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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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
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
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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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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Field observation
Direct Neocaridina evidence
Mitsugi and Suzuki, 2018. Crustacean Research 47: 9-16.
Source taxon as published: Neocaridina davidi
Study shape: Monthly field samples were used for morphology, size-frequency cohorts and inferred life history.
Experimental unit: Monthly river sample
Environment: Tomoe River, eastern Japan, sampled monthly for one year
Endpoints: body and carapace length, sex characters, cohort structure, inferred lifespan.
What it can support: Defined measurement landmarks and field cohort estimates for the sampled population.
What it cannot support: Individuals were not marked from hatch to death, and the inferred lifespan is not an aquarium average or guarantee.
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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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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
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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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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