, toxicity, survival, disease susceptibility, chronic harm, a plastic-product diagnosis, an aquarium threshold or a validated biomarker panel.
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Controlled experiment
Direct Neocaridina evidence
Liu and Sung, 2011. Bulletin of Environmental Contamination and Toxicology 87(3): 220-225.
Source taxon as published: Neocaridina denticulata
Study shape: A preliminary one-day mortality check used nominal 0.1, 0.5, 0.75 and 1 mg/L nonylphenol. Formal one-day exposures used nominal 0.001, 0.01, 0.1 and 0.5 mg/L with 60 shrimp described per 18-litre treatment aquarium and a diluted DMSO solvent control. Suppression subtractive hybridization at 0.5 mg/L generated candidate expressed sequence tags, followed by semi-quantitative RT-PCR across the concentration series.
Experimental unit: Exposure aquarium for treatment assignment and pooled whole-animal RNA sample for transcript analysis. The accessible methods describe one aquarium per concentration and do not report independently replicated treatment vessels or random allocation. At least five RT-PCR pools were made from 50 animals, so animals within a pool were not independent transcript replicates.
Environment: Twenty-litre glass aquaria with 18 litres of pond water at 28 C, pH 6.9 to 7.5, dissolved oxygen above 4 mg/L and one-day nominal nonylphenol exposure
Endpoints: preliminary one-day mortality, expressed sequence tags, semi-quantitative transcript abundance.
What it can support: After one day, 14 selected transcripts differed from the solvent control under at least one nominal nonylphenol treatment, with most reported responses being downregulation. This establishes a short-term pooled whole-animal transcriptional response under the tested laboratory protocol.
What it cannot support: Concentrations were nominal and not analytically verified. Independent exposure-vessel replication was not reported, whole animals were pooled, and no multiplicity correction was described across genes and concentrations. The abstract says six transcripts responded at 0.01 mg/L while the results text says five, and a figure labels the comparator PBS-treated even though the methods describe a DMSO solvent control. A one-day mRNA change does not establish protein abundance, functional impairment, chronic harm, recovery, behaviour, disease, reproduction, an aquarium threshold, a household product diagnosis or a biomarker protocol.
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Controlled experiment
Direct Neocaridina evidence
Huang et al., 2006. Chemosphere 64(1): 11-16.
Source taxon as published: Wild-collected Neocaridina denticulata from rivers in northern Taiwan
Study shape: Nominal chlordane treatments of 1 and 10 ng/L and lindane treatments of 0.1 and 1 microgram/L were compared with no-pesticide, acetone-vehicle and alcohol-dissolved estradiol positive controls. Female hormone-associated groups ran through 28 days. Separate groups of 20 males and 20 females per treatment were followed through oogenesis, after which each ovigerous female was moved to a 1-litre treatment beaker through hatch.
Experimental unit: The hormone-associated 10-litre exposure beaker was described as triplicate, with three shrimp pooled per sample and figure n of 15 not mapped to the beaker means. Each reproductive treatment instead used one shared 10-litre beaker, so the females and later broods were nested within a single treatment vessel.
Environment: Glass-beaker static-renewal exposure at 25 C, pH 7.4 to 7.8 and hardness 38 to 45 mg/L as CaCO3, with water renewed every 48 hours
Endpoints: estradiol assay, alkali-labile phosphate response, time to oogenesis, ovigerous rate, egg count and diameter, hatch count.
What it can support: The paper reports treatment-associated hormone-response changes and substantially reduced reproductive and hatching outcomes in several nominal chlordane and lindane groups under its exposure protocol.
What it cannot support: Exposure concentrations were nominal. Estradiol used alcohol without a described alcohol-matched control, the vehicle amount is reported in concentration units, and repeated paired t tests have no reported multiplicity correction. The reproductive comparison had one shared treatment beaker and only 0 to 8 of 20 females per group became ovigerous, making the later egg and hatch denominators small and conditional. The paper is a third distinct experiment from the same author group, not an independent replication of either 2004 cohort. It does not establish an aquarium pesticide threshold, diagnose a household event, demonstrate a mechanism or validate a treatment.
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Controlled experiment
Direct Neocaridina evidence
Huang and Chen, 2004. International Journal of Toxicology 23(2): 91-95.
Source taxon as published: Wild-collected Neocaridina denticulata from rivers in Taipei County, Taiwan
Study shape: An acute static-renewal series estimated 96-hour mortality responses to nominal chlordane and lindane concentrations. A separate 28-day experiment compared nominal chlordane at 1 and 10 ng/L and lindane at 0.1 and 1 microgram/L with no-pesticide, acetone-vehicle and alcohol-dissolved estradiol positive controls, sampling on days 1, 3, 7, 14 and 28.
Experimental unit: The exposure beaker. The acute treatments used 20 shrimp in 5-litre beakers and were described as triplicate. The chronic groups used 100 juveniles in a 10-litre beaker described as run in triplicate, but the article does not state the sampled-animal count, allocation among beakers or analysis unit.
Environment: Glass-beaker exposure after two weeks of acclimation at 25 +/- 1 C, pH 7.4 to 7.8, dissolved oxygen above 7.3 mg/L and hardness 38 to 45 mg/L as CaCO3
Endpoints: 96-hour mortality and LC50 estimate, testosterone enzyme immunoassay, alkali-labile phosphate response.
What it can support: The article reports acute mortality concentration responses, lower testosterone-associated measurements after both pesticides and a transient alkali-labile phosphate increase after chlordane under its nominal exposure protocols.
What it cannot support: The article says the acute experiment had 10 groups even though its table lists more concentration rows, prints the chlordane confidence limits in descending order and does not report measured water concentrations. Alkali-labile phosphate was an acknowledged nonspecific proxy for vitellogenin-like protein. The chronic sampling denominator and beaker-to-assay mapping are absent, estradiol used alcohol without a described alcohol-matched control, and repeated paired t tests have no reported multiplicity correction. The study does not establish a household threshold, safe concentration, mechanism, diagnosis or validated biomarker.
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Controlled experiment
Direct Neocaridina evidence
Huang, Wang and Chen, 2004. Chemosphere 57(11): 1621-1627.
Source taxon as published: Wild-collected male Neocaridina denticulata from rivers in Taipei County, Taiwan
Study shape: Nominal chlordane at 1 and 10 ng/L and lindane at 0.1 and 1 microgram/L were compared for 28 days with no-pesticide, acetone-vehicle and alcohol-dissolved estradiol positive controls. Hormone-associated samples were taken on days 1, 3, 7, 14 and 28, and masculine-appendage measurements on days 3, 7, 14 and 28.
Experimental unit: The 10-litre exposure beaker, described as triplicate for each group. The article says 150 males were used per group, while figure-level n values are 15 for hormone-associated assays and 6 for appendage measurements without explaining their distribution among the three beakers.
Environment: Ten-litre glass-beaker exposure after two weeks of acclimation at 25 +/- 1 C, pH 7.4 to 7.8, dissolved oxygen above 7.3 mg/L and hardness 38 to 45 mg/L as CaCO3
Endpoints: estradiol enzyme immunoassay, testosterone enzyme immunoassay, alkali-labile phosphate response, masculine appendage to cephalothorax length ratio.
What it can support: The paper reports treatment-associated estradiol and testosterone measurements, an alkali-labile phosphate increase only in the high chlordane group and early changes in the masculine-appendage length ratio under several nominal treatments.
What it cannot support: Water concentrations and a renewal schedule were not reported. Alkali-labile phosphate was a nonspecific proxy rather than a vitellogenin-specific assay. Estradiol used alcohol without a described alcohol-matched control. The analysis used repeated paired t tests without a reported multiplicity correction and does not explain how animal-level n values map to replicate beakers. Appendage effects varied by concentration and time and reproductive competence was not tested. The study does not validate an external-sign diagnosis, mechanism, household threshold, safe concentration or biomarker.
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