Controlled experiment
Neocaridina identity evidence
Bai et al., 2026. BMC Genomics 27: 535.
Source taxon as published: Purchased red, yellow and blue Neocaridina denticulata strains without reported voucher, diagnostic identity method, trade names or ancestry
Study shape: Pigment-pathway homologs were identified from public BioProjects and unpublished laboratory genomic and transcriptomic data. Six adult tissues and whole juveniles were compared by 18S-normalized qPCR with three biological samples and technical triplicates. CRCN abundance was compared by western blot, and fluorescence at 380/450 nm was measured after separate cephalothorax and abdomen extraction with hepatopancreas and intestine removed.
Experimental unit: Three biological samples per strain and endpoint were reported, but the number and sex of shrimp per sample, pooling, molt state, independent source-tank history, number of culture tanks and strain-to-tank allocation were not defined; technical triplicates are subsamples
Environment: Purchased red, yellow and blue strains held in 29 x 18 x 15 cm transparent recirculation-system tanks at 26 +/- 2 C and pH 7.2 to 7.5, with the number of tanks and strain-to-tank allocation unreported
Endpoints: candidate pigment-pathway homologs and phylogenies, adult-tissue and whole-juvenile relative gene expression, CRCN western-blot band intensity relative to tubulin, body-region relative fluorescence used as a pteridine proxy.
What it can support: The tested red, yellow and blue source strains differed in selected tissue and juvenile transcript levels, CRCN western-blot abundance and body-region relative fluorescence. These cross-sectional associations identify candidate carotenoid and pteridine pathways for follow-up; the tested blue strain had higher normalized CRCN band intensity than the red and yellow strains.
What it cannot support: The study did not manipulate candidate genes, cross strains, measure inheritance, quantify a standardized visible-colour phenotype, identify or quantify individual pteridine compounds, assay carotenoid composition, assign diets or validate seller lines. Its red, yellow and blue labels were not connected to named aquarium morphs, vouchers, diagnostic species confirmation, ancestry or pedigrees. Three biological samples per endpoint, unresolved animal pooling and tank history, an unreported calibrator for 2^-delta-delta-Ct comparisons, relative fluorescence without reported tissue-mass normalization, unpublished sequence inputs and low-powered distribution tests at n = 3 limit causal or reproducible pathway ranking. The work cannot provide a genotype test, dominance chart, feed or carotenoid prescription, colour-grade prediction, ancestry claim or commercial trade-line pedigree.
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Controlled experiment
Neocaridina identity evidence
Li et al., 2026. BioTech 15(1): 15.
Source taxon as published: Neocaridina denticulata sinensis
Study shape: Expression was compared across colour strains and stages; RNA interference and sampled SNP associations were examined.
Experimental unit: Embryo group or biological sample, depending on endpoint
Environment: Laboratory expression, association and RNA-interference study
Endpoints: NinaB-like expression, red pigment deposition, SNP association.
What it can support: Functional involvement and sampled genetic associations for body-colour regulation in the source taxon.
What it cannot support: The candidate SNPs do not certify Blue Dream, Sunkist, purity, relatedness or a universal named-morph genotype.
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Controlled experiment
Direct Neocaridina evidence
Wilson, Rickward and Santostefano, 2026. Journal of Evolutionary Biology: voag042.
Source taxon as published: Neocaridina davidi
Study shape: Forty-one one-male, four-female breeding groups produced 1,191 offspring in 75 full-sibling families nested within 37 paternal half-sibling groups. Eighty shrimp from 20 families each completed six alternating control-predator assays; the other 1,111 each completed one treatment-balanced assay for pedigree-based models.
Experimental unit: Repeatedly identified individual for the 80-shrimp six-assay dataset; single individual nested within a full-sibling family and its rearing tank for the 1,111-shrimp quantitative-genetic dataset
Environment: Pet-trade-source offspring raised as separate full-sibling families in recirculating 2.9-litre tanks, then tested in five-minute open-field assays using conspecific water or water previously occupied by green swordtails
Endpoints: track length, activity above 4 cm/s, arena area covered, refuge duration, individual repeatability, additive and family variance, individual-by-environment and genotype-by-environment variation.
What it can support: The fish-water cue produced modest average decreases in track length and activity and increased refuge duration. Individuals differed repeatably, and the pedigree model attributed an estimated 24.3 percent of multivariate phenotypic variance to additive genetic effects while finding no supported variation among individuals or genotypes in cue-response plasticity.
What it cannot support: No fish entered the assay and no capture, injury, feeding, growth, survival or recruitment endpoint was measured. The cue came from one novel fish species and its strength was not an aquarium coexistence exposure. All shrimp came from one pet-trade colony, full siblings shared one rearing tank, only about two dams per sire contributed on average and additive-genetic estimates had high uncertainty. The repeated dataset always began with control, arena water was replaced after ten trials and age varied from 41 to 215 days. The study does not establish fish compatibility, a fixed personality label, a commercial-line heritability or a selection response.
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System observation or method study
Direct Neocaridina evidence
Feng et al., 2025. BMC Genomics 26: 1151.
Source taxon as published: Neocaridina denticulata sinensis
Study shape: The authors identified CRCN-family candidates in an unpublished laboratory genome, compared the family across six crustaceans, examined a 15-library five-tissue expression matrix with three libraries per tissue, ran epidermal qPCR, expressed NdCRCN-6 in Escherichia coli, generated a mouse polyclonal antibody and used western blotting and immunohistochemistry for tissue localization.
Experimental unit: Gene and genome for computational analyses; 15 transcriptome libraries with three per tissue but no reported animal-versus-pool provenance; qPCR reports three samples without defining the biological animal unit; western-blot preparations, immunohistochemistry sections and independent animal counts were not reported
Environment: Aquaculture-center shrimp 1.8 plus or minus 0.2 cm, acclimated for two weeks in a 29 by 18 by 15 cm recirculating tank at 25 plus or minus 1 C and fed Sera Shrimp Natural twice daily
Endpoints: CRCN candidate-gene count and location, gene-family phylogeny and duplication, tissue expression matrix, epidermal qPCR, antibody reactivity, epidermal and muscle immunolocalization.
What it can support: The source genome contained 24 CRCN-family candidates, mostly clustered on chromosome 23, with seven reported tandem-duplicated pairs. The 15-library matrix and qPCR showed differing tissue and epidermal expression patterns, and the generated antibody produced CRCN-family signal in epidermis and muscle under the reported assays.
What it cannot support: This is a molecular identification and localization study, not a colour-strain, diet, breeding or grading experiment. It did not measure visible colour, chromatophores, astaxanthin amount, direct astaxanthin binding in Neocaridina, growth, survival, reproduction, molt, health or selection response. One unpublished source genome cannot establish population-wide copy number, variants or commercial-line identity. Total shrimp count, tank count, sex, age, colour line, lineage and taxon-identification method were not reported. The tissue libraries do not resolve animals versus pools, and the paper does not define the biological unit behind three qPCR samples. Primer efficiencies and a calibrator were not reported for cross-gene qPCR comparisons. High CRCN sequence similarity means the polyclonal antibody may recognize multiple family members, so localization cannot assign each subunit a distinct function. Independent animals, section counts, blinded scoring and quantitative image methods were not reported. Muscle transport, epidermal colour modulation and individual-subunit functions remain inferred rather than directly measured. The accession CRA018357 is linked only to the archive homepage in the article and does not resolve these sample-provenance gaps. Twenty-four candidate genes do not mean 24 visible traits, a trade-line inheritance chart, a consumer genetic test or a colour-food recommendation.
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Controlled experiment
Neocaridina identity evidence
Huo et al., 2025. Fishes 10(3): 134.
Source taxon as published: Laboratory red, blue, yellow and wild or transparent strains labelled Neocaridina denticulata sinensis
Study shape: Adult and tissue expression used seven individual samples per strain. Embryonic expression pooled all sampled embryos from one berried female as one replicate, with seven maternal replicates per source-defined stage. For RNA interference, metanauplius embryos from each of five red-strain mothers were divided into sibling treatment and control wells of 15 embryos, exposed ex vivo after hypochlorous-acid membrane softening to 5 micrograms per millilitre NdBCO-like4 or EGFP dsRNA in ultra-pure water, and photographed after 24 hours. A transcriptome-derived candidate SNP was then targeted for genotyping in four laboratory strains.
Experimental unit: Individual shrimp for adult and tissue expression, nested within separately maintained strain tanks whose number and replication were not stated; maternal clutch for the five-mother paired RNA-interference comparison; individual shrimp for genotyping, although the reported figure totals do not reconcile with the stated 192 individuals per strain
Environment: Laboratory-owned strains selectively bred for more than three years and cultured separately in glass tanks at about 25 C, with aerated tap water, planted aquarium soil, daily compound feed and 50 percent water changes every three days
Endpoints: NdBCO-like4 expression by strain, tissue and developmental stage, RNA-interference target expression, red-channel pixel brightness ratio, pigment-particle distribution index, candidate-SNP genotype frequency, predicted RNA secondary structure.
What it can support: In the sampled laboratory strains, NdBCO-like4 expression was higher in the red strain than the yellow, blue and wild or transparent strains and varied across the five source-defined developmental stages. In the five paired maternal clutches, embryos receiving NdBCO-like4 dsRNA had lower red-channel brightness ratios and pigment-particle distribution indices than EGFP-dsRNA sibling controls under the artificial 24-hour exposure. The G.1719G>A candidate occurred in every sampled strain at different reported genotype frequencies.
What it cannot support: This is evidence of molecular involvement under the source assays, not a validated commercial-line marker, inheritance chart or consumer genetic test. The paper does not report the number of culture tanks per strain, so strain, family and shared tank history cannot be separated. The ex vivo RNA-interference protocol used removed embryos, 1 to 5 percent hypochlorous acid for 1 to 2 minutes, ultra-pure water and destructive molecular sampling; it is not a breeding or colour-enhancement method. A blank control is described but omitted from the displayed RNA-interference results, scoring regions and blinding are not reported, one reference gene was used without reported stability validation, and the split-clutch design was analysed with an independent t-test rather than an explicit paired model. The results text calls the target-expression decrease p greater than 0.05 while the discussion calls it significant. The abstract calls G.1719G>A synonymous, whereas the results and discussion call it a missense or non-synonymous D445N change. The same strain is called wild in the methods and transparent in figures. Although the methods state 192 genotyped individuals per strain, the Figure 7 genotype counts total 148 red, 95 yellow, 190 blue and 172 transparent animals, without explaining missing calls. The variant-frequency association does not establish causation, dominance, parentage, pedigree, selection response or identity for Blue Dream, Sunkist, Bloody Mary or any seller line.
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Controlled experiment
Direct Neocaridina evidence
Ishiba et al., 2025. Jxiv preprint, version 1.
Source taxon as published: Field-origin Neocaridina colony from the Tatsuta River, with five colony representatives assigned to N. davidi clade B by COI barcoding
Study shape: Sixty-four randomly selected shrimp spent seven days in a white environment, received standardized dorsal photographs, were randomly assigned by sex to four coloured environments for seven more days and were photographed again. No feed was supplied during the full 14 days. ImageJ mean RGB luminance ratios were calculated for the whole body and five body regions.
Experimental unit: Individual survivor for colour analysis; vessel count, shrimp density and animal-to-vessel mapping were not reported
Environment: Laboratory white acclimation followed by red, blue, green or white glass-sand and polyester-cord environments
Endpoints: day 14 to day 7 RGB luminance ratios, body-region RGB ratios, survival by colour environment.
What it can support: The matching RGB channel had the largest mean ratio in every environment, but only the blue group showed a significant among-channel difference, specifically red versus blue. White, red and blue environments produced some within-channel changes; green produced none. Approximate survival was 56 percent in white, 93 percent in red, 93 percent in green and 87 percent in blue.
What it cannot support: This is a version 1 preprint, not peer reviewed. Only five colony representatives were barcoded, not all 64 experimental animals. Vessel allocation and replication were not reported, so pseudoreplication cannot be excluded. Animals received no feed for 14 days; water chemistry, temperature, photoperiod, mortality timing, molt state and family contributions were omitted. The colour treatment combined sand, cord and reflected light, white survival was about 56 percent and colour results include survivors only. Repeated tests lacked a stated multiplicity correction and the analysis did not model within-animal RGB channels. The study did not test inherited colour, commercial grade, genetics, health, optimal substrate, predator detection or a camouflage survival benefit.
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Controlled experiment
Neocaridina identity evidence
Zhang et al., 2025. Animals 15(7): 901.
Source taxon as published: Laboratory red, yellow, blue and wild Neocaridina denticulata sinensis populations
Study shape: Scarb1 expression was compared across four selected colour populations and five developmental stages; five maternal clutches were split between scarb1 and EGFP dsRNA exposure, and one candidate SNP from a 216-shrimp transcriptome was targeted in four planned 384-animal population samples.
Experimental unit: Individual or biological sample for expression, maternal clutch for the paired RNAi comparison and individual shrimp for genotyping; source-population tank replication was not resolved
Environment: Laboratory culture at 25 +/- 1 C followed by removed-embryo RNA interference, expression assays and targeted genotyping
Endpoints: scarb1 expression, red pixel brightness ratio, pigment distribution scale, chromatophore development, G1593A genotype and allele frequency.
What it can support: Scarb1 knockdown changed metanauplius chromatophore measurements under the source protocol, and the synonymous G1593A genotype distribution differed sharply in the sampled yellow population.
What it cannot support: The RNAi used removed embryos from five mothers, did not report the statistical method for most comparisons and did not function at the later pre-zoea stage. The table reports 345 rather than the planned 384 genotypes per population without explaining missing calls. A selected-population association without crosses, independent line validation, phenotype prediction or a causal variant test cannot identify Sunkist or another named commercial line, establish dominance or forecast offspring.
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Morphology or taxonomy
Neocaridina identity evidence
Shih et al., 2024. Zoological Studies 63: 18.
Source taxon as published: Eight Neocaridina species or provisional lineages from Japan
Study shape: COI DNA barcoding, morphology and geographic context were combined to identify native, non-native and provisional Neocaridina lineages.
Experimental unit: Collected specimen and locality, with sequences interpreted within the sampled phylogeny
Environment: Museum-deposited specimens from eighteen sites in Kyushu and southern Honshu
Endpoints: morphological characters, COI haplotypes, species delimitation, geographic occurrence.
What it can support: Integrative evidence distinguished several co-occurring Neocaridina lineages and exposed limits of a single barcode or character.
What it cannot support: The Japanese field result does not identify a commercial colour line, establish ancestry from a photograph or resolve every competing species treatment.
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Controlled experiment
Direct Neocaridina evidence
Tropea and López Greco, 2024. Canadian Journal of Zoology 102(9): 721-734.
Source taxon as published: Neocaridina davidi
Study shape: Female reproductive timing and biochemical investment were compared with brothers, non-brothers or different male-presence conditions.
Experimental unit: Female or mating treatment unit
Environment: Laboratory relatedness and male-presence comparison
Endpoints: ovarian maturation, egg carotenoids, cholesterol, energy content.
What it can support: Relatedness changed selected reproductive timing and egg-composition outcomes under this protocol.
What it cannot support: These internal measures cannot diagnose an unseen home colony or prove commercial sources are related.
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Morphology or taxonomy
Neocaridina identity evidence
Kusmintarsih et al., 2023. Biodiversitas 24(7): 3766-3773.
Source taxon as published: Market-sourced ornamental shrimp reported as Neocaridina denticulata from Purbalingga, Central Java; no voucher-based identification was reported
Study shape: Abdominal tissue from ten visibly labelled market samples was extracted. A custom primer designed from a Neocaridina denticulata mitochondrial reference amplified an approximately 545-base-pair CO1 fragment. Bidirectional sequences were assembled, compared with one primary BLAST reference and placed in a neighbour-joining tree with selected GenBank sequences and one distant outgroup.
Experimental unit: One destructive tissue sample per visible colour entry, with ten sequence entries in the result tables; the paper does not report vouchers, specimen IDs, seller or aquarium clustering, replicate extractions, independent PCR confirmation or public accession numbers for the new sequences
Environment: Shrimp collected in August 2019 from the Purbalingga ornamental fish market and transported to a laboratory; seller, source farm, number of source aquaria and prior breeding history were not reported
Endpoints: CO1 PCR band, assembled fragment length, BLAST identity, neighbour-joining placement, reported pairwise genetic distance.
What it can support: Within this market sample and assay, the ten sequence entries labelled dark red, bright red, black, white, blue-black combinations or yellow were reported at 98.24 to 99.29 percent identity to the same Chinese N. denticulata mitochondrial reference, and the study reported zero pairwise distance among its sampled sequences. Visible colour labels therefore did not separate those sampled entries into different reported CO1 groups.
What it cannot support: This is a ten-specimen convenience sample from one market, not a survey of named global trade lines, farms, sellers or source populations. Collection wording does not resolve whether specimens came from one seller, aquarium, brood or colony. No morphological key, sex-based diagnostic structures, voucher deposition, extraction blank, PCR negative control, replicate extraction or new sequence accession is reported, so the specimen identity and sequence audit cannot be independently reconstructed from the article. The blue lane in Figure 2 has no band, the figure numbers lanes 1 to 10 while the text refers to samples 2 to 11, and Table 2 reports ten sequence entries without a standalone blue entry. The methods and Figure 3 describe a Jukes-Cantor tree, while Table 3 is labelled Kimura 2 parameter. The article reports 0.00 distance among the study entries even though their BLAST identities to the same reference vary from 98.24 to 99.29 percent, and its new sequences are not shown as deposited accessions. Low branch support appears across several reference relationships, the outgroup is a tunicate rather than a closer atyid, and the selected reference labels include disputed or inconsistent Neocaridina names. A short mitochondrial fragment records one maternally inherited locus; it cannot establish nuclear ancestry, pedigree, relatedness, genetic purity, line stability, inheritance, hybrid status or the species identity of another shrimp based on colour or a matching seller label.
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Controlled experiment
Neocaridina identity evidence
Li et al., 2022. Journal of Experimental Biology 225(6): jeb243702.
Source taxon as published: Animals sold as Neocaridina heteropoda from a local aquatic market and cultured in the laboratory for more than six months; no voucher, morphology or molecular host-identification method reported
Study shape: A tracer pilot compared 1000 ng/uL dextran with 0.05 percent Phenol Red. For editing, three Nh-scarlet sgRNAs were designed using a partial Neocaridina sequence and the Penaeus vannamei genome as reference. Cas9 at 300 ng/uL and sgRNA at 200 ng/uL were delivered at about 0.8 nL per embryo; individual sgRNAs were checked at gastrula and all three were co-injected for the flea-larva eye-phenotype screen.
Experimental unit: Individual embryo for injection and phenotype screening. Twenty-eight injected embryos survived to the reported Neocaridina screen, but the initial number injected, maternal-clutch contributions, allocation, complete control structure and tracer-table denominator or replicate basis were not reported.
Environment: One-cell embryos removed from females four hours after spawning, placed on wet filter paper with the upper surface exposed to air, injected and irrigated three times daily with sterilized freshwater through in vitro development
Endpoints: embryo survival by developmental stage, Nh-scarlet expression by developmental stage, visible eye pigmentation and shape, target-region PCR and sequencing.
What it can support: The method delivered CRISPR/Cas9 material into ex vivo Neocaridina embryos and recovered target-region insertions, replacements or deletions with altered eye phenotypes in three of 28 surviving injected embryos: two with altered eye shape and one with an absent eye.
What it cannot support: The authors called 3 of 28 surviving injected embryos a 10 percent gene-editing efficiency; the denominator excludes embryos that did not survive, and the initial injected total was not reported. Screening was based on abnormal eyes, so phenotype-normal survivors were not shown to have been systematically genotyped. Some embryos contained more than one edited sequence, consistent with mosaicism. Off-target effects were not assessed, one sgRNA lacked detectable in vitro digestion, and exact family contributions, randomization, blinding and complete negative-control comparisons were not reported. The study ended at early development and did not test adult survival, health, body colour, fertility, germline transmission, F1 inheritance, stable-line creation, commercial grade, named-trade-line identity or a consumer genetic test. The invasive embryo method is a functional-genomics protocol, not a hobby breeding procedure.
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Morphology or taxonomy
Direct Neocaridina evidence
Lu et al., 2022. Zoomorphology 141: 283-295.
Source taxon as published: Neocaridina denticulata sinensis
Study shape: The authors photographed eight source-defined embryonic stages in red, yellow and blue strains, then followed chromatophore type, form, number and distribution into early post-hatch development. Six individuals per strain were used for the detailed chromatophore observations after pigment cells appeared, and six individuals per stage per strain were dissected from metanauplius through membrane-zoea.
Experimental unit: Individual embryo or post-larva for microscopy, with six individuals per strain for detailed observations after pigment cells appeared; the paper reports one culture tank per strain, does not report family contributions to the selected eggs and does not clearly describe independence across repeated photographs or inferential comparisons
Environment: Red, yellow and blue laboratory strains cultured separately, with one strain cultured in a tank at 20 to 26 C and pH about 7.4; eggs were removed with a sodium chloride and trypsin solution, washed and cultured in a shaken 24-well plate before post-larvae were moved to a glass tank
Endpoints: chromatophore type, chromatophore form, chromatophore number, chromatophore diameter, chromatophore distribution, embryonic morphology.
What it can support: Under this in-vitro protocol, no chromatophores were reported before pre-nauplius, erythrophores were the first visible chromatophore type in all three strains at the source-defined metanauplius stage, and later stage-specific observations included leucophores in red and yellow, cyanophores in blue and xanthophores in yellow after hatching.
What it cannot support: This does not establish a universal grading age, predict adult grade from an embryo or juvenile, validate a commercial colour-line identity or reveal inheritance. One tank per strain confounds strain with tank history, family contributions were unknown and only six individuals per strain were used for detailed observations. The egg-removal, trypsin, washing, shaker and in-vitro culture protocol is not a home breeding method. Food was not assigned as a treatment, adult colour was not followed, and discussion explanations involving carotenoids, yolk or feeding relied partly on hypotheses or unpublished data rather than a diet experiment. The paper gives one-way ANOVA but does not fully report the comparison plan, repeated-measure handling, assumption checks or exact P values, and its figure and text leave some post-hatch photographic timing ambiguous.
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Controlled experiment
Direct Neocaridina evidence
Tropea, Marciano and López Greco, 2022. Zoology 151: 125990.
Source taxon as published: Neocaridina davidi
Study shape: Females were paired with brothers or non-brothers and offspring were followed through separate brood and juvenile endpoints.
Experimental unit: Mating pair or offspring culture group, depending on endpoint
Environment: Laboratory sibling and non-sibling mating comparison
Endpoints: mating, egg count, hatch count, juvenile survival, growth under feeding treatments.
What it can support: Mating and early egg measures can look similar while later hatch and juvenile outcomes differ after one full-sibling generation.
What it cannot support: It does not establish a founder minimum, multigeneration decline, colour loss, disease effect or calendar-based outcross schedule.
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Morphology or taxonomy
Neocaridina identity evidence
Zhou et al., 2021. Ecology and Evolution 11: 10502-10514.
Source taxon as published: Freshwater shrimp including Neocaridina davidi and Neocaridina denticulata
Study shape: Male morphology, COI barcodes and several molecular-delimitation methods were compared across freshwater shrimp samples.
Experimental unit: Collected specimen and sequence, nested within sampling locality
Environment: About 1,200 field specimens from forty-six sites in Henan Province
Endpoints: morphological identification, COI haplotype, molecular operational taxonomic unit, genetic distance.
What it can support: Morphological variation and molecular partitions did not reduce to one simple visual species rule in the sampled freshwater shrimp.
What it cannot support: The survey does not authenticate aquarium trade names, grades, purity or lineage, and a COI similarity threshold is not a universal species test.
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System observation or method study
Direct Neocaridina evidence
Fairfield, 2020. Doctoral thesis, University of East Anglia.
Source taxon as published: Neocaridina davidi
Study shape: Two cross-sectional analytical-method studies compared proposed molecular age markers across bounded-age aquarium cohorts. One measured global DNA methylation in 100 samples from four age groups using a commercial ELISA kit. The other sequenced amplified mitochondrial DNA from 151 samples across seven age groups, with 128 empirical samples passing sequencing and mapping quality control and 127 retained after the prespecified influence review removed one 40-day outlier.
Experimental unit: Individual tissue sample nested within one selected cohort aquarium for each sampled age group. Cohorts began with 5 to 7 berried females fertilized within eight days of one another, age was assigned from the first female fertilization date and therefore represented a maximum rather than exact age, and the 7-day egg group came from one female. The thesis does not establish independent replicate cohort tanks within an age comparison.
Environment: Commercial red cherry shrimp reared in university cohort aquaria containing equal parts dechlorinated tap and reverse-osmosis water at 22 to 24 C and fed commercial fish wafers ad libitum
Endpoints: global DNA methylation by ELISA, ELISA technical and plate variability, mitochondrial sequence coverage, point heteroplasmy count, haplotype reconstruction, variant-caller and technical-replicate checks.
What it can support: After recalibrating inconsistent ELISA standards, global DNA methylation differed across the four age groups because the 40-day group was higher than juvenile and adult groups; juvenile and adult groups did not differ. The author concluded that this kit-based global measure was not a reliable chronological-age marker. Across approximately 6 kb of mitochondrial sequence in 127 retained shrimp, the number of point heteroplasmies did not differ among seven groups from 7 to 210 days after fertilization once one influential 40-day animal was removed. The second method therefore did not yield a usable age marker under this protocol.
What it cannot support: This was not a hatch-to-death survival or lifespan study. The repeatedly cited approximately 13-month captive lifespan came from another source and was used only to describe sampling coverage. One selected tank per age group leaves age inseparable from cohort-tank, family and shared-history effects; the egg group came from one mother; age windows were based on fertilization rather than observed hatch; sex, family contributions, starting denominators, deaths and censoring were not reported; and only 5 of 9 sampled 210-day animals passed quality control. The methylation result depended on recalibrated controls after only 30 of 100 raw readings fell within the supplied standard curves, with 61.7 percent inter-plate variation before recalibration. Probable amplification of nuclear mitochondrial copies affected roughly 9.5 kb, technical replicate counts agreed only 71 percent of the time and the retained approximately 6 kb represented about 39 percent of the mitogenome. Cross-sectional destructive samples were different animals, not longitudinal records. Do not use this source to claim a 13-month lifespan, label an individual old, infer age from appearance, sell an age test, establish line inheritance or diagnose oxidative damage.
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System observation or method study
Neocaridina identity evidence
Huang et al., 2020. Marine Biotechnology 22: 772-785.
Source taxon as published: Supplier-sourced super red shrimp and chocolate shrimp labelled Neocaridina denticulata, plus a native black-shell strain used in marker development; no voucher or diagnostic identity method was reported
Study shape: RNA from three young super red and three young chocolate shrimp supplied one aggregate sequencing result per phenotype on an Illumina HiSeq 2000. Transcriptome SSR candidates were screened by PCR, sixteen amplified, and five polymorphic loci were reported across offspring from super red, chocolate and reciprocal crosses. Supplementary Figure S4 reports the largest eight females and males from each source strain as parents and three families per cross group.
Experimental unit: The transcriptome comparison appears to contain one pooled library per phenotype rather than independent biological libraries. The paper states 50 offspring in each of four cross groups, but Table 6 frequencies for both reciprocal hybrids occur in eighths and no individual genotype file, missing-call denominator, family allocation or family-aware analysis is supplied.
Environment: Separate supplier strains maintained in a flow-through breeding system at 22 to 25 C, pH 6.5 to 7.0 and GH 3 to 4; tank count, strain allocation and family history before the reported crosses were not defined
Endpoints: assembled transcriptome and annotation, phenotype-group expression counts, SSR discovery and PCR feasibility, allele and genotype counts, heterozygosity and polymorphic information content, genotype frequencies across four cross groups.
What it can support: The study developed five polymorphic transcriptome-derived SSR loci and found different genotype-frequency patterns among its sampled Larmax super red, chocolate and reciprocal-cross groups. The loci are candidates for population and cross validation within the sampled breeding material.
What it cannot support: One apparent pooled RNA library per phenotype cannot support replicated differential-expression inference. The study did not report standardized colour measurements, individual phenotypes, complete genotype denominators, parent-to-offspring assignments, Mendelian segregation, linkage, causal variants, blind classification or an independent validation population. The stated 200-offspring method conflicts with hybrid frequencies in eighths, and the claimed 4.31 percent single-marker and 0.0018 percent combined-marker misrecognition rates are not derived in the article or supplements and were not measured on a held-out test set. Multiple genotype tests were reported without a multiplicity correction and p values were printed as 0.000. These loci do not certify purity, pedigree, ancestry, diversity, Blue Dream, Sunkist, Bloody Mary or any other seller line, and super red is not evidence for equivalence to a current red trade name.
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Controlled experiment
Direct Neocaridina evidence
Li et al., 2020. Journal of Experimental Biology 223: jeb223529.
Source taxon as published: Neocaridina heteropoda
Study shape: MicroRNA, PFK and related metabolism were manipulated with RNA interference, agomirs, antagomirs and hormone treatments.
Experimental unit: Treated animal or prepared biological sample, depending on endpoint
Environment: Laboratory molecular and physiological manipulation
Endpoints: PFK expression, ecdysis interval, body length, metabolites, protein-synthesis signalling.
What it can support: The tested miR-26 and PFK pathway manipulations changed molt interval and body-length outcomes.
What it cannot support: The invasive molecular treatments are not aquarium interventions and do not support a food, mineral or oxygen recipe.
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Controlled experiment
Direct Neocaridina evidence
Tomas et al., 2020. Aquaculture Nutrition 26: 1198-1210.
Source taxon as published: Neocaridina davidi
Study shape: Defined diets were compared for biochemical, pigmentation and male reproductive outcomes.
Experimental unit: Diet treatment culture unit
Environment: Laboratory diet comparison
Endpoints: carotenoids, visible colour, biochemical composition, spermatophore quality.
What it can support: Diet can change measured phenotype and biochemical outcomes under a defined protocol.
What it cannot support: Visible colour change is not inherited grade, genetic purity or proof that one diet is complete for every life stage.
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Controlled experiment
Direct Neocaridina evidence
Tomas, Sganga and López Greco, 2020. Journal of the World Aquaculture Society 51(3): 775-787.
Source taxon as published: Neocaridina davidi
Study shape: Background and shelter conditions were assigned and female pigmentation was followed for 90 days.
Experimental unit: Treatment aquarium
Environment: Laboratory background and shelter treatment
Endpoints: visible pigmentation, astaxanthin.
What it can support: Background can change measured visible pigmentation under a defined protocol.
What it cannot support: It does not establish that dark substrate changes genotype, prevents stress or improves inherited grade.
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Morphology or taxonomy
Neocaridina identity evidence
Han et al., 2019. BMC Genetics 20: 86.
Source taxon as published: Four Neocaridina species or provisional lineages from Taiwan
Study shape: COI sequences, phylogeographic structure and demographic models were used to infer species boundaries and colonization histories.
Experimental unit: Collected specimen, haplotype and sampled locality depending on analysis
Environment: Four hundred seventy-nine specimens from thirty-five Taiwanese localities
Endpoints: COI lineage, haplotype structure, species delimitation, colonization-time inference.
What it can support: The sampled Taiwanese Neocaridina contained multiple geographically structured lineages with different inferred histories.
What it cannot support: Phylogeographic inference does not establish the ancestry, relatedness, purity or species of a commercial colour morph.
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Controlled experiment
Direct Neocaridina evidence
Sganga and López Greco, 2019. Aquaculture Research 50(6): 1564-1573.
Source taxon as published: Neocaridina davidi
Study shape: Maternal colour was measured at defined ages and compared with offspring outcomes.
Experimental unit: Female and associated offspring group
Environment: Laboratory comparison of red-stock females and offspring
Endpoints: female colour, offspring colour, offspring quality.
What it can support: Visible colour changed with age, and maternal colour did not simply predict offspring colour at the measured checkpoint.
What it cannot support: It does not provide a named-morph inheritance chart, breeding value from one parent image or a universal grade age.
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Morphology or taxonomy
Direct Neocaridina evidence
Shih, Cai and Chiu, 2019. ZooKeys 817: 11-23.
Source taxon as published: Neocaridina fonticulata Shih, Cai and Chiu, 2019, described from type and additional material collected at Sheding, Kenting, southern Taiwan
Study shape: The authors described a holotype, multiple paratype lots and additional field material using external morphology and drawings, compared material of Neocaridina ikiensis, measured five eggs from each of four ovigerous females, and analyzed a 658-base-pair mitochondrial COI segment from four Sheding specimens with Bayesian and maximum-likelihood phylogenetic methods and pairwise distances.
Experimental unit: Individual deposited specimen for morphology, four source shrimp for COI that yielded one reported haplotype, and 20 eggs nested within four ovigerous females for egg measurements
Environment: A leaf-litter layer in a small, slow-flowing headwater stream beside a spring outlet on a limestone hill at Sheding, Kenting; the paper reports about 25 C water, pH 7.06 to 7.16 and dissolved oxygen 7.33 to 7.70 mg/L at the collection site, while examined specimens were preserved in 70 to 95 percent ethanol
Endpoints: type locality and deposited material, rostrum and appendage diagnostic characters, male first and second pleopod morphology, egg dimensions, live colour description, COI haplotype and phylogenetic placement, pairwise mitochondrial distance.
What it can support: The combined type material, comparative morphology and sampled COI evidence support recognizing N. fonticulata as a distinct Neocaridina species in the reviewed 2019 treatment. Diagnostic evidence includes the rostrum, pereiopods and male first and second pleopods. The four sequenced shrimp produced one 658-base-pair COI haplotype, LC427866, and the smallest reported K2P distances to sampled N. ketagalan and N. saccam were 5.42 and 5.43 percent.
What it cannot support: The species was then known only from one Sheding locality, so the report does not establish its complete distribution, population size, seasonal habitat range or conservation status; the authors prediction that it may occur in eastern Taiwan was not a sampled result. Several diagnostic characters require an adult male, dissection and specialist comparison, so body colour or one aquarium photograph cannot identify the species or distinguish it from a trade morph. The molecular result used four shrimp, one mitochondrial haplotype and no nuclear locus, cross, genomic ancestry test or blind identification validation. Mitochondrial separation in this sample does not authenticate a seller label, certify a colour line, prove reproductive isolation or identify another animal by appearance. Site temperature, pH and dissolved oxygen are locality observations without temporal replication or an assigned comparison, not aquarium optima, tolerances or care targets. Aquarium-held photographs were for observation after collection, not a husbandry experiment.
Related source material: GenBank COI accession LC427866
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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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System observation or method study
Direct Neocaridina evidence
Sin et al., 2015. General and Comparative Endocrinology 214: 167-176.
Source taxon as published: Neocaridina denticulata linked to the Kenny et al. draft-genome stock; the RNA-source animals were not otherwise identified or described
Study shape: Known arthropod hormone-pathway sequences were searched against the draft genome with TBLASTN and checked by reciprocal BLASTX, characteristic domains and, where identity was uncertain, Bayesian phylogenetic analysis. Selected partial transcripts were amplified from cDNA, cloned and Sanger sequenced. A semiquantitative JHAMT gel used equivalent total-RNA inputs and displayed three individual samples for each reported life-stage or body-region group.
Experimental unit: Gene sequence and draft-genome contig for homology analyses. For the JHAMT expression gel, the caption reports three independent individual shrimp samples per life-stage or body-region group, while the methods describe RNA from several individuals without source, sex, exact age, molt stage or allocation details.
Environment: Computational analysis of the 2014 short-read draft genome plus laboratory RT-PCR and Sanger sequencing from RNA isolated from several animals at different life stages; animal holding conditions were not reported in this article
Endpoints: reciprocal sequence homology, diagnostic protein domains, Bayesian phylogenetic placement, cloned partial cDNA sequence, GenBank accessions KJ200310-KJ200321, KJ579126-KJ579131 and KJ956470-KJ956473, semiquantitative JHAMT RT-PCR band presence across life-stage and body-region samples.
What it can support: The study catalogued candidate orthologues for sesquiterpenoid biosynthesis, binding and degradation; ecdysteroid biosynthesis; and putative hormone regulation or signal transduction in the source draft genome. It validated selected partial transcripts and reported stronger JHAMT gel bands in embryos and juvenile anterior samples than in adult posterior samples under its assay.
What it cannot support: Candidate sequence homology and transcript detection do not demonstrate the encoded protein, enzyme activity, endogenous juvenile hormone, methyl farnesoate or ecdysteroid concentration, tissue-specific function or a causal role in molting or reproduction. Shade was not recovered, which the authors attributed as possibly due to poor recovery of that genomic locus. The JHAMT comparison was a semiquantitative endpoint using equal total-RNA inputs, not qPCR with a reported reference gene, band densitometry or statistical model. Animal source, taxon verification, culture system, sex, exact age, molt and reproductive stage were not reported; the draft genome came from one unvouchered commercial adult. Supplementary alignments, phylogenies and primer details were referenced but were not accessible in this review and were not independently reanalyzed. No live-animal hormone treatment, RNA interference, molt frequency, ecdysis success, growth, survival, fertility or offspring outcome was measured. The paper does not validate a hormone supplement, insect-growth-regulator treatment, mineral dose, breeding intervention, molt diagnosis, commercial-line marker or consumer genetic test.
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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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