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
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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Field observation
Direct Neocaridina evidence
Sidorovskyi, 2026. Fisheries Science of Ukraine 1(75): 98-114.
Source taxon as published: Neocaridina davidi
Study shape: Hydrobiological dip-net collections at two coordinates were preserved in 96 percent ethanol and identified by stereomicroscopy; later visits were used to report non-detection after the initial records.
Experimental unit: Field collection event at either of two Kharkiv water bodies; sampling frequency, effort and detection probability were not reported
Environment: Two urban water bodies in Kharkiv, Ukraine, surveyed between June 2020 and December 2025
Endpoints: specimen occurrence, body length, sex count, ovigerous-female count, later non-detection, site context.
What it can support: The article documents 10 and 6 shrimp at one site in 2020 and 2021, seven at a second site in 2024, ovigerous females in every positive collection and later non-detection at both sites.
What it cannot support: Later non-detection does not prove extinction because sampling effort and detectability were not reported. Identification was described as stereomicroscopy without diagnostic character results or a molecular confirmation. The end of warm runoff was not an assigned temperature treatment, and no released source stock was traced. The observations do not establish a thermal threshold, prove release causation or make any climate release-safe.
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Controlled experiment
Direct Neocaridina evidence
Azhar et al., 2025. Planetary Sustainability 3(2): 56-76.
Source taxon as published: Neocaridina davidi, red cherry adults
Study shape: Five approximately 42 percent protein formulas replaced 0, 25, 50, 75 or 100 percent of fish meal with black soldier fly larvae meal. The actual meal contents were 0, 10.8, 21.68, 32.62 and 43.64 percent of the complete dry formulas. Each diet was assigned to five aquaria containing one male and one female, fed to satiation twice daily.
Experimental unit: The 3-litre aquarium, with five aquaria per diet; treatment was assigned by aquarium, but the one-way ANOVA description does not state whether aquarium or animal observations entered each endpoint analysis
Environment: Twenty-five 3-litre aquaria with Java moss during a 60-day feeding trial, with daily water checks, 20 percent water exchange every three days and 50 percent weekly exchange
Endpoints: feed intake, survival, length gain, weight gain, specific growth rate, shed-molt count, ovary measurement, reported offspring count, colour coordinates.
What it can support: The 50 percent fish-meal-replacement formula, which contained 21.68 percent black soldier fly meal, had higher reported length gain, weight gain and specific growth rate than the 75 and 100 percent replacement formulas. The table letters did not distinguish it from the control or 25 percent replacement groups. Survival, ovary measurement, reported offspring count and colour intensity did not differ statistically among diets.
What it cannot support: This was a complete-formula comparison, not a test of adding 50 percent insect meal to finished food. Fish meal, black soldier fly meal, fish oil, corn meal and cellulose changed together, while measured lipid and ash also varied; amino acids, fatty acids and energy were not reported. Every diet contained the same Spirulina and astaxanthin. Feeding was described as both to satiation and predetermined, without an exact ration, uneaten-feed recovery method or leaching correction, and measured water values were not reported. The analysis unit was not explicit. The published molting equation multiplies molts per starting shrimp by 100, while the table reports 4.8 to 7.6 percent without reconciling the scale. Ovary size was calculated as length times width but reported in cm, one figure caption refers to male ovary development, and post-mortality brood denominators are unclear. The study does not establish a universal 50 percent recipe, a whole-insect feeding method, reproductive safety, a colour benefit or a home molting rate.
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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
Li et al., 2025. Turkish Journal of Fisheries and Aquatic Sciences 25(2): TRJFAS25726.
Source taxon as published: Aquaculture-farm Neocaridina davidi; identification method not reported
Study shape: The authors sequenced HIF-1alpha cDNA and profiled mRNA and protein across three gonad stages and seven named embryo stages. They then randomly assigned nominal groups of 300 females and 100 males to intramuscular injection of 600 nL dsHIF-1alpha or dsEGFP. Egg-carrying females observed within three days supplied maternal muscle and removed embryos for qPCR, western blots, glycolytic-gene assays and photographed developmental examples.
Experimental unit: Individual injected adult for assignment, but each treatment occupied a mesh box within one shared-water tank. Embryo expression samples pooled material from three shrimp and reported three repetitions without fully mapping females, broods, embryo counts or pools to each assay. The developmental figure shows one control sequence and three affected examples without the eligible female, clutch or embryo denominator.
Environment: Farm-sourced adults maintained at 27 C with daily compound feed; dsHIF-1alpha and dsEGFP groups occupied mesh boxes within the same shrimp tank, and dissolved oxygen and other water chemistry were not reported
Endpoints: HIF-1alpha cDNA sequence and predicted domains, stage-associated HIF-1alpha mRNA and protein, maternal muscle and embryo HIF-1alpha after dsRNA injection, embryo PFK, HK and PGK transcripts, photographed cleavage progression or arrest.
What it can support: Under the reported assays, HIF-1alpha mRNA and protein were highest at the cleavage stage relative to later named embryo stages. Maternal dsHIF-1alpha injection was associated with lower HIF-1alpha signal in maternal muscle and sampled embryos, lower expression of selected glycolytic transcripts in several cleavage or blastula comparisons, and photographed examples of delayed or arrested early development relative to the dsEGFP sequence.
What it cannot support: The study did not assign oxygen treatments or measure dissolved oxygen, so the HIF name and pathway cannot establish aquarium hypoxia, embryo oxygen demand, aeration need, tolerance or a safe oxygen threshold. Shared tank water prevents independent environmental replication. Taxon identification was not reported, and the methods repeatedly call the shrimp crayfish. The number of egg-carrying females, clutches, embryos, affected embryos, surviving embryos and hatches was not reported, so the developmental frequency and effect size are unknown. Removed-embryo observation conditions and the selection of photographed examples were not fully described. The study did not directly label or trace dsRNA into oocytes or embryos, include an uninjected control, use a second non-overlapping HIF-1alpha dsRNA, report a rescue experiment or measure maternal survival and reproductive output; lower embryo signal therefore does not by itself prove physical intergenerational transfer or exclude off-target and maternal-condition pathways. Sample pooling and biological-replicate mapping are incomplete. Paired t-tests were used for apparently separate treatment groups and repeated stage comparisons without a reported multiplicity correction. Methods define significance at p less than 0.01, text uses p less than 0.05, captions introduce one, two and three-star levels, Figure 6 calls embryo-stage measurements muscle, and Figure 4 is titled protein expression although one panel is mRNA. The seven stage names partly borrow from Macrobrachium literature, and the representative 428-hour control sequence cannot supply a universal hatch countdown or viability test.
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Controlled experiment
Direct Neocaridina evidence
Luan et al., 2025. Biomolecules 15(6): 815.
Source taxon as published: Neocaridina davidi
Study shape: Hepatopancreases from mature red females were pooled in threes, exposed outside the animal to farnesoic acid or methyl farnesoate and compared by RNA sequencing, pathway analysis and qPCR.
Experimental unit: Pooled hepatopancreas sample from three females, with three pooled samples per condition for the 3-hour RNA sequencing comparison
Environment: Excised hepatopancreas tissue cultured at 25 C in Schneider's Drosophila medium with 1 ppm farnesoic acid or methyl farnesoate
Endpoints: differential gene expression, pathway enrichment, qPCR agreement.
What it can support: Excised female hepatopancreas tissue showed different short-term gene-expression responses to the two tested sesquiterpenoid hormones, including digestive-enzyme-associated transcripts after farnesoic acid exposure.
What it cannot support: This was an excised-organ experiment, not a whole-animal feeding, water, growth, molt, reproduction or safety trial. The paper does not document a hormone stock solvent or a matched vehicle control, and its sequencing wording does not make the timing of the control collection fully clear. Pathway labels and gene expression do not establish improved digestion, immunity, growth or vitellogenesis.
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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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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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System observation or method study
Direct Neocaridina evidence
Yan et al., 2024. BMC Genomics 25: 570.
Source taxon as published: Baiyangdian Lake Neocaridina denticulata sinensis; identification method not reported
Study shape: Embryos were assigned by stereomicroscope appearance to four late embryonic stages. Twelve RNA sequencing libraries represented three pooled samples per stage, with about 50 embryos in each sample. Adjacent stages were compared for gene expression and pathway enrichment, and eight selected transcripts were checked by qRT-PCR.
Experimental unit: Pooled RNA sample containing about 50 embryos, with three pools per morphological stage; the number of females or broods contributing to each pool and independence among pools were not reported
Environment: Field-collected adults reared in an indoor recirculating aquaculture system with aeration; temperature, water chemistry, photoperiod, feed and spawning dates were not reported
Endpoints: stereomicroscope stage assignment, whole-embryo RNA sequencing, differential gene expression, GO and KEGG enrichment, putative eye-development, phototransduction, cuticle and molting-related genes, qRT-PCR expression of eight selected transcripts.
What it can support: The study operationally separated late embryos by visible eye development, from thin crescent-shaped black pigment at the first compound-eye stage to oval eyes and later zoea-stage morphology. Whole-embryo expression profiles changed across those stages, including higher late-stage expression of putative visual opsins and enrichment of phototransduction and chitin-related terms.
What it cannot support: The paper did not report embryo ages, elapsed time between stages, time to hatch, incubation temperature, water chemistry, photoperiod, brood count, clutch allocation, egg retention, hatch success, survival, deformity or post-hatch outcome. About 50 embryos were pooled per RNA sample, and unknown family contributions prevent treating every embryo as an independent replicate. Stage and maternal history were confounded. Pathway annotation and expression association do not prove that a gene or hormone caused eye formation, molting or hatching; qRT-PCR of eight selected transcripts was assay confirmation rather than an independent biological replication or functional perturbation. The source labels the final in-egg stage zoea, which must be preserved rather than converted into a hobby life-stage rule. Visible eyes therefore do not provide a universal hatch countdown, prove embryo viability or support a light, temperature, medication or handling recommendation.
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Controlled experiment
Direct Neocaridina evidence
Razekenari et al., 2023. Aquatic Toxicology 259: 106519.
Source taxon as published: Neocaridina davidi
Study shape: Six nominal EE2 concentrations from 0 to 200 micrograms/L were assigned to three aquaria each, with 30 females per aquarium. Ten ready females per treatment were later mated in clean water with unexposed males, eggs were removed for artificial incubation and unexposed offspring were followed for 10 days across five successive spawns.
Experimental unit: The 8-litre exposure aquarium, with three aquaria per concentration; later mating, artificial-incubation and offspring units differed by endpoint
Environment: Eighteen 15-litre laboratory aquaria with 8 litres of useful water at 26 to 28 C, complete water replacement and redosing every two days during 210 days of nominal EE2 exposure
Endpoints: female survival, female weight and length, time to first spawning, interspawn interval, fecundity, egg volume, hatching, hatchling size, 10-day offspring size and survival.
What it can support: Under this nominal exposure protocol, females in the two lowest nonzero treatments were heavier and longer but produced fewer eggs and had lower hatching outcomes, while every nonzero treatment had lower adult survival than the control.
What it cannot support: Exposure concentrations were nominal and not analytically verified. The control was clean water rather than an ethanol-matched vehicle, the written stock-volume calculation conflicts with the reported maximum ethanol percentage and control survival was only 57.3 percent. Full water was replaced every two days, males and offspring were not directly exposed and eggs were artificially incubated. The study does not establish a household threshold, a beneficial low dose, an environmental safe level or an aquarium treatment.
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Controlled experiment
Direct Neocaridina evidence
Zanitti et al., 2023. Archives of Environmental Contamination and Toxicology 85(2): 181-190.
Source taxon as published: Neocaridina davidi
Study shape: Twenty-two aquaria were randomly assigned per treatment. Each began with one ovigerous female and two mature males and received control water or nominal 0.1 or 1 mg/L diclofenac. HPLC-MS/MS checks at 0 and 72 hours in two aquaria per treatment produced overall measured means of 0.0075, 0.1320 and 1.0730 mg/L, respectively. Mortality, molts, egg loss, hatching and ovarian rematuration were checked daily; surviving adults, broods, ovaries and male distal vas deferens supplied later endpoints.
Experimental unit: The 1.5-litre aquarium, with 22 aquaria per assigned treatment
Environment: Sixty-six 1.5-litre laboratory aquaria at 27 C, pH 7.5 and hardness 80 mg/L as CaCO3, with aeration, Java moss, daily ad-libitum feed and complete water replacement plus re-dosing three times weekly during 63 +/- 3 days
Endpoints: mortality, adult specific growth rate, successive spawning timing, hatching, juvenile count and morphology, female ovarian histology, male spermatophore histology.
What it can support: Under this 63-day laboratory protocol, aggregate mortality was 22.7, 39.4 and 45.5 percent, with only the 1 mg/L nominal treatment differing significantly from control. The higher treatment reduced survivor-conditioned female growth, the proportion of females hatching their first exposed brood and the proportion of advanced ovarian oocytes, shortened interspawn timing, and increased abnormal juveniles per female in the small second-spawn sample. Incubation time, male growth, male spermatophore structure and among-treatment hatchling counts did not differ significantly.
What it cannot support: The control was not an analytical zero: its two checks were 0.009 and 0.006 mg/L, for a 0.0075 mg/L mean. The lowest nonzero measured treatment averaged 0.1320 mg/L, and only two nonzero treatments were tested. Analytical checks covered two aquaria per treatment at two times. The first egg cohort had already developed partly before exposure, later brood and tissue endpoints were conditioned on female survival and successful spawning, the second-spawn abnormality result came from few females, and male dependency may not have been fully represented by aquarium in the analysis. Stock came from a dealer-derived laboratory colony without voucher or diagnostic identity confirmation; allocation blinding, feed intake, exposure-period nitrogen chemistry, tissue residues and offspring survival or growth were not reported. The preprint abstract can overstate the juvenile-count result because the paper reports no significant among-treatment decrease. The design does not establish a trace-effect threshold, safe household medication concentration, aquarium treatment, recovery, mechanism or multigeneration outcome. The SSRN and Research Square postings are duplicate preprint versions, not independent replication.
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Controlled experiment
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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Controlled experiment
Direct Neocaridina evidence
Tomas et al., 2021. Acta Zoologica 102(3): 297-309.
Source taxon as published: Neocaridina davidi
Study shape: Reproductive, pigmentation and biochemical outcomes were compared across three assigned temperatures.
Experimental unit: Temperature treatment aquarium
Environment: Laboratory aquaria at 20, 24 and 28 C
Endpoints: survival, ovarian maturation, ovigerous females, weight, colour, spermatophore quality.
What it can support: Lower temperature delayed ovarian maturation in this protocol while survival remained above 90 percent in every treatment.
What it cannot support: It does not make every temperature in the tested range equivalent or establish chronic survival outside the study duration.
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Controlled experiment
Direct Neocaridina evidence
Budi et al., 2020. Turkish Journal of Veterinary and Animal Sciences 44(2): 456-462.
Source taxon as published: Neocaridina davidi
Study shape: Visible ovary stages were paired with histology and fecundity observations.
Experimental unit: Individual female
Environment: Laboratory culture of a red stock at 28 to 29 C
Endpoints: ovarian stage, histology, fecundity.
What it can support: A visible ovary can support female-sex and ovarian-stage observations under the documented criteria.
What it cannot support: A hidden ovary does not prove male sex, and the stage timing is not a universal spawning countdown.
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System observation or method study
Direct Neocaridina evidence
Gomez-Nieves and Gomez-Hernandez, 2020. Revista Investigacion Pecuaria 7(1): 19-23.
Source taxon as published: Laboratory cherry shrimp labelled Neocaridina heteropoda and Neocaridina sp.; identification method not reported
Study shape: Eighteen selected laboratory broodstock, 12 females and six males, were distributed two females to one male across six aquaria and observed for ten months. Length was sampled every 15 days and physicochemical parameters monthly. The paper reports reproductive and size observations but describes only a Kolmogorov-Smirnov normality test.
Experimental unit: Aquarium for the six three-animal groups and individual for some length and reproduction observations; all aquaria received the same management, offspring allocation was not described and animal losses changed density among units
Environment: Six nominal 2-litre glass aquaria near windows at 22 C with Riccia fluitans, fish feed to satiation and a stated dissolved-oxygen target of 2 to 3 mg/L
Endpoints: female and male total length, reported age at sexual maturity, spawns per female, egg count, reported ontogenic duration, temperature and monthly water context.
What it can support: In this small laboratory series, selected adults reproduced at 22 C. The paper reports mean reproductive-phase lengths of 31.3 mm for females and 26.5 mm for males and 2.75 spawns per female during the ten-month observation.
What it cannot support: The source began with selected broodstock averaging 31 mm for females and 26 mm for males, so those animals cannot establish age at first maturity. The methods do not define an F1 starting denominator, hatch dates, offspring allocation or age-assignment procedure even though the abstract and conclusion report maturity in weeks. The abstract gives ranges of 12.1 to 14 weeks for males and 14.2 to 16.4 for females, while the conclusion gives single values of 12.1 and 14.2 weeks. Egg counts of 18 to 33 and ontogenic duration of 18 to 25 days appear in the abstracts but are not supported by a reported observation schedule, denominator or results table. The container description conflicts between 0.20 cubic metres, 0.20 square metres and 2000 mL. Species identification was not reported and a figure labels the animals only Neocaridina sp. Monthly water results were not published. A Kolmogorov-Smirnov normality test does not by itself establish the between-unit differences shown, and the text conflicts between p less than 0.01 and a figure caption using p less than 0.05. Mortality changed density among units. The work cannot validate a maturity age, maturity-size threshold, incubation calendar, universal spawning rate, stocking ratio, 22 C optimum or dissolved-oxygen target.
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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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System observation or method study
Direct Neocaridina evidence
Mahmoud, Sastranegara and Kusmintarsih, 2020. Biodiversitas 21(6): 2396-2402.
Source taxon as published: Market-sourced animals labelled Neocaridina denticulata and Neocaridina palmata; identification method not reported
Study shape: Thirty market-sourced animals per labelled species, 15 male and 15 female, first shared one aquarium per species. Fifteen ovigerous females per species were later described as moving to individual aquaria for egg and hatch observations, after which healthy offspring were reared and measured through a reported maturity sequence. Only descriptive statistics were stated.
Experimental unit: One initial aquarium per labelled species, so species and tank were confounded; the number, size and independence of the later female and offspring aquaria were not reported, and individual offspring tracking was not described
Environment: One 30-litre aquarium per labelled species with powerhead filtration, aeration, white stone, Java moss, twice-daily 40 percent protein pellets, partial water renewal every three weeks and naturally varying 25 to 28 C water
Endpoints: adult and post-hatching total length, egg length and width, attached egg count per female, reported time to life-stage labels, temperature, pH, dissolved oxygen, nitrate, ammonia.
What it can support: Under the reported culture history, the market-sourced groups produced attached eggs and direct-developing shrimp-like young. Fifteen females under each label supplied reported egg-count ranges of 21 to 60 for N. denticulata and 21 to 58 for N. palmata, and photographed post-hatching young measured about 3.3 mm at 24 hours.
What it cannot support: Species identity rested on market sourcing, colour and photographs without a diagnostic key or molecular confirmation. With one initial aquarium per label, species and tank were confounded, preventing an independent species comparison. Female isolation, offspring allocation, starting hatch denominators, survival, losses and repeated-animal tracking were not reported. The stated June 14 to August 10 breeding interval does not reconcile with 75-day and 80-day observations. Tables place ranges in columns labelled Means while also giving separate standard deviations, figures and axes are inconsistently numbered or labelled, and no inferential model is reported. Temperature and pH varied within the same systems rather than being assigned, so plots cannot establish 27 C as an optimum or any pH effect. The water methods, analyte bases and nitrogen interpretation are incomplete, and nitrite named in the abstract is not reported in the tables. Calling the direct-developing young larvae and describing a planktonic phase conflict with the better-resolved N. davidi developmental literature. The source cannot establish a universal 15-day incubation, 75-day maturity calendar, water-quality threshold, species difference or cultivation optimum.
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Controlled experiment
Direct Neocaridina evidence
Sganga and López Greco, 2020. Aquaculture Research 51: 5029-5039.
Source taxon as published: Neocaridina davidi
Study shape: Commercial diets were compared across female reproduction and offspring endpoints.
Experimental unit: Diet treatment group or female, depending on endpoint
Environment: Laboratory commercial-diet comparison
Endpoints: female reproduction, egg outcomes, offspring quality.
What it can support: Diet treatment can change selected reproductive and offspring measures.
What it cannot support: It does not validate every product, a universal ingredient list or a single feeding schedule.
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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
Viau et al., 2020. Aquaculture Research 51: 3847-3864.
Source taxon as published: Neocaridina davidi
Study shape: The full life cycle and juvenile outcomes were followed while comparing conditioned artificial substrates.
Experimental unit: Substrate treatment culture unit
Environment: Designed laboratory biofilm culture across substrate treatments
Endpoints: juvenile survival, growth, biomass, biochemical reserves, reproduction.
What it can support: The tested biofilm community supported the life cycle and substrate treatment changed juvenile outcomes.
What it cannot support: It does not establish that any home-aquarium film is complete food or that tank age measures grazing value.
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Controlled experiment
Direct Neocaridina evidence
Elmas, 2019. Izmir Katip Celebi University MSc thesis.
Source taxon as published: Neocaridina davidi
Study shape: Two commercial grow-out feeds and feeding once or three times daily formed a two-by-two design with three aquaria per treatment, ten shrimp per aquarium and a starting sex ratio of six females to four males.
Experimental unit: The 40-litre aquarium, with three aquaria per feed and frequency combination
Environment: Twelve 40-litre laboratory aquaria at about 26 C across a 12-week feeding trial with twice-weekly 25 percent water changes
Endpoints: feed consumption, weight, total length, carapace length, survival, shed exoskeleton count, egg and hatch measures, carapace lightness and colour coordinates.
What it can support: Under this exact protocol, feeding frequency changed several growth, survival, shed-exoskeleton and reproductive outcomes, while the two tested products also differed for selected reproductive and colour measures.
What it cannot support: The daily ration was not reported, feed consumption differed among groups and the products differed in more than their intended species. Shed exoskeletons were counted by aquarium rather than linked to identified individuals. The thesis does not establish three daily meals for a home colony, isolate an ingredient effect or provide a pellet amount.
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Field observation
Neocaridina identity evidence
Levitt-Barmats et al., 2019. Aquatic Invasions 14(4): 684-702.
Source taxon as published: Neocaridina denticulata, with identity uncertainty discussed by the authors
Study shape: Field occurrence, morphology and mitochondrial markers were combined with an unreplicated laboratory lineage; 18 third-generation larvae were separated for individual molt and growth observations.
Experimental unit: Field specimen or sampling site for field endpoints; individual jar for 18 third-generation growth observations
Environment: Thirty Israeli field locations plus laboratory observation at 20 to 25 C across three generations
Endpoints: field occurrence, morphology, COI and 16S sequences, embryo count, age at first ovigerous stage, molt interval, growth.
What it can support: The study documents a wide field-occurrence range and follows known-hatch third-generation individuals for molt and growth observations over 49 weeks under one laboratory protocol.
What it cannot support: Taxonomic placement within the complex remained uncertain, field occurrence is not a water optimum, and losses plus an incomplete growth series do not provide a replicated hatch-to-death lifespan distribution.
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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
Tomas et al., 2019. Invertebrate Biology 138(1): 17-28.
Source taxon as published: Neocaridina davidi
Study shape: Male reproductive structures and spermatophore formation were described with dissection and microscopy.
Experimental unit: Prepared individual male and tissue specimen
Environment: Laboratory dissection and microscopy
Endpoints: testis and vas deferens anatomy, spermatophore formation, sperm structure.
What it can support: Method-confirmed male reproductive anatomy.
What it cannot support: It does not make internal reproductive organs visible or diagnosable in an ordinary aquarium image.
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Controlled experiment
Direct Neocaridina evidence
Tropea and Lopez Greco, 2019. Canadian Journal of Zoology 97(8): 705-712.
Source taxon as published: Neocaridina davidi
Study shape: Twenty first-spawn broods were randomly assigned by aquarium to an all-male or all-female adult social environment, with 10 aquaria per treatment; one female-environment brood was excluded after unusually high juvenile mortality.
Experimental unit: Brood aquarium, with 10 assigned aquaria per treatment before one exclusion
Environment: Twenty small laboratory aquaria at 27 C, each containing one newly hatched brood with seven adult males or seven adult females for 50 days
Endpoints: juvenile phenotypic sex ratio, 50-day survival, body mass, newly hatched brood count.
What it can support: Seven adult males versus seven adult females did not shift the juvenile phenotypic sex ratio away from 1:1 or produce a treatment difference in sex ratio under this 50-day protocol.
What it cannot support: The study did not compare a no-adult control, different adult densities, temperature effects, molecular sex markers or reproductive success after day 50. A null result in these two social treatments does not prove one universal genetic sex-determination mechanism.
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Controlled experiment
Direct Neocaridina evidence
Baliña et al., 2018. The Biological Bulletin 234(3): 139-151.
Source taxon as published: Neocaridina davidi
Study shape: Juveniles were assigned to 28 or 33 C, and part of the warm group was later transferred to 28 C.
Experimental unit: Temperature treatment aquarium
Environment: Laboratory culture at 28 or 33 C for up to 200 days
Endpoints: ovarian maturation, spawning, recovery after transfer.
What it can support: The tested 33 C treatment suppressed ovarian maturation and spawning, and transferred females later resumed reproductive development.
What it cannot support: This was not an acute lethal test and does not define a safe interval, home heater setting or lifetime boundary.
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Field observation
Direct Neocaridina evidence
Jablonska et al., 2018. Knowledge and Management of Aquatic Ecosystems 419: 14.
Source taxon as published: Neocaridina davidi identified by morphology and a 610-base-pair COI fragment
Study shape: Qualitative hydrobiological hand-net samples collected field shrimp in July 2003, November 2013 and June 2017. Morphology was compared with published descriptions. COI sequences from field and amateur-aquarium animals were aligned with five Taiwan and Hawaii reference sequences, assigned to haplotypes and placed in a neighbour-joining tree.
Experimental unit: Collected animal nested within three field collection events and one aquarium comparison source. Only June 2017 reports standardized effort: one square metre sampled for approximately 15 minutes. The 2003 formaldehyde-preserved sample was unavailable for COI analysis.
Environment: A roughly 4.5 km artificial canal connected to the lower River Oder near Gryfino, Poland, receiving heated Dolna Odra Power Station cooling water, with fine sand and mud substrate, partial concrete banks and submerged vegetation
Endpoints: field occurrence by year, sex and ovigerous status, body length and diagnostic morphology, 610-base-pair COI sequence, haplotype assignment, neighbour-joining placement.
What it can support: Fifteen field shrimp were found in three positive collection years spanning 2003 to 2017, including one ovigerous female in November 2013. Morphology and seven field COI accessions supported N. davidi identity. The seven field and six amateur-aquarium sequences generated by the study shared one COI haplotype, supporting close mitochondrial similarity within that comparison.
What it cannot support: The three positive years were not continuous monitoring, and qualitative sampling with mostly unreported effort cannot estimate abundance, density, occupancy, population trend or detection probability. One ovigerous female supports reproductive condition, not observed hatching, recruitment or proof that local reproduction maintained the records. The authors explicitly retained repeated yearly introductions as an alternative. The canal was artificially heated; the paper reports 27 to 29 C in June-July and 15 C in November but no winter minimum, winter sample, thermal time series or comparison site. It cannot establish cold tolerance, winter survival, a home temperature range or a 27 to 28 C breeding optimum. A shared short mitochondrial haplotype does not trace an aquarium source, seller, release event, nuclear ancestry or transfer direction. The study measured no survival, growth, water chemistry beyond temperature, ecological impact, pathogen transfer or control outcome. Failure to find a threat is not proof of no impact, and field occurrence never makes release safe.
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Controlled experiment
Direct Neocaridina evidence
Namaei Kohal et al., 2018. Journal of Applied Phycology 30: 431-443.
Source taxon as published: Neocaridina davidi
Study shape: Six formulated diets containing 0, 1, 3, 5, 8 or 10 percent Arthrospira meal were assigned to three aquaria each. Diets were approximately equal in protein and energy because Arthrospira replaced part of the fish meal; shrimp received 3 percent of wet biomass daily in three feedings and females were followed through eight spawning events.
Experimental unit: Aquarium, with three aquaria per diet; the independent incubation unit for detached eggs is not clearly described
Environment: Eighteen aerated aquaria at 26 to 28 C and pH 7.0 to 7.2 across an 11-month feeding trial, with complete water replacement every 10 days
Endpoints: weight and length growth, feed conversion ratio, survival, whole-body composition, time to maturity, inter-spawning interval, egg production, relative and actual fecundity, hatching percentage, hatchling length.
What it can support: Under this exact formulated-diet protocol, several growth, female-survival and reproductive measures were higher at 8 or 10 percent Arthrospira meal than in the control, while the reported relative-fecundity advantage declined after the sixth spawning.
What it cannot support: The percentages describe dry-diet formulation, not an aquarium dose or addition to a finished food. Arthrospira replaced fish meal, control female survival was only 25.7 percent after 11 months, eggs were detached and artificially incubated, and incubation replication is unclear. The study does not establish a universal 8 to 10 percent recipe or validate a current commercial product.
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Field observation
Direct Neocaridina evidence
Schoolmann and Arndt, 2018. Limnologica 71: 1-7.
Source taxon as published: Neocaridina davidi
Study shape: Repeated field sampling described population structure, reproduction and dispersal.
Experimental unit: Field samples from one invaded stream system
Environment: Thermally affected German stream followed for 27 months
Endpoints: size-frequency structure, ovigerous females, population persistence, dispersal.
What it can support: A reproducing feral population persisted and dispersed under the recorded field conditions.
What it cannot support: Field estimates do not establish aquarium lifespan, a species temperature optimum or a release-safe climate.
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Controlled experiment
Direct Neocaridina evidence
Tropea, Lavarías and López Greco, 2018. Zoology 130: 57-66.
Source taxon as published: Neocaridina davidi
Study shape: Male or female presence and access to physical, visual or short-range cues were manipulated while ovarian development was followed.
Experimental unit: Focal juvenile female within an assigned social or cue-access treatment
Environment: Laboratory social and cue-access treatments involving juvenile focal females
Endpoints: ovarian growth, time course of maturation, effect of conspecific sex and contact.
What it can support: Male presence accelerated ovarian growth and physical contact was required for the reported effect, while female presence delayed ovarian growth.
What it cannot support: It does not establish a home colony sex ratio, prove a distant pheromone effect or let rapid swimming diagnose mating readiness.
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Controlled experiment
Direct Neocaridina evidence
Pantaleão et al., 2017. Aquaculture Research 48(2): 553-569.
Source taxon as published: Neocaridina davidi
Study shape: Animals were followed through twelve post-hatching stages with microscopy and defined carapace measurements.
Experimental unit: Individual developmental observations within one culture protocol
Environment: Laboratory aquarium culture at 25 C
Endpoints: appendage development, sex differentiation, carapace length, first attached eggs.
What it can support: Stage-specific external development, the method used to distinguish sex and first reproduction observed under the tested protocol.
What it cannot support: It does not promise that home shrimp can be sexed at a fixed age, reach a universal breeding size or carry eggs by a fixed day.
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Controlled experiment
Direct Neocaridina evidence
Serezli et al., 2017. Fresenius Environmental Bulletin 26(12): 7575-7579.
Source taxon as published: Neocaridina davidi
Study shape: Three aquaria per temperature were maintained for 180 days and the paper reports sex classification of 30 offspring from each temperature group.
Experimental unit: Aquarium for the temperature treatment, with three aquaria per temperature
Environment: Nine laboratory aquaria assigned across 20, 23 and 26 C for 180 days
Endpoints: reported offspring sex ratio, survival.
What it can support: The study reported different offspring sex proportions among its three temperature groups.
What it cannot support: Limited reporting, a small tank design and conflict with another direct study make this a replication target, not a sex-production recipe.
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Controlled experiment
Direct Neocaridina evidence
Vazquez et al., 2017. Journal of Crustacean Biology 37(4): 367-373.
Source taxon as published: Neocaridina davidi
Study shape: Newly hatched shrimp were randomly assigned to 2.5, 5 or 10 animals/L for 90 days in 2 L vessels containing 5, 10 or 20 shrimp. The results table reports 8, 9 and 7 vessel replicates, respectively. A second experiment randomly assigned ten 30-day juveniles to each of five mixed-sex, five female-only and five male-only vessels at 5 animals/L.
Experimental unit: Two-litre vessel. Shrimp in each vessel were weighed together every 30 days in the density experiment, so the repeated response was vessel mean mass rather than an individually tracked trajectory. Whole-vessel biochemical homogenates were pooled across replicates when biomass was insufficient.
Environment: Two-litre culture vessels at 27 +/- 1 C with 0.7 g Java moss, continuous aeration and weekly complete water replacement
Endpoints: vessel mean wet mass through 90 days, model-estimated survival, phenotypic sex ratio, presence of ovigerous females, whole-vessel glycogen, lipid and protein concentration, mixed-sex versus monosex vessel mean mass and survival.
What it can support: Under the tested juvenile grow-out conditions, vessel mean mass diverged after day 30 and was lower at 10 animals/L than at 2.5 or 5 animals/L by days 60 and 90. Final females at 2.5 animals/L were 45 percent heavier than females at 10 animals/L, males at 2.5 animals/L were heavier than males at both higher densities, and model-estimated 90-day survival did not differ statistically among densities. Monosex versus mixed culture did not change mean mass or survival in the second experiment.
What it cannot support: This does not establish shrimp per gallon, a home-aquarium carrying capacity, a disease or welfare threshold, or a minimum starter group. The units were small juvenile grow-out vessels with daily food described as ad libitum and approximately 4 percent of vessel biomass, continuous aeration, weekly complete water replacement and removal of every new offspring. Individuals were not tracked, starting vessel counts were unequal across densities, brood and family contributions were not reported, food intake was not measured, and ovigerous presence was recorded without egg, hatch or recruitment outcomes. Survival percentages were model estimates rather than raw tank-wide guarantees. Biochemical samples sometimes pooled vessel replicates, weakening the original unit boundary. Ninety days does not represent a mixed-age, multigeneration colony or lifetime health.
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Morphology or taxonomy
Direct Neocaridina evidence
Sganga, Piana and López Greco, 2016. Zootaxa 4196(1): 120-128.
Source taxon as published: Neocaridina davidi
Study shape: Carapace shape was analysed in 92 animals and second abdominal pleura in 61 after sex assignment from pleopod morphology.
Experimental unit: Individual measured shrimp
Environment: Laboratory geometric morphometric study of cultured animals
Endpoints: carapace size and shape, rostrum shape, second pleuron dimensions, sex-associated overlap.
What it can support: Adult females were larger on average and differed in several shapes, while male carapace shape overlapped juvenile female shape.
What it cannot support: Average adult differences do not confirm the sex of every individual, especially a juvenile or an animal shown from one angle.
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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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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
Direct Neocaridina evidence
Tropea and López Greco, 2015. The Biological Bulletin 229(3): 243-254.
Source taxon as published: Neocaridina davidi
Study shape: Female condition, eggs and offspring were measured across five or six consecutive spawns and compared with non-reproducing females where relevant.
Experimental unit: Individual female or spawn, with offspring followed under the study protocol
Environment: Laboratory culture following females, eggs and offspring across repeated spawnings
Endpoints: eggs per spawn, egg biochemical composition, hatchlings, hatchling size, juvenile growth, female energy and mass.
What it can support: Later spawns had lower egg lipid and energy measures, and reproduction was associated with lower female energy and body mass under the protocol.
What it cannot support: It does not show that every later brood is inferior, diagnose poor care from female size or create a universal brood limit.
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Controlled experiment
Direct Neocaridina evidence
Tropea, Stumpf and López Greco, 2015. PLOS ONE 10(3): e0119468.
Source taxon as published: Neocaridina heteropoda heteropoda
Study shape: Egg incubation and 90-day juvenile culture were compared across three assigned temperatures.
Experimental unit: Brood or aquarium treatment unit, depending on endpoint
Environment: Laboratory aquaria at 24, 28 and 32 C
Endpoints: incubation duration, hatchlings per female, growth, survival, ovigerous females.
What it can support: Temperature changed incubation time and the shape of early growth and reproduction under the tested conditions.
What it cannot support: It did not establish a universal ideal temperature, a lifetime effect, a lethal limit or a rule that warmer always means better growth.
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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
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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Field observation
Direct Neocaridina evidence
Jeong, Oh and Ma, 2006. Korean Journal of Fisheries and Aquatic Sciences 39(Special Issue): 198-202.
Source taxon as published: Wild Neocaridina denticulata denticulata collected around Wolchulsan in Yeongam County, Korea
Study shape: Only ovigerous field females were selected for brood analysis. Carapace length was measured to 0.01 mm, every attached egg was counted and classified as non-eyed or eyed, five eggs per female were sampled for long- and short-axis volume estimates, and females plus eggs were dried for 48 hours at 80 C for reproductive-output calculations. Separate monthly groups of 48 to 60 females supplied gonadosomatic-index measurements.
Experimental unit: The individual selected ovigerous female for brood, egg-volume and reproductive-output results; the monthly female sample for the cross-sectional gonadosomatic series. Table 1 reports 43 Neocaridina females, while Table 2 reports 35 non-eyed-stage and 48 eyed-stage females.
Environment: One Korean valley-stream population collected by scoop net during 2001, fixed in 5 percent neutral formalin and analyzed in the laboratory
Endpoints: carapace length, attached egg count, egg volume, female and egg dry weight, reproductive output, monthly gonadosomatic index, ovarian dry weight by embryo stage.
What it can support: Among 43 selected ovigerous females measuring 5.69 to 7.85 mm carapace length, broods ranged from 47 to 117 eggs with a reported mean of 82 +/- 18. Mean sampled egg volume was 0.70 mm3, and reported reproductive output averaged 41.84 percent. Monthly gonadosomatic index was 3.8 to 4.3 from January through March, peaked at 27.5 in May, remained above 10 through July and declined from August. At a common carapace-length slope, ovarian dry weight was higher in females carrying eyed rather than non-eyed eggs, supporting ovarian rematuration during embryo development in this field sample.
What it cannot support: The published taxon is N. denticulata denticulata from one wild Korean population, not captive N. davidi or a selected aquarium line. The article does not report a taxonomic key or voucher, exact collection dates, effort, water temperature, chemistry, food, female source denominator or annual replication. Selection of ovigerous females prevents estimating prevalence, mating success or population fecundity. Preservation and destructive cross-sectional sampling mean individual females were not followed through hatch or a second brood; ovarian rematuration supports only the potential for consecutive spawning. Egg fertilization, retention, hatch, juvenile survival and brood loss were not measured. The monthly field pattern is not a temperature treatment, aquarium breeding calendar, clutch forecast or care target.
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Morphology or taxonomy
Direct Neocaridina evidence
Naruse, Shokita and Cai, 2006. Proceedings of the Biological Society of Washington 119(1): 25-31.
Source taxon as published: Neocaridina iriomotensis Naruse, Shokita and Cai, 2006, described from the upper reaches of Nishifunatsuki in the Nakama River, Iriomote Island, Japan
Study shape: The listed material contains one male holotype, 41 paratypes and 49 non-type specimens collected on seven dates from November 1999 through June 2000. Only specimens above 4 mm carapace length were used for the description. Characters were drawn with a camera lucida and measured to 0.01 mm; proportional summaries report medians, ranges and varying sample sizes from 73 to 83. The treatment compared the new species with published N. anhuiensis and topotypic N. ishigakiensis morphology.
Experimental unit: Individual field specimen for morphological ratios and qualitative seasonal comparison; the paper does not report the female or egg denominator underlying its egg-size and 21-to-82 clutch ranges
Environment: Field-collected from the lower part of a headwater reach in the upper Nakama River, described as about 5 to 10 m wide, approximately 1 m deep and slow-flowing, where shrimp occurred on aquatic vegetation and tree roots with Macrobrachium shokitai; no water chemistry or temperature was reported
Endpoints: deposited type and non-type material, rostrum armature and length, pterygostomian spine, pereiopod proportions, season-associated third-pereiopod form, male first and second pleopod morphology, egg dimensions and reported clutch range, habitat and distribution description.
What it can support: The 2006 morphological treatment supports recognizing N. iriomotensis within its stated species concept and distinguishes its sampled material from N. anhuiensis and N. ishigakiensis using combinations of rostrum, pterygostomian-spine, pereiopod and male pleopod characters. It also documents that the third pereiopod showed sex-associated form in summer specimens but no sexual difference in winter specimens, demonstrating that season can affect the availability of an identification character.
What it cannot support: All listed material came from one named upper-reach locality in the Nakama River, and qualitative wording that the shrimp was common lacks sampling effort and a population denominator. No molecular analysis was included; proposed headwater ecomorph populations had intermediate rostrum lengths and were retained only tentatively pending molecular and ecological study. Published comparisons relied partly on earlier descriptions rather than a balanced multi-population validation, character sample sizes varied because not every specimen contributed every ratio, and no blind key-performance test was conducted. Egg and clutch ranges lack female, clutch and egg denominators. The paper reports no water temperature, pH, hardness, conductivity, dissolved oxygen, nitrogen measurements, diet, culture comparison or aquarium outcome. It cannot identify an aquarium animal from colour or one photograph, authenticate a seller line, prove genetic separation or reproductive isolation, map the complete species range, establish population abundance, or supply a care target, breeding forecast or seasonal aquarium rule.
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Field observation
Direct Neocaridina evidence
Oh et al., 2003. Crustaceana 76(8): 993-1015.
Source taxon as published: Neocaridina denticulata denticulata from the Young-am stream in Korea
Study shape: Five replicate 0.4-square-metre littoral vegetation samples were collected monthly with a 0.5 mm scoop net; size frequencies, gonads, broods and eggs were used for seasonal population and reproductive estimates.
Experimental unit: The 0.4-square-metre box sample, with five scheduled samples per month
Environment: A permanent temperate stream sampled monthly from July 1999 through June 2001
Endpoints: field density, carapace length frequency, sex ratio, ovigerous season, brood count, egg volume and mass, ovarian stage, modeled growth and maturity.
What it can support: This population showed seasonal reproduction from May through September, measured broods of 53 to 146 eggs and different maturity estimates when mature ovaries and ovigerous status were used as endpoints.
What it cannot support: This is a field population of the published N. denticulata subspecies, not an N. davidi aquarium trial. Growth and cohort parameters were modeled from monthly length frequencies, animals were not tracked individually and field density is not aquarium capacity.
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Field observation
Direct Neocaridina evidence
Niwa and Hamano, 1990. Researches on Crustacea 19: 43-54.
Source taxon as published: Neocaridina denticulata from the Sugow River in Japan
Study shape: Daytime sweep-net collections lasting two to three hours were measured to 0.1 mm body length and grouped into inferred seasonal cohorts; a later three-mesh comparison examined size selectivity.
Experimental unit: Collection date and sampled river reach, with individual measurements nested within each collection
Environment: An approximately 50-square-metre vegetated river reach sampled 26 times from May 1985 through May 1986
Endpoints: body-length frequency, sex ratio, ovigerous season, smallest ovigerous length, inferred cohort timing and growth.
What it can support: Across 19,084 collected shrimp, newly hatched animals appeared in samples from June through August and length-frequency analysis supported three seasonal cohort types in this river population.
What it cannot support: Cohorts were inferred rather than composed of marked animals, daytime effort was time based, net selectivity affected small animals and sex below 8 mm was allocated using an estimated ratio. The results do not establish N. davidi aquarium age, maturity or lifespan.
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