Controlled experiment
Direct Neocaridina evidence
Antcliffe et al., 2026. Royal Society Open Science 13(3): 251712.
Source taxon as published: Neocaridina davidi, with Palaemon varians as a marine comparison
Study shape: Oxygen saturation was followed for mostly 48 hours in open and closed carcass containers with carcass-free controls. A separate group of freshwater carcasses was scored daily for ten days for internal and external post-mortem change.
Experimental unit: Individual adult carcass in one container: three open and five closed freshwater oxygen containers; controls were carcass-free containers but their per-condition allocation was not clearly reported. A separate 12-carcass freshwater sequence supplied daily morphology scores.
Environment: Individual euthanized adult carcasses in 20 mL closed or 100 mL open reverse-osmosis deionized freshwater at 21 C, plus a separate open-water anatomical decay sequence
Endpoints: oxygen saturation near or around the carcass, hourly oxygen change, post-mortem internal and external morphology scores.
What it can support: In the tested 20 mL closed freshwater vials, a single adult N. davidi carcass drove oxygen saturation to complete drawdown within about one day. Open 100 mL freshwater bottles remained above full anoxia but every carcass run became at least dysoxic by 48 hours. Separate daily scoring showed rapid internal tissue opacity before major external collapse.
What it cannot support: The one-carcass-per-20-or-100-mL reverse-osmosis-water systems had no filter, substrate, plants, aeration, scavengers or living tankmates and cannot be scaled to a home aquarium, a tank-wide oxygen prediction or a removal deadline. Oxygen was reported as percent saturation and measured locally or in the tiny vial, not as aquarium mg/L. Freshwater oxygen groups were small, with three open and five closed containers, control allocation was unclear, oxygen and anatomy sequences used different euthanasia methods, and the work did not measure disease transmission, tankmate outcomes or cause of death. Clove oil and magnesium chloride methods are not home euthanasia protocols.
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Controlled experiment
Neocaridina identity evidence
Feng et al., 2026. PLOS ONE 21(2): e0342746.
Source taxon as published: Neocaridina denticulata sinensis
Study shape: The NdTryp gene was cloned, expression was profiled across tissues and nine embryonic stages, in situ hybridization localized transcripts, RNA interference was followed by injected Vibrio parahaemolyticus challenge, and recombinant protein activity was tested in vitro.
Experimental unit: Individual shrimp for animal endpoints, usually three animals per group or time point; the wording for pooled embryonic samples does not clearly separate the three individuals from the stated three biological replicates, and recombinant-enzyme measurements used technical triplicates
Environment: Market-sourced shrimp acclimated for seven days in recirculating laboratory aquaria at 25 C, followed by molecular assays, injected bacterial challenge and isolated recombinant-protein tests
Endpoints: tissue and developmental expression, hepatopancreatic localization, RNA interference knockdown, histopathology after bacterial injection, recombinant enzyme activity.
What it can support: NdTryp expression was highest in the hepatopancreas and localized mainly to R cells and epithelial cells lining its tubules. Expression appeared late in embryonic development, rose after the tested bacterial injection, and knockdown before challenge was associated with more severe hepatopancreatic damage than the dsEGFP challenged control.
What it cannot support: The injected laboratory challenge, small animal groups and ambiguous embryonic pooling do not establish natural infection, diagnosis, treatment, survival or pathogen clearance. The recombinant enzyme optima and 20 mM ion assays are not aquarium pH, temperature, mineral, copper or cadmium targets. No feeding trial tested trypsin as an additive, digestibility aid or growth treatment, and no explicitly described uninfected RNA interference histology control isolates knockdown injury from infection interaction.
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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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Review or synthesis
Broader ornamental-shrimp review
Holroyd et al., 2026. Conservation Physiology 14(1): coag046.
Source taxon as published: Tropical ornamental shrimp, including Neocaridina literature
Study shape: Published physiology, trade and transport evidence was synthesized and major gaps were identified.
Experimental unit: Published source, not a new animal experiment
Environment: Review of aquarium trade and transport evidence
Endpoints: temperature evidence, oxygen evidence, carbonate chemistry, nitrogen waste, health assessment.
What it can support: The review maps major species-specific evidence gaps and relevant transport stressors.
What it cannot support: It does not validate a drip rate, package, forecast gate, water target or treatment for Neocaridina.
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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
Chen et al., 2025. Antioxidants 14(6): 635.
Source taxon as published: Neocaridina denticulata
Study shape: One hundred twenty juveniles were assigned to a basal diet or feed containing 0.4, 4 or 40 micrograms methyl farnesoate per kilogram, with repeated length measurements and end-point biochemical and transcriptomic assays.
Experimental unit: Not recoverable as an independent tank unit: the paper describes four diet groups but does not report replicate tanks; RNA sequencing used three individual shrimp per group
Environment: Forty-day laboratory feeding study at 26 C with a 14-hour light and 10-hour dark cycle and recirculating filtration
Endpoints: body length growth rate, digestive enzyme activity, oxidative-stress assays, whole-animal gene expression.
What it can support: The reported length, enzyme, oxidative-stress and gene-expression outcomes varied with dietary methyl farnesoate concentration and time in the study population.
What it cannot support: The paper does not describe replicated tanks, so diet may be confounded with one group or culture system and individual shrimp do not repair that treatment-level replication problem. The control feed is not explicitly described as ethanol-vehicle matched, and prepared-feed hormone concentration, stability, leaching, intake and internal dose were not verified. Survival, reproduction, long-term safety and withdrawal were not established. This study cannot support a home additive, water dose, growth product or breeding treatment.
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Controlled experiment
Neocaridina identity evidence
Feng et al., 2025. Comparative Immunology Reports 8: 200226.
Source taxon as published: Market-source Neocaridina denticulata sinensis without reported voucher or diagnostic identity confirmation
Study shape: The NdCrus1 cDNA was cloned, transcript abundance was compared across ten tissues from three healthy shrimp, recombinant NdCrus1 was expressed in Escherichia coli, and the purified protein was tested against bacterial growth, selected proteases, chitin materials, reducing conditions and heat pretreatment.
Experimental unit: Individual shrimp or tissue RNA for expression work and a bacterial culture, protein reaction or material-binding preparation for in vitro endpoints; three replicates were commonly reported, but biological and technical replication was not mapped clearly for every assay
Environment: Market-source shrimp held at 25 C in aerated circulating water, followed by tissue expression work and isolated recombinant-protein assays
Endpoints: tissue transcript abundance, bacterial growth and inhibition zones, purified and bacterial protease inhibition, chitin and chitosan binding, activity after reducing-agent exposure, activity after purified-protein heat pretreatment.
What it can support: NdCrus1 was cloned and its recombinant protein inhibited proteinase K and proteases in selected bacterial preparations, showed qualitative binding to chitin materials, and lost inhibitory activity after stronger reducing or heat pretreatments. It did not significantly inhibit growth of Bacillus subtilis or Vibrio parahaemolyticus in the reported bacterial-growth assay.
What it cannot support: The study did not challenge living shrimp, manipulate NdCrus1 in living shrimp or measure survival, clinical signs, pathogen load, clearance, transmission or protection. An antimicrobial-peptide family label and in vitro protease inhibition do not establish disease resistance, diagnosis or treatment. Qualitative chitin and chitosan binding does not validate a feed, prebiotic, supplement, molting aid or dose. The 30 to 100 C pretreatments were applied to purified recombinant protein and are not shrimp temperature-tolerance or aquarium-care tests. Tissue expression used only three animals and the ovary and testis sampling allocation was unclear. No voucher, morphological diagnosis or molecular identity confirmation was reported for market-source animals.
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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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Morphology or taxonomy
Direct Neocaridina evidence
Kusmintarsih et al., 2025. E3S Web of Conferences 609: 02003.
Source taxon as published: Market-labeled Neocaridina denticulata and Neocaridina palmata; no host-identification method reported
Study shape: One hundred market-labeled N. denticulata and 100 market-labeled N. palmata were transported in market aquarium water. Gill and cephalothorax material was scraped onto slides and examined by microscopy for organisms reported as sessile bdelloid rotifers, Vorticella sp. and Zoothamnium sp.
Experimental unit: Individual examined shrimp, clustered within an unreported number of sampled aquariums from one seller and one claimed river source
Environment: Two hundred shrimp sampled across more than eight aquariums operated by the only ornamental-shrimp seller at one fish market in Purbalingga, Central Java, Indonesia
Endpoints: presence of reported organism group, host label, reported sex, gill or cephalothorax location.
What it can support: In this one-seller sample, the authors reported at least one of the three organism groups on 156 of 200 examined shrimp, including 78 of 100 under each market species label.
What it cannot support: Aquarium identities and sample counts, collection dates, host-species identification, sexing method, microscope magnification, diagnostic key, stain, image vouchers, molecular confirmation, examiner blinding, confidence intervals and the named statistical test were not reported. The two market labels were reportedly kept together, and the analysis did not account for aquarium clustering. No tissue damage, respiration, behaviour, growth, reproduction, mortality, transmission, treatment or causal disease endpoint was measured. The 78 percent result is not a trade-wide prevalence estimate, does not validate a home diagnosis, does not prove every detected organism was parasitic or harmful and does not establish a treatment indication.
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Controlled experiment
Direct Neocaridina evidence
Kusuma et al., 2025. Juvenil 6(3): 290-300.
Source taxon as published: Neocaridina sp., laboratory reproduction-stock animals without species, line or identification method reported
Study shape: Tubifex sp., Chironomus larvae, Daphnia sp. or an unidentified Spirulina pellet control were assigned to three aquaria each. The paper states one shrimp per litre, one week of acclimation and ad-libitum availability checked twice daily, with old food replaced when present.
Experimental unit: Aquarium assigned to feed, with three aquaria per treatment; the exact integer starting count is not stated, because the reported 21.6-litre volume and one-animal-per-litre density do not resolve it, and endpoint animal or aquarium denominators are not supplied
Environment: Twelve nominal 21.6-litre aquaria at approximately 25.49 to 26.86 C, pH 6.34 to 7.57 and dissolved oxygen 5.125 to 5.205 mg/L during a 40-day feeding comparison with weekly siphoning
Endpoints: survival, absolute length gain, absolute weight gain, ordinal visible-red colour score, feed proximate composition, temperature, pH, dissolved oxygen.
What it can support: Across the reported 40-day treatment means, Tubifex aquaria had the highest survival mean at 85.00 percent and highest colour-score mean at 3.50. Reported length and weight gains were not statistically distinguished among feeds. The comparison supports only that these four complete feed treatments produced different reported survival and visible-colour patterns under this protocol.
What it cannot support: The animals were identified only to genus, and source line, sex, age, life stage, starting size, starting colour balance and randomization method were not reported. Feed amounts and actual intake were not measured, the live foods and pellet differed in physical form and proximate composition, the Spirulina pellet was not identified, and proximate values lack replicate error and a stated wet- or dry-matter basis. Aquarium is the assigned unit, but the analysis does not state whether aquarium means were modeled, how many animals entered each endpoint or whether colour scoring was blinded under standardized lighting and background. The written method describes four colour scores while Figure 1 shows five with conflicting descriptions. The table note says different letters indicate a difference at P greater than 0.05 while the text says P less than 0.05. The discussion substitutes Hermetia illucens for Daphnia in one passage and later says Chironomus and Daphnia produced more intense colour than Tubifex, contradicting the table. Water ranges are pooled rather than treatment-specific, and nitrogen, mortality timing, feed-borne organisms, disease, reproduction, composition and long-term safety were not measured. This does not establish Tubifex as a complete or safest food, a live-feed dose, a colour-grade method or a recommendation to introduce wild or cultured worms into a home aquarium.
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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.
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CC BY quantitative and image dataset
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Field observation
Direct Neocaridina evidence
Prati et al., 2025. BioInvasions Records 14(2): 403-419.
Source taxon as published: Neocaridina davidi; Ecytonucleospora hepatopenaei, Orthosomella lipae, Microsporidium sp. SRE01 and Microsporidium sp. SRE02 as named by the source
Study shape: Forty shrimp were dip-netted at each of three sites and preserved immediately in 96 percent ethanol. All were identified morphologically, measured, sexed or classified as immature, dissected, visually screened internally and tested molecularly. Host CO1 and microsporidian SSU rRNA amplicons were Sanger sequenced and placed with reference sequences in maximum-likelihood phylogenies with 1000 bootstrap replicates.
Experimental unit: Individual field-collected shrimp for morphology, internal visual screening and molecular detection; one date and three sites in two catchments for the field occurrence observations
Environment: Three freshwater sites in two catchments on La Reunion Island sampled on 15 April 2024 at 24.5 to 26.2 C and 76 to 268 microS/cm conductivity
Endpoints: host morphology and CO1 identity, sex and life-stage counts, carapace length, microsporidian PCR-positive count, sequence similarity and phylogenetic placement, site occurrence, one-day temperature and conductivity.
What it can support: Five of 120 sampled shrimp were PCR positive for four reported microsporidian sequence types: two Orthosomella lipae detections at the Salazie creek, and one Ecytonucleospora hepatopenaei, one Microsporidium sp. SRE01 and one Microsporidium sp. SRE02 detection at Bras Citronnier. None of the 40 Ravine Seche shrimp was positive. All 120 hosts shared one CO1 haplotype reported as 100 percent similar to the N. davidi reference set.
What it cannot support: This is a one-day field sample from three sites, not an aquarium or trade prevalence estimate. The paper reports no histology, tissue lesions, clinical signs, survival, experimental transmission, pathogenicity or treatment endpoint, and it did not test native co-occurring shrimp for parasites. Ethanol preservation prevented epibiont analysis. The single host CO1 haplotype cannot prove a single introduction, seller line, release event or movement route between catchments. Sequence similarity cannot establish parasite origin or that N. davidi introduced any organism. One Table 2 subgroup is internally inconsistent, listing 16 immatures while reporting two positives as 2 of 14, so subgroup percentages should not be repeated as settled denominators. The one-day site temperature and conductivity readings are not aquarium care limits, and zero of 40 at one site is not proof of absence.
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Controlled experiment
Adjacent biological evidence
Tiwari et al., 2025. Integrative and Comparative Biology 65(6): 1489-1497.
Source taxon as published: Dugesia tigrina, Dugesia dorotocephala and Lumbriculus variegatus; no Neocaridina
Study shape: Starved planaria encountered injured or uninjured short and long blackworms in thirty-four recorded behavioural trials.
Experimental unit: One planarian-blackworm behavioural trial
Environment: Thirty-five-millimetre freshwater Petri-dish predation trials near 21 C
Endpoints: grip and wrap behaviour, feeding success, prey injury and length context.
What it can support: The tested planaria used adhesion during blackworm capture, and prey injury changed the observed interaction.
What it cannot support: Blackworm capture does not establish predation on Neocaridina eggs, juveniles or adults, and body shape alone cannot identify the aquarium flatworm.
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System observation or method study
Direct Neocaridina evidence
Wild et al., 2025. Animals 15(12): 1715.
Source taxon as published: Neocaridina genus, cherry-red breeder line without species-level identification
Study shape: Sixty euthanized shrimp were divided across 12 fixation and processing variants, implying five individuals per variant and confirmed as n = 5 for nucleus-area results. Neutral-buffered formalin, Bouin fluid and Davidson fluid were compared alone and after trypsin digestion, hydrochloric-acid decalcification or abdomen removal before fixation and decalcification.
Experimental unit: Individual post-mortem specimen within a five-shrimp processing variant; five slides with three sections each and 150 measured muscle nuclei per specimen were subsamples, not additional independent animals
Environment: Post-mortem laboratory histology comparison after rearing in three 24 L nursery tanks
Endpoints: section fragility and tearing, subjective fixation, autolysis and staining scores, muscle-cell nucleus area, digital RGB staining measurements, hepatopancreas, ventral nerve cord and muscle preservation.
What it can support: Histological appearance in Neocaridina specimens depends strongly on fixation and processing. Davidson fluid gave the best unmodified preservation among the three tested fixatives, while trypsin and acid decalcification could add tissue damage, autolysis or staining artifacts; abdomen removal followed by Davidson fixation and decalcification scored best overall in this laboratory comparison.
What it cannot support: This was a post-mortem laboratory method study, not a live-animal health, diagnosis, treatment or husbandry experiment. The source identified animals only to genus and used one breeder line, did not report sex, random allocation, assessor blinding, inter-rater agreement or tank-to-treatment balance, and used only five animals per variant. Multiple sections, nuclei and pixels are specimen subsamples. Subjective score sums were reported without uncertainty, and one-way ANOVA with Fisher NIR post hoc testing does not remove small-group or multiplicity concerns. Autolysis and altered staining can be preparation artifacts rather than evidence of illness before death. The chemicals and dissection steps are hazardous laboratory procedures, and the reported 40 mg/L MS222 method lacks time-to-death, confirmation and buffering details, so it is not a validated home euthanasia protocol.
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Controlled experiment
Direct Neocaridina evidence
Hou et al., 2024. PLOS ONE 19(12): e0315585.
Source taxon as published: Neocaridina davidi var. red
Study shape: Five diets differing in astaxanthin and Bidens pilosa were compared with repeated colour measurements, gene expression and a later sealed-bottle challenge.
Experimental unit: Independently filtered jar for the diet treatment and dissolved-oxygen bottle for the later challenge
Environment: Independently filtered six-litre jars at 24 C for 56 days followed by a nine-hour sealed-bottle challenge
Endpoints: CIELAB colour, colour-related gene expression, stress-related gene expression, nine-hour colour stability.
What it can support: The tested additives changed measured colour values and selected gene-expression outcomes under the reported diet and challenge conditions.
What it cannot support: It does not establish genetic colour improvement, permanent grade, general health benefit, transport success or a safe dissolved-oxygen threshold.
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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.
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CC BY quantitative and image dataset
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Field observation
Direct Neocaridina evidence
Prati et al., 2024. Biological Invasions 26: 2499-2523.
Source taxon as published: Neocaridina davidi
Study shape: Feral occurrence, environmental conditions and intracellular parasite relationships were investigated.
Experimental unit: Field population, site and sampled animal, depending on endpoint
Environment: European feral populations with field sampling and molecular parasite work
Endpoints: feral occurrence, range expansion, microsporidian detection, environmental context.
What it can support: Feral populations and intracellular parasite associations exist in the sampled locations.
What it cannot support: One site value is not a care boundary, and field parasite detection is not a prevalence estimate for the aquarium trade.
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Controlled experiment
Direct Neocaridina evidence
Rodriguez et al., 2024. Scientific Reports 14: 9536.
Source taxon as published: Neocaridina davidi
Study shape: Sequential concentration and exposure-time series assessed visible sedation, movement, image-derived heart rate and recovery in clean tank water.
Experimental unit: Individual shrimp immersion and observation; concentration series were run sequentially rather than as replicated aquaria
Environment: Individual 300 mL laboratory immersion baths using eugenol dissolved with ethanol
Endpoints: time to observed sedation, movement during sedation, image-derived heart rate, observable recovery.
What it can support: Eugenol immersion reduced movement and image-derived heart rate under the tested research protocol, with substantial individual response variation at lower concentrations.
What it cannot support: The study did not include an ethanol-only vehicle control, long-term survival or chronic sublethal follow-up. Observable recovery is not proof of no harm, and the research imaging protocol is not a home handling, treatment or euthanasia instruction.
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Morphology or taxonomy
Direct Neocaridina evidence
Hoitsy et al., 2023. Magyar Allatorvosok Lapja 145(6): 351-358.
Source taxon as published: Owner-submitted captive Neocaridina davidi from multiple breeders; host-identification method not reported
Study shape: Owners submitted 15 shrimp from different breeders after seeing moving white organisms on the day of introduction or within the following days. The shrimp were photographed, and associated live worms were collected into water drops, examined by stereomicroscopy and light microscopy, and identified from stated morphological references.
Experimental unit: Selected submitted shrimp for carrier observations and an unreported number of removed worms for morphological identification; five shrimp shared each of three holding aquaria, but no treatment or outcome comparison was assigned
Environment: Fifteen selected diagnostic submissions held five per approximately 18-litre aquarium at reported pH 6.8, conductivity 300 microsiemens and 25 C during examination
Endpoints: visible worm count and body region, worm length, live worm morphology, species identification, presence of worm eggs in the branchial chamber.
What it can support: All 15 selected shrimp carried visible opalescent-white moving worms, with 2 to 8 observed on the head, antennae or rostral region. Examined worms were reported as 0.8 to 2.0 mm long and identified as Scutariella japonica using paired anterior projections, two eyespots, internal reproductive anatomy and a posterior horseshoe- or heart-shaped sucker. Some worms contained eggs, and eggs were visible through the lateral carapace in the branchial chamber.
What it cannot support: This was a selected diagnostic case series, not a prevalence, transmission, quarantine-duration, water-quality or source-risk study. Host identity was not independently verified, source counts and clustering were not reported, the number of worms examined was omitted, and no molecular confirmation, voucher or image-accession record was stated. The study measured no respiration, gill lesion, molting failure, growth, reproduction, mortality, spontaneous clearance or treatment outcome. Harm statements were literature synthesis, not observations in these 15 shrimp. Fenbendazole was mentioned in one discussion sentence without product, dose, exposure, allocation, control, efficacy, recurrence or host-safety data. It cannot support a treatment recipe, a claim that every white worm is S. japonica or use of treatment response as diagnosis.
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Related source material:
University of Veterinary Medicine Budapest repository record and open PDF
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Controlled experiment
Direct Neocaridina evidence
Lima, 2023. UNESP undergraduate thesis, 22 pages.
Source taxon as published: Aquarium-shop Neocaridina davidi stock and aquarium-shop Biomphalaria glabrata; diagnostic methods and snail infection status were not reported
Study shape: Forty-two one-week-old shrimp were randomly selected and individually assigned as 14 animals per treatment to excess daily Sano S-PAK pellets, excised B. glabrata mantle, or an equal pellet-and-mantle offering. Animals were weighed every 14 days and sexed after 56 days.
Experimental unit: Individual 600 mL beaker containing one shrimp. The final analysis and figure contain 14 pellet, 13 snail-only and 13 mixed animals, but the loss of two of the 42 starters is not reconciled with the reported 100 percent survival.
Environment: Fifty-six-day laboratory feeding experiment at 25 C in individual 600 mL beakers with 1.6 g wet Java moss, daily partial water replacement and biofilm removal every 14 days
Endpoints: initial and final wet mass, mass gain, daily mass gain, specific growth rate, day-56 phenotypic sex, reported survival.
What it can support: Under this short individual-beaker protocol, the reported two-factor ANOVA detected no feeding-treatment main effect on mass gain and detected a sex main effect, with females gaining more mass than males.
What it cannot support: Failure to detect a treatment difference is not an equivalence or noninferiority test, and the small post-hoc sex groups were highly imbalanced. Food was offered in excess, but portion, consumed mass, leftovers, leaching and feed conversion were not measured. Snail mantle composition, pathogen screening, storage and water effects were not tested, and the snails had been raised on the same pellet used as the comparator. The abstract swaps the snail-only and mixed sex counts relative to the results, the stated 100 percent survival conflicts with 42 starters and 40 analyzed animals, and the mixed-female table mean of 0.335 g is impossible relative to the treatment final mass and plotted values. The paper reports no diet-by-sex interaction, exact treatment contrasts, power or raw data. It cannot establish that snail tissue is nutritionally complete, equivalent to formulated feed, pathogen-safe, humane to prepare, suitable as an infestation-control method or appropriate for routine home feeding.
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Morphology or taxonomy
Direct Neocaridina evidence
Maciaszek et al., 2023. Animals 13(10): 1616.
Source taxon as published: Neocaridina davidi
Study shape: One hundred shrimp from each of three locations within each source category were inspected individually in rostral, branchial, pereiopodal and pleopodal regions. Epibionts were isolated under clove-oil and ethanol sedation, identified morphologically at 500 to 1500x, and followed by an incompletely reported two-week quarantine observation. Prevalence models used source category and sex, microhabitat models used body region, and Spearman correlations assessed co-occurrence, egg production and molting.
Experimental unit: Individual examined shrimp nested within nine source locations and their transport bags; the reported individual-level GLMs do not describe source location or bag as a cluster or random effect
Environment: Nine hundred approximately 2.0 cm shrimp collected in November 2018 from three wild sites, three aquaculture-pond sources and three aquarium sources in Taiwan, then air-freighted in habitat water to Poland and inspected after a 10-hour acclimation in the transport-bag water
Endpoints: morphological identity and measurements, prevalence and mean intensity, co-occurrence, body-region distribution, egg-production association, molting association, observed cuticle traces and recolonization after molt.
What it can support: The authors distinguished Cladogonium kumaki, Monodiscus kumaki, Scutariella japonica and Holtodrilus truncatus, plus ciliates and rotifers. At least one recorded epibiont occurred on 688 of 900 shrimp and H. truncatus on 390 of 900. All four named taxa occurred in all four body regions with different distributions; H. truncatus was relatively associated with rostral and pereiopodal regions. Live H. truncatus and temnocephalids were observed leaving shed exuviae and seeking a host, sometimes recolonizing the nearby original shrimp.
What it cannot support: The source categories were observed, not assigned treatments, and are represented by only three locations each. Transport, ten-hour acclimation and microscopy handling may have changed organism abundance or location. Source-level clustering was not included in the reported GLMs, the sample was strongly female-skewed, and the sexing method was not stated. Morphological identification required microscopy and does not validate a phone-image diagnosis. Egg and molt analyses were correlational; the H. truncatus molt row reports rho -0.26 with p 0.847, an internally inconsistent combination for the surrounding interpretation. The quarantine allocation, denominator, water conditions, removal exposure and outcome measurements were not fully reported. Molting did not clear live worms, and the paper did not assign or compare a salt, medication or home-removal treatment. The sample is not prevalence for every seller, home colony or geographic trade route.
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Controlled experiment
Direct Neocaridina evidence
Melo, 2023. Doctoral thesis, Federal University of Minas Gerais.
Source taxon as published: Neocaridina davidi
Study shape: Separate total-ammonia and nitrite experiments used a control plus five treatments, two vessels per treatment and ten juveniles per vessel.
Experimental unit: Exposure vessel, not each shrimp
Environment: Renewed 96-hour acute laboratory exposures at about 25 C
Endpoints: 96-hour mortality, median lethal estimates, gill histology.
What it can support: Direct acute juvenile ammonia and nitrite exposure-response estimates with stated nitrogen reporting bases.
What it cannot support: A doctoral thesis with limited treatment replication does not provide chronic nitrate, chronic no-effect levels, transport limits or home targets.
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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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Morphology or taxonomy
Neocaridina identity evidence
Kakui and Komai, 2022. Aquatic Animals 2022: AA2022-1.
Source taxon as published: Neocaridina sp. aff. davidi
Study shape: Associated worms and host shrimp were examined for a regional record and taxonomic identification.
Experimental unit: Collected host or associated worm specimen
Environment: Field-collected host and associated worm specimens from Hokkaido, Japan
Endpoints: worm identification, host association, regional occurrence.
What it can support: A method-based record of Scutariella japonica associated with the source host identification.
What it cannot support: Occurrence does not establish aquarium prevalence, severity, treatment efficacy or identification from a distant white shape.
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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
Santoso et al., 2022. Clarias: Jurnal Perikanan Air Tawar 3(1): 18-27.
Source taxon as published: Neocaridina heteropoda
Study shape: Five transport durations of 24, 48, 72, 96 and 120 hours were assigned in a completely randomized design with three bag replicates per duration. Bags were rocked every two hours. Arrival survivors were then observed for three days, but the holding vessel, allocation, feeding and transfer methods were not described.
Experimental unit: Transport bag for the arrival comparison, with three bags per duration. The article does not report shrimp per bag or identify the independent holding unit for the survivor-conditioned three-day follow-up.
Environment: Simulated closed transport in 15 by 30 cm conventional plastic bags with 125 mL settled well water, Hydrilla shelter, insulated packing and 15 minutes of rocking every two hours
Endpoints: arrival survival, three-day survival conditional on arrival survival, temperature and pH before transport, after transport and after holding, dissolved oxygen before transport, after transport and after holding, carbon dioxide after transport, test-kit result labelled ammonia or NH3 after transport.
What it can support: Under this incompletely specified conventional-bag package and rocking simulation, arrival survival differed across duration groups and declined from 96 percent at 24 hours to 54.17 percent at 120 hours. Reported three-day survival was conditional on having arrived alive and did not differ statistically among duration histories.
What it cannot support: Only three bags represented each duration, no zero-hour control was included, shrimp per bag, gas volume, oxygen-fill procedure and gas-to-water ratio were not reported, and source identity was not independently verified. The percentage increments are not permission to infer an omitted bag count. The study used arcsine-transformed survival with Kruskal-Wallis and pairwise Mann-Whitney tests but did not report exact test statistics or a multiplicity correction. Follow-up vessel count, volume, allocation, feeding, water exchange and transfer method were absent, so conditional survivor percentages are not replicated delayed-survival evidence or recovery to the original cohort. Carbon dioxide and the result labelled ammonia or NH3 were tabled only after transport, with no validated analyte basis, time series or toxicity analysis. The conventional package does not validate a breather bag, package density, shipping weather gate, ammonia threshold, acclimation rate, prompt-transfer protocol or acceptable maximum duration.
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Field observation
Direct Neocaridina evidence
Schneider et al., 2022. Diseases of Aquatic Organisms 150: 125-130.
Source taxon as published: Neocaridina davidi; Enterocytozoon hepatopenaei and Microsporidium sp. EFB01 as named by the source
Study shape: Seventy-five ethanol-preserved adults were identified morphologically and screened for microsporidians by PCR from muscle tissue. Three randomly selected hosts received CO1 sequencing. Positive microsporidian amplicons were sequenced, compared with GenBank and placed in maximum-likelihood trees with 999 bootstrap replicates.
Experimental unit: Individual field-collected shrimp for PCR detection and carapace length; three randomly selected shrimp for host CO1 confirmation; one stream, one site and one sampling date for the population observation
Environment: One dip-net collection on 14 December 2021 from the Finkelbach, a German tributary of the River Erft, at 9 C, pH 7.65, dissolved oxygen 9.8 mg/L and reported conductivity 620 microS/cm
Endpoints: host identification, PCR-positive microsporidian count, sequence similarity, phylogenetic placement, sex, carapace length, visible epibiont and parasite examination.
What it can support: Four of 75 sampled shrimp were PCR positive for microsporidians. Three yielded sequences reported as 99.56 percent similar to Enterocytozoon hepatopenaei isolates, although only two were suitable for phylogenetic analysis; the fourth yielded the unresolved Microsporidium sp. EFB01 sequence. The three host CO1 sequences were 99.68 percent similar to one Taiwanese N. davidi reference.
What it cannot support: This is a one-date field detection from one German stream, not an aquarium-trade prevalence estimate or a transmission study. Only three hosts received molecular confirmation, and the paper does not report extraction blanks, PCR negative controls, product lengths, histology, tissue lesions, clinical signs, survival, transmission tests or treatment. PCR from muscle tissue cannot establish visible diagnostic characters or disease severity. The three E. hepatopenaei-positive shrimp were smaller than the 71 uninfected shrimp, but no supported inferential comparison was reported and three positives cannot establish growth suppression. Sequence similarity does not establish where either organism originated, that N. davidi introduced it, pathogenicity to native fauna, zoonotic risk or a home treatment.
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Field observation
Direct Neocaridina evidence
Bauer et al., 2021. Diseases of Aquatic Organisms 146: 107-115.
Source taxon as published: Neocaridina davidi
Study shape: Twenty submitted shrimp were assessed with clinical examination, wet mounts, microbiology, histology and sequencing of green pleopodal structures.
Experimental unit: Submitted shrimp, tissue section or algal sample, depending on method
Environment: Diagnostic examination of an imported commercial group during a multifactorial outbreak
Endpoints: organism morphology, sequence placement, cuticle penetration, co-occurring microbes, outbreak mortality history.
What it can support: The green organism was placed near Trentepohliales and rhizoids were observed penetrating cuticle into subcutaneous tissue.
What it cannot support: The multifactorial case does not assign every death to the alga, establish prevalence or let a green phone image confirm the organism.
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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
Ismandar, Dewantoro and Rachimi, 2020. Borneo Akuatika 2(1): 1-7.
Source taxon as published: Neocaridina denticulata sinensis
Study shape: Four target-water cooling treatments of 13, 15, 17 and 19 C were compared with three replicate groups per treatment. The report says temperature was lowered gradually to the target over or for 10 minutes, animals were packed without water at a nominal 17 C package temperature, and the insulated box traveled for 24 hours in a closed vehicle.
Experimental unit: Replicate transport group within a temperature treatment. The methods report 120 shrimp total, four treatments, three replicates and parenthetically 10 shrimp per treatment; those figures are compatible only if 10 means per replicate, so the written animal-unit description is internally inconsistent.
Environment: Low-temperature dry transport in a 30 by 30 by 40 cm insulated box layered with ice, Imperata grass, shrimp, grass and ice after gradual cooling; the package was intended to remain at 17 C
Endpoints: survival immediately after 24-hour transport, time from normal behaviour to reported immobility, time from reported immobility to movement or consciousness response, cooling behaviour, group mean weight change.
What it can support: The results table reports immediate post-transport survival of 86.67 +/- 5.77, 76.67 +/- 11.55, 50.00 +/- 10.00 and 33.33 +/- 5.77 percent for the 13, 15, 17 and 19 C target groups. The same groups had reported induction times of 203, 341, 431 and 530 seconds and reported recovery-response times of 3, 5, 28 and 33 minutes. Survival at 13 and 15 C was not statistically different in the article.
What it cannot support: This atypical dry chilled protocol is not a conventional or gas-permeable water bag and does not validate home cooling or anesthesia. The abstract mislabels treatment B as 13 C while giving the 15 C result, the results table gives higher survival and lower weight change at 13 C, and the conclusion calls 15 C best without resolving that conflict. The text also reports panic-like behaviour at 15 to 17 C while the conclusion says gradual cooling avoided a panic phase. The cooling wording does not distinguish ramp duration from a target-temperature hold, grass moisture and conditioning were not specified, and no package-temperature trace, detailed checks, recovery-water method, recovery denominator or delayed survival was reported. The weight-change formula uses grams while the table uses milligrams, the abstract reports 0.1 mg while the body reports 0.01 mg for 13 C, and recovery time was not included in the stated ANOVA plan even though the conclusion calls it significantly affected. Loss of balance, weak response and immobility were experimental endpoints, not normal receiving signs or permission to reproduce the protocol.
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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
Włodarczyk et al., 2019. PeerJ 7: e7399.
Source taxon as published: Adult Neocaridina davidi obtained from local shrimp breeders
Study shape: Non-starved controls were compared with adults deprived of feed for 14 days and with groups re-fed for 4, 7 or 14 days after the same deprivation. DHE flow cytometry and confocal imaging assessed reactive-oxygen signals, immunofluorescence assessed MnSOD and a control-tissue western blot checked antibody specificity.
Experimental unit: Shrimp were isolated individually, but Table 1 lists 24 animals per flow-cytometry group and four per confocal method while the statistical section says assays used 5 to 6 samples in duplicate; the animal-to-assay sample mapping and any pooling are not fully reported
Environment: Adults were held in one 40-litre laboratory tank at 21 C, pH 7 and 10 dH, then isolated in shaded 250 mL containers with algae suppressed, excrement removed and 10 percent of the water replaced daily
Endpoints: percentage of DHE-labelled ROS-positive cells, qualitative tissue DHE signal, qualitative MnSOD immunofluorescence, antibody-specificity western blot.
What it can support: Under the tested algae-suppressed conditions, 14 days without feed increased the reported mean ROS-positive cell fraction from 2.8 to 13.2 percent in the hepatopancreas and from 1.3 to 12.7 percent in the intestine. Reported values declined toward control levels after 7 to 14 days of re-feeding, alongside weaker MnSOD immunofluorescence.
What it cannot support: The study tested one 14-day deprivation, not a weekly fast, and measured dissected-cell oxidative-stress markers rather than a survival, growth, molting, behaviour, reproduction or longevity benefit. Its isolated 250 mL containers deliberately suppressed algae and do not represent a conditioned community aquarium. Random allocation and blinding were not reported, sex was omitted from the analysis, assay sample mapping is incomplete, MnSOD treatment differences were not quantitatively measured, and the table, figure and statistical-method descriptions make the comparison-letter interpretation ambiguous. It cannot establish that routine fasting is beneficial, that missing one normal feeding is harmful, that biofilm is adequate, or that a specific re-feeding schedule is safe.
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Controlled experiment
Direct Neocaridina evidence
Włodarczyk, Student and Rost-Roszkowska, 2019. Canadian Journal of Zoology 97(4): 294-303.
Source taxon as published: Adult male and female Neocaridina davidi obtained from local shrimp breeders
Study shape: Adults were deprived of feed for 7, 14 or 21 days, and subsets were re-fed for 4, 7 or 14 days. TEM, TUNEL, LysoTracker and caspase-3 methods assessed intestine and hepatopancreas cells. Non-starved reference values and images were imported from a 2016 paper rather than produced as a concurrent control group.
Experimental unit: Shrimp were isolated individually, but Table 1 lists 10 TEM animals and 8 animals for each confocal method in most groups while quantitative Tables 2 and 3 report n = 5; animal reuse, pooling and the cell-to-animal analysis hierarchy are not resolved
Environment: Adults came from one 40-litre laboratory tank at 21 C, pH 7 and printed total hardness of 10 degrees d, then were isolated in shaded 250 mL containers with algae suppressed, excrement and exuviae removed and 10 percent of the water replaced daily
Endpoints: midgut cell ultrastructure, autophagosome-bearing cell percentage, TUNEL-labelled cell percentage, qualitative LysoTracker signal, qualitative caspase-3 signal, unquantified mortality during 21-day deprivation.
What it can support: Within this linked laboratory series, 7 and 14 days without feed were associated with more reported autophagosome-bearing midgut cells, and tissue descriptions after 14 days included fewer organelles and less reserve material. After re-feeding, qualitative tissue structure and the reported cellular markers moved toward the earlier non-starved reference.
What it cannot support: This paper reused the prior experimental procedure and imported its non-starved controls, so it is not independent replication and does not provide a concurrent baseline. Most animals deprived for 21 days reportedly died, but mortality counts, timing and survival analysis were absent; the paper says that group was not quantitatively analyzed even though Table 1 lists post-21-day TEM specimens. Re-feeding food amount, frequency and intake were not reported. Table 1 animal counts conflict with the n = 5 quantitative tables, the nested cell and animal hierarchy is unclear, random allocation and blinding were not reported, and the t tests omit comparison pairs, assumptions, exact p values and multiplicity handling. Table 3 prints zero means with positive standard deviations for nonnegative percentages, which is internally impossible. Caspase-3 and LysoTracker results were qualitative, marker validation was limited, and the TUNEL threshold was described as arbitrary. The study measured no growth, molting, behaviour, reproduction, longevity or whole-animal recovery and cannot establish a routine fast, safe missed-feeding interval, re-feeding recipe, home diagnosis or health benefit.
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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
Marques, 2018. Integrated MSc dissertation in Veterinary Medicine, University of Lisbon.
Source taxon as published: Commercial adult shrimp labelled Neocaridina davidi var red; no morphological or molecular identification method was reported
Study shape: The first four-week trial placed a new 15-shrimp group in each of four aquaria each week while one lamp colour remained assigned to each aquarium. The second 20-day trial kept four 15-shrimp groups in their aquaria while rotating the four lamps every five days. An observer counted shrimp inside the lit Petri dish, inside the unlit Petri dish and elsewhere every five minutes during five one-hour periods per day. Lamp spectra were reported as relative spectral distributions, and the lit-dish count was analysed with a generalized linear mixed model followed by Tukey comparisons.
Experimental unit: In the first trial, aquarium and light colour were completely confounded even though four successive shrimp groups entered each tank. In the second, aquarium-period was the assigned unit, giving four five-day tank-periods per colour while the same four groups were counted repeatedly. The thesis reports 5,200 count occasions per trial but does not state the model formula, response family, random effects, repeated-measures correlation, lamp order or carryover treatment.
Environment: Four 15-litre aquaria at a reported 23 C, pH 7 and 60 mg/L GH, each holding 15 adults and two Petri dishes of black stones, with only the left dish illuminated by a white, blue, red or green Marina LED
Endpoints: number inside the lit Petri dish, number inside the unlit Petri dish, number elsewhere in the aquarium, qualitative movement, observed mortality, births and molts.
What it can support: Under this two-dish apparatus, the mean number counted inside the lit dish differed by lamp colour in both trials. Reported lit-dish means were 3.70 red, 1.76 green, 0.93 white and 0.03 blue in the first trial, and 4.03 red, 1.74 green, 1.32 white and 0.01 blue in the rotating-lamp trial. The result supports a short-term colour-associated location-count signal that merits a better controlled replication.
What it cannot support: The thesis does not establish a preferred aquarium spectrum or a welfare ranking. It compared one coloured lit area with one dark area rather than offering simultaneous colour choices, kept the lit area on the left, did not measure or equalize irradiance or illuminance, reported only normalized spectral shapes and did not report observer blinding. Red shrimp can also differ in detectability under different coloured lamps. The stated 20:00 to 08:00 light period does not overlap the stated 09:00 to 19:00 observations, leaving the exposure timing internally unresolved. The first-trial methods state 240 adults while the abstract states n = 120. Phase one cannot separate lamp colour from aquarium; phase two does not report lamp sequence, washout or carryover control; and the statistical formula and random effects are absent despite thousands of correlated repeat counts. Sex, individual identity, lamp output drift and births entering counts were not resolved. No stress biomarker, growth, survival comparison, reproductive rate or chronic colony outcome was measured. Absence of observed deaths and incidental births or molts cannot show comfort, health benefit or reproductive superiority. Do not recommend red light, warn against blue light, specify a spectrum or photoperiod, infer colour vision, or use lamp attraction as a capture method from this study.
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Controlled experiment
Direct Neocaridina evidence
Sonakowska et al., 2017. PLOS ONE 12(3): e0173563.
Source taxon as published: Neocaridina davidi
Study shape: Midgut cell and mitochondrial responses were compared through starvation and refeeding periods.
Experimental unit: Animal and prepared midgut tissue
Environment: Laboratory starvation and refeeding experiment with tissue analysis
Endpoints: cell structure, mitochondrial potential, regeneration after refeeding.
What it can support: Starvation and refeeding changed method-confirmed midgut cell outcomes.
What it cannot support: It does not establish a routine fast, nutritional adequacy, a safe missed-feeding duration or a diagnosis from appearance.
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Morphology or taxonomy
Direct Neocaridina evidence
Ahn and Min, 2016. Journal of Species Research 5(3): 459-462.
Source taxon as published: Wild stream shrimp identified only as Neocaridina sp.; no host voucher, diagnostic method or molecular host identification reported
Study shape: The authors removed worms from preserved host exoskeletons under a stereomicroscope, described external and internal morphology, deposited examined material at Inha University and the National Institute of Biological Resources, and extracted whole-body DNA from one worm for partial mitochondrial COI amplification and sequencing.
Experimental unit: An unreported number of Neocaridina hosts and removed worms across three collection dates; one whole worm for the 674-base-pair COI sequence; three National Institute of Biological Resources voucher identifiers were listed
Environment: Hand-net collections from Andeok Valley, Jeju-do Province, South Korea on 23 June 2010, 23 August 2015 and 18 May 2016; hosts were preserved directly in 95 percent ethanol at the collection site
Endpoints: geographic and host occurrence, whole-body and jaw morphology, body length, voucher deposition, partial mitochondrial COI sequence.
What it can support: The record establishes morphologically identified H. truncatus on wild Neocaridina sp. at one South Korean stream across material dated in 2010, 2015 and 2016. Described characters included a transparent terete body under 2 mm, no trunk appendages, a posterior sucker about the diameter of the head region and paired jaws with a 7/7 dental formula. Three NIBR voucher identifiers and the first reported 674-base-pair COI sequence for the genus, GenBank KX683299, provide checkable taxonomic anchors.
What it cannot support: The paper does not state the number of hosts collected, hosts carrying worms, worms examined per date, worms per host, sampling effort or whether absence was assessed, so three dated collections do not estimate prevalence, persistence between dates or geographic range. The host remained Neocaridina sp. without a reported voucher or molecular identification. Although examined material was deposited at two institutions, only three NIBR identifiers were listed and the paper does not map individual morphology, image, sequence and voucher records. The single COI sequence created a barcode reference but was not compared with an existing conspecific sequence or used in a phylogenetic or species-delimitation analysis. Immediate ethanol preservation precluded live attachment, movement and host-response observations. The study measured no lesion, respiration, molting, growth, reproduction, mortality, transmission, origin, treatment, recurrence or host safety. It cannot establish N. davidi prevalence, aquarium-trade origin, harmlessness, a phone-image diagnosis, a quarantine duration or a treatment protocol.
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Related source material:
Korea Citation Index open article record and licensed PDF
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Controlled experiment
Adjacent biological evidence
Rivera-De la Parra, Sarma and Nandini, 2016. Journal of Limnology 75(s1).
Source taxon as published: Hydra predators and cladoceran, copepod and rotifer prey; no Neocaridina
Study shape: Hydra predation was compared across named zooplankton prey, and Hydra-conditioned water was tested against Daphnia demography.
Experimental unit: Predator-prey culture vessel
Environment: Laboratory freshwater predator-prey cultures
Endpoints: prey capture, prey susceptibility, Daphnia population growth.
What it can support: The tested Hydra captured several zooplankton taxa and affected one Daphnia population endpoint under the laboratory conditions.
What it cannot support: No Neocaridina life stage was tested, so the study does not prove that aquarium Hydra kill shrimplets or quantify colony risk.
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Morphology or taxonomy
Direct Neocaridina evidence
Sonakowska et al., 2016. PLOS ONE 11(2): e0147582.
Source taxon as published: Neocaridina heteropoda obtained from local breeders; no host-identification method reported
Study shape: Light, transmission-electron and confocal microscopy, acid-phosphatase and LysoTracker staining, TUNEL labeling and JC-1 flow cytometry were used to describe autophagy, apoptosis, necrosis and mitochondrial potential in intestine and hepatopancreas tissue from method-specific adult groups. No external stressor or disease treatment was assigned.
Experimental unit: Prepared tissue or cell suspension from method-specific adult specimens: 15 females and 15 males for general microscopy, three adults for quantitative necrosis and autophagy cell counts, two for TEM acid-phosphatase staining, 10 females and 10 males for cryosections, five pooled within each organ suspension preparation and seven for JC-1 flow cytometry; the TUNEL animal denominator was not stated
Environment: Adults maintained in 30 L laboratory aquaria at 24 C, pH 7 and 15 degrees German total hardness and fed a commercial freshwater-shrimp food
Endpoints: cell-type and organ location, autophagic structures, TUNEL-positive apoptotic cells, necrotic-cell morphology, mitochondrial membrane potential, sex comparison.
What it can support: Under the described laboratory history and without an assigned external stressor, the examined adult tissues contained autophagic, apoptotic and occasional necrotic cells. Autophagy was observed in mature intestine D cells and hepatopancreas B and F cells, apoptosis in anterior-intestine D cells and proximal hepatopancreas B and F cells, while regenerative cells showed none of the three reported processes. The study did not report a statistically significant intestine-versus-hepatopancreas difference in cells with depolarized mitochondria.
What it cannot support: This is destructive cellular and tissue evidence, not a visible home diagnosis or a disease experiment. Breeder source, sex and adult status were reported, but host identification, age, molt stage, reproductive state, tank count, animals per tank, acclimation duration, health screening, water-test methods and the TUNEL animal denominator were not. Method groups differed and their overlap was not reported. Cell counts were nested within only three adults for the necrosis and autophagy percentages, TUNEL used an arbitrary fluorescence threshold and sex-specific values or tests were not shown. Table 1 prints 938 autophagic cells from 231 total for intestine specimen 3 even though the displayed 40.26 percent implies 93. The results do not establish a gross sign, pathogen, cause of illness, welfare state, treatment target, aquarium optimum or safe exposure, and the maintained 24 C, pH 7 and 15 degree hardness history was not an assigned comparison.
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System observation or method study
Direct Neocaridina evidence
Tanaka, Wada and Hamasaki, 2016. Zoological Science 33(2): 154-161.
Source taxon as published: Holtodrilus truncatus; field hosts included Paratya compressa, Caridina leucosticta, Caridina multidentata, Caridina typus and Neocaridina denticulata
Study shape: The field survey standardized dip-net sampling at ten waterside areas per site and repeated one river seasonally. The salinity study exposed 24, 20, 30, 25 and 12 host-removed worms to nominal 0, 0.5, 1, 1.5 and 3 percent salinity without food, with observation intervals that shortened as salinity rose. Host-choice tests placed one worm with a size-matched pair of Paratya compressa, Caridina leucosticta or Caridina multidentata for 40, 36 and 39 pairwise replicates.
Experimental unit: Field host shrimp for occurrence endpoints; an incompletely reported small plastic tank holding two or five isolated worms for salinity exposure; and one worm with one two-shrimp species pair for each host-choice replicate
Environment: Field collections at one to eight sites in each of 26 rivers on the Kii Peninsula, Japan, plus 22 to 27 C freshwater laboratory salinity and two-host choice tests
Endpoints: river and host occurrence, prevalence and intensity on Paratya compressa, host body location, Paratya compressa egg number association, isolated-worm attachment-defined survival, selected host in pairwise choice.
What it can support: Holtodrilus truncatus occurred in 10 of 26 surveyed rivers and on five atyid host species. The only two examined N. denticulata were both recorded with the worm, which establishes occurrence but not prevalence. Isolated-worm survival declined with increasing nominal salinity, including significantly lower survival at 0.5 than 0 percent. In pairwise tests that excluded Neocaridina, worms selected P. compressa over both Caridina species and C. leucosticta over C. multidentata. Cohabitation was not associated with egg number in the measured P. compressa model.
What it cannot support: The direct Neocaridina result is two field animals, not a representative sample. No Neocaridina entered the salinity, host-choice or egg-number analyses. Worms were removed after the host carapace was detached, so salinity was tested on isolated organisms rather than attached worms or living host shrimp. The work measured no treatment efficacy on a host, shrimp survival, sublethal host effect, recurrence or safety margin. Worms shared exposure containers in groups of two or five, the number of independent tanks was not reported, checking frequency differed by concentration, no food was provided, and floating or failure to attach was classified as death. Nominal salinity tolerance cannot validate a salt dip, 30-second exposure, host-safe dose or Neocaridina host preference.
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Controlled experiment
Direct Neocaridina evidence
Weber and Traunspurger, 2016. Limnologica 59: 155-161.
Source taxon as published: Neocaridina davidi
Study shape: Shrimp and comparison microcosms were sampled repeatedly, followed by endpoint stomach-content examination of 40 shrimp.
Experimental unit: Model ecosystem for community effects and individual stomach for content observations
Environment: Forty-two-day freshwater model ecosystems with repeated community sampling
Endpoints: meiofaunal density, biomass, secondary production, community structure, stomach contents.
What it can support: Shrimp changed measured meiofaunal assemblages, and stomachs contained detritus, algae and several meiofaunal groups.
What it cannot support: The study did not test fish feces, aquarium sanitation, nutrient export, named pest danger or a whole-tank algae-clearance rate.
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System observation or method study
Direct Neocaridina evidence
Cheung et al., 2015. Marine Biotechnology 17(6): 811-819.
Source taxon as published: Laboratory red-patched Neocaridina denticulata denticulata linked to the Kenny et al. stock; the commercial foundation stock had no reported voucher, morphology or molecular host-identification method
Study shape: Three adult females at each of three visually assigned ovarian stages were sampled cross-sectionally. Foregut, intestine including midgut and hindgut, and hepatopancreas were dissected from every female, producing 27 individually sequenced 16S V1-V3 libraries. PCR reactions were run in triplicate and pooled within each library. Ion Torrent reads were processed in QIIME 1.8, clustered into 97 percent OTUs, assigned against Greengenes at 0.5 confidence, rarefied to 14,000 reads and compared with alpha-diversity indices, UniFrac PCoA and UPGMA trees.
Experimental unit: Individual female for ovarian-stage comparisons, with three females per stage and three linked tissue libraries per female. Triplicate PCR reactions were technical replicates pooled within a library, not additional animals. The paper did not report a repeated-measures model for tissues from the same female.
Environment: Adult females from a multigeneration recirculating freshwater laboratory stock at approximately 25 C; all were starved for two days before dissection
Endpoints: quality-filtered 16S read count, 97 percent OTU richness, Chao1 richness, Shannon diversity, phylogenetic diversity, weighted and unweighted UniFrac clustering, relative read abundance by bacterial taxon, SRA accession SRR1735538.
What it can support: Within this nine-female stock after two days of starvation, early-stage foregut and intestine libraries had lower reported alpha diversity than later-stage libraries, hepatopancreas libraries generally differed from gut libraries, and more than 80 percent of early-stage gut reads were assigned to one Coxiella-type OTU. The study provides a public 16S survey associated with tissue region and visually assigned ovarian stage.
What it cannot support: This was a small cross-sectional association, not the same females followed through ovarian development and not an assigned microbiota or reproductive treatment. Stage could be confounded with age, time, individual and egg-carrying status. Relative read abundance is compositional and does not measure absolute bacterial load, prevalence among shrimp or colonization. A short 16S Coxiella-type assignment at a low classification threshold is not a species or strain identification, Coxiella burnetii diagnosis, cultured isolate, pathogen finding or probiotic. The paper reported t tests without specifying exact contrasts, pairing, tails, variance handling, test statistics, degrees of freedom or multiplicity correction, and reported no formal group-level beta-diversity test. No extraction blank, PCR negative control, mock community, random selection or blinding was described. The paper did not test bacterial function, microbial transfer, probiotic or antibiotic exposure, fertility, spawning, hatch, offspring, growth, survival, disease, immune response, feed treatment or normal fed-tank baseline. It cannot show that microbes cause ovarian maturation, that maturation causes the microbiota difference or that changing aquarium bacteria improves breeding or health.
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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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Review or synthesis
Broader ornamental-shrimp review
Mykles and Hui, 2015. Integrative and Comparative Biology 55(5): 891-897.
Source taxon as published: Policy review comparing Neocaridina denticulata with the crayfish Procambarus clarkii and Cherax quadricarinatus, with additional inferences from other crustacean models
Study shape: The authors defined desired traits for a decapod model, compared three freshwater candidates, summarized selected Neocaridina life-history, culture, anatomy and genomic claims from earlier publications, recommended N. denticulata, and proposed obtaining a complete genome and developing tissue-specific gene-control and mutant-screening methods.
Experimental unit: Published source or narrative claim; the paper reported no new animals, aquaria, assigned treatments, measurements or statistical analysis and did not describe a systematic search, eligibility criteria or risk-of-bias assessment.
Environment: No new animal experiment or culture trial; narrative synthesis and recommendation arising from a 2015 comparative Pancrustacea symposium workshop
Endpoints: model-organism selection criteria, comparative recommendation, summary of cited life-history and culture claims, summary of draft-genome status, proposed functional-genomics next steps.
What it can support: The paper documents that two authors recommended N. denticulata as a decapod functional-genomics model in 2015 based on its small space requirement, commercial availability, transparent cuticle, cited life-history traits and then-available preliminary genome. It explicitly identified a complete genome and reliable gene-expression manipulation as future work.
What it cannot support: This policy review is not a new replication of any cited experiment and is not a systematic review. Its pH 6.5 to 8.0, temperature up to 30 C, preferred 22 to 25 C and pH 7.0 to 7.5, 15 to 16 day intermolt, 20 to 30 egg, approximately 30 day hatch and 4 to 6 month maturity statements combine older sources, populations and methods; they are not one standardized aquarium comparison or validated species-wide optimum. The broad statement that N. denticulata is resistant to bacterial infection does not establish general disease resistance, a pathogen-free line, treatment or immune supplement. The cited DPrP bacterial-challenge paper remains full-text-needed in this registry, and the companion crayfish-plague citation studied a related published species against one oomycete pathogen. The preliminary genome was explicitly incomplete, and proposed transgenesis, reporter expression, mutation screening and targeted gene control were future possibilities rather than demonstrated Neocaridina methods in this paper. The review does not validate a care range, breeding calendar, stocking rule, disease guarantee, commercial-line assay or consumer genetic intervention.
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Controlled experiment
Neocaridina identity evidence
Kenny et al., 2014. Marine Drugs 12(3): 1419-1437.
Source taxon as published: Commercially sourced red-patched animals identified as Neocaridina denticulata denticulata; no voucher, morphology or molecular host-identification method reported
Study shape: One adult supplied a 100-base-pair paired-end Illumina HiSeq2000 library with 364,013,140 reads. ABySS assembly used k-mer 51 and an assumed approximately 3-gigabase genome. A separate method experiment tested ten adults at each of five MS-222 concentrations from 1000 to 3000 mg/L and varied bath duration from 10 to 40 minutes.
Experimental unit: One commercially sourced adult for the nuclear and mitochondrial sequence resource. Individual adults were the anaesthesia units, but duration cells shown in Figure 8 contained only one to five recoverable animals and some concentration-duration combinations were omitted after insufficient induction or mortality.
Environment: Laboratory stock held in a recirculating freshwater aquarium at about 25 C; one starved and dissected adult supplied genomic DNA, while separate adults entered aerated MS-222 bath trials
Endpoints: draft contig metrics, core eukaryotic gene hits, mitochondrial genome recovery, candidate developmental gene-family hits, Daphnia protein-sequence matches, anaesthesia induction, first movement, complete recovery, three-day mortality follow-up.
What it can support: The short-read resource recovered a 15,565-base-pair mitochondrial genome and recognizable hits for 455 of 458 CEGMA core genes at the stated threshold. It also demonstrates that MS-222 concentration and bath duration changed induction and recovery under the tested laboratory method.
What it cannot support: The 99.3 percent result is a core-gene hit rate, not 99.3 percent whole-genome completeness. The draft contained 3,346,358 contigs, an N50 of 400 base pairs and 1.284 gigabases of assembled sequence against an assumed approximately 3-gigabase genome; the authors explicitly described poor recovery of non-coding regions and manually removed three bacterial-like contigs. One unvouchered seller animal cannot establish population variation, named-line ancestry, pedigree, purity, phenotype prediction or a consumer genetic test. The 3,750 putatively crustacean-specific matches depend on database content and an E-value cutoff and do not establish unique function. The anaesthesia cells were small, some animals died at 3000 mg/L and one of three died after 30 minutes at 2000 mg/L, so the protocol is not a home treatment, transport method, euthanasia instruction or general welfare guarantee.
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System observation or method study
Direct Neocaridina evidence
Niwa et al., 2014. Central European Journal of Biology 9(1): 80-85.
Source taxon as published: Wild Sugo River shrimp identified only as Neocaridina spp.; the catch could include native N. denticulata denticulata and imported Neocaridina lineages
Study shape: The authors collected 271 host shrimp across nine years, mapped H. truncatus body locations on 141 hosts in 2011, measured 23 cocoons, observed 15 host-removed worms from 14 shrimp in no-added-food survival containers, and placed five removed worms sequentially with a reported pool of 28 potential host shrimp across several taxa.
Experimental unit: Individual host for mapped attachment location; individual cocoon for diameter; nominally individual host-removed worm, although only 13 of 15 worms receive a reported container and survival range; and an unresolved sequence of five worms exposed one host at a time in the host-exchange demonstration
Environment: Field collections at three Sugo River stations from 2003 to 2011 plus laboratory observations in source river water, aerated aquaria, individual bottles, dishes and 100 mL vials
Endpoints: host body location, cocoon diameter and embryo count, observed hatching and gill-chamber entry, days survived after host removal, attachment or predation during host exchange, movement from deteriorating to surviving captive hosts.
What it can support: Among 141 mapped wild hosts, reported H. truncatus locations were 55.3 percent between the first pleopod and fifth pereiopod, 17.0 percent on the carapace, 15.6 percent at the eye base, 8.5 percent on the antennule and 3.6 percent around the egg mass. Twenty-three cocoons measured 0.58 to 0.76 mm, and the largest observed cocoon contained 14 developing worms. Ten worms in 100 mL vials survived 7 to 46 days and three in dishes survived 12 to 21 days after host removal without added food. In the host-exchange observation, all five supplied worms attached to Neocaridina hosts within three hours.
What it cannot support: Host shrimp were resolved only to Neocaridina spp. in a river where native and introduced lineages were discussed, so the findings are not species-specific N. davidi evidence. The 271 total catch, 152 animals mentioned in the attachment procedure and 141 mapped hosts are not fully reconciled, station and date denominators are missing and the location percentages do not state whether each host contributed one location or all worms were counted. No molecular confirmation of host or worm identity was reported in this article. The survival containers differed in volume, light and temperature, two of 15 worms are absent from the reported ranges, ages were unknown, water was not renewed or analytically described, death criteria were not stated and there was no fed or host-present control. The five-worm host exchange omitted exposure order, reuse mapping, independent replication, container details and complete outcomes for 28 potential hosts. Disappearance under an opaque crayfish carapace was interpreted as migration rather than directly confirmed. Claims that egg-bearing hosts hatched healthy young had no defined denominator, comparison or follow-up. Observed migration from deteriorating hosts does not establish transmission probability or a quarantine duration. The study measured no controlled shrimp lesion, respiration, growth, reproduction, mortality causation, treatment efficacy, recurrence or host safety and cannot validate predatory tankmates, forced host deterioration, worm collection, medication or a fixed isolation period.
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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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System observation or method study
Direct Neocaridina evidence
Nur and Christianus, 2013. Asian Journal of Animal and Veterinary Advances 8(1): 108-115.
Source taxon as published: Aquarium-shop animals labelled Neocaridina denticulata sinensis; identification method not reported
Study shape: About 30 females and 30 males were conditioned separately for one month. Ten selected male-female pairs then shared one breeding aquarium. Ovigerous females were moved to individual 7 by 7 by 10 cm containers for hatch observations. Thirty healthy one-week-old offspring were selected into one shared aquarium and observed through a reported life-cycle sequence.
Experimental unit: One shared breeding aquarium for the ten pairs and one shared aquarium for the 30-offspring life-cycle cohort; individual ovigerous-female containers supplied brood observations, but the methods do not explain the greater number of points visible in the brood-mass graph or whether females contributed repeated broods
Environment: Dechlorinated aerated tap water at 27 to 28 C, pH 6.5 to 7.5, dissolved oxygen above 4 mg/L, reported ammonia and nitrite below 0.1 mg/L, commercial 40 percent protein feed offered ad libitum twice daily and about 50 percent weekly water renewal
Endpoints: mating and attached-egg timing, egg length, incubation time, hatchling total length, brood count, reported juvenile and first-maturity time, female mass and brood-count relationship, water context, observed Holtodrilus occurrence.
What it can support: Under the reported culture history, selected adults mated and carried eggs, incubation was reported as 15 days at 27 C, hatchlings had a shrimp-like form and measured about 2.3 mm total length, observed broods ranged from 21 to 51, and the selected shared offspring cohort was reported to reach first maturity near 75 days and 2.3 cm total length.
What it cannot support: Species identity rested on an aquarium-shop label without a reported diagnostic method. There was one breeding tank and one 30-offspring life-cycle tank, with no independent tank replication. Only healthy one-week-old offspring were selected, and survival, losses, family contributions, observation intervals, individual tracking and the denominator reaching maturity were not reported. The brood-mass figure shows more observations than the ten-pair method explains and labels R squared as 0.3589, while the text reports 0.9587 and 0.959. Water conditions and the 40 percent protein feed were not assigned comparisons, so they are context rather than optima. High mortality during Holtodrilus occurrence lacks a denominator and causal control. The claimed 5 to 10 ppt salinity response had no assigned treatment, untreated control, dose-duration, host-safety endpoint or recurrence follow-up and cannot support a salt treatment. Study duration and season coverage were not reported, so year-round reproduction was not established. The work cannot validate a universal 15-day incubation, 75-day maturity calendar, body-size threshold, feeding recipe, water-quality target or parasite treatment.
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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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Morphology or taxonomy
Adjacent biological evidence
Van Steenkiste et al., 2013. PLOS ONE 8(3): e59917.
Source taxon as published: Dalytyphloplanida flatworms, not Neocaridina
Study shape: Complete 18S and partial 28S rDNA sequences were analysed with maximum-likelihood and Bayesian phylogenetic methods.
Experimental unit: Taxon sequence or specimen represented in the phylogeny
Environment: Molecular phylogenetic analysis across marine, freshwater and symbiotic flatworms
Endpoints: phylogenetic placement, freshwater transition reconstruction, symbiotic-lifestyle reconstruction.
What it can support: Flatworm lineages and life strategies cannot be inferred from a generic worm-like aquarium silhouette alone.
What it cannot support: The phylogeny does not identify a phone-video organism, demonstrate predation or determine whether an observed flatworm threatens shrimp.
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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
Wang et al., 2010. Israeli Journal of Aquaculture - Bamidgeh 62(2): 78-84.
Source taxon as published: Wild-collected Neocaridina heteropoda from Baiyangdian Lake; no diagnostic identification method reported
Study shape: Nine hundred shrimp were assigned at 50 per tank to six manganese-sulfate supplementation levels, with three tanks per diet. The measured complete-diet manganese concentrations were 2.79, 22.57, 42.55, 62.69, 82.59 and 102.45 micrograms per gram. Shrimp received feed at a stated daily rate of 5 percent body weight for 30 days.
Experimental unit: Tank for the six assigned diets, with three tanks per diet in one shared-water recirculating system. Figure 1 reports n = 6 for biochemical assays, but the paper does not map those samples or pooled hemolymph back to individual shrimp and tank replicates or state whether tank entered the analysis.
Environment: Thirty-day freshwater recirculating-system feeding experiment at 25 +/- 1 C and pH 8.0, with all tanks connected to one reservoir and reported water manganese of 1.00 +/- 0.02 micrograms per litre in the abstract but micrograms per gram in the methods
Endpoints: percentage length gain, percentage weight gain, hemolymph superoxide-anion assay, muscle SOD activity, muscle GPX activity, muscle CAT activity.
What it can support: Under this complete-feed formulation and 30-day system, every manganese-supplemented group had higher reported length and weight gain than the 2.79-microgram-per-gram control diet. The 62.69-microgram-per-gram measured diet had the highest reported growth percentages and antioxidant-enzyme activities and the lowest reported superoxide-anion assay value.
What it cannot support: The experiment does not establish a universal dietary requirement or deficiency threshold. The control was not manganese-free, the basal mineral premix already listed manganese sulfate, and the diets differed only within one formulation and short culture history. The paper reports neither starting-to-ending size values nor a growth formula, feed intake, feed conversion, survival, tissue manganese, deficiency signs, molting, reproduction or long-term safety. Wild-source identity, sex and life stage were not resolved. Sample-to-tank mapping for the n = 6 biochemical results and pooled hemolymph is unclear, and the water manganese unit conflicts between abstract and methods. A peak response at one tested concentration without dose-response modeling does not validate adding manganese to aquarium water, mixing a supplement into finished food or using antioxidant-enzyme activity as proof of better health.
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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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