Controlled experiment
Direct Neocaridina evidence
Antcliffe et al., 2026. Royal Society Open Science 13(3): 251712.
Source taxon as published: Neocaridina davidi, with Palaemon varians as a marine comparison
Study shape: Oxygen saturation was followed for mostly 48 hours in open and closed carcass containers with carcass-free controls. A separate group of freshwater carcasses was scored daily for ten days for internal and external post-mortem change.
Experimental unit: Individual adult carcass in one container: three open and five closed freshwater oxygen containers; controls were carcass-free containers but their per-condition allocation was not clearly reported. A separate 12-carcass freshwater sequence supplied daily morphology scores.
Environment: Individual euthanized adult carcasses in 20 mL closed or 100 mL open reverse-osmosis deionized freshwater at 21 C, plus a separate open-water anatomical decay sequence
Endpoints: oxygen saturation near or around the carcass, hourly oxygen change, post-mortem internal and external morphology scores.
What it can support: In the tested 20 mL closed freshwater vials, a single adult N. davidi carcass drove oxygen saturation to complete drawdown within about one day. Open 100 mL freshwater bottles remained above full anoxia but every carcass run became at least dysoxic by 48 hours. Separate daily scoring showed rapid internal tissue opacity before major external collapse.
What it cannot support: The one-carcass-per-20-or-100-mL reverse-osmosis-water systems had no filter, substrate, plants, aeration, scavengers or living tankmates and cannot be scaled to a home aquarium, a tank-wide oxygen prediction or a removal deadline. Oxygen was reported as percent saturation and measured locally or in the tiny vial, not as aquarium mg/L. Freshwater oxygen groups were small, with three open and five closed containers, control allocation was unclear, oxygen and anatomy sequences used different euthanasia methods, and the work did not measure disease transmission, tankmate outcomes or cause of death. Clove oil and magnesium chloride methods are not home euthanasia protocols.
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Controlled experiment
Neocaridina identity evidence
Feng et al., 2026. PLOS ONE 21(2): e0342746.
Source taxon as published: Neocaridina denticulata sinensis
Study shape: The NdTryp gene was cloned, expression was profiled across tissues and nine embryonic stages, in situ hybridization localized transcripts, RNA interference was followed by injected Vibrio parahaemolyticus challenge, and recombinant protein activity was tested in vitro.
Experimental unit: Individual shrimp for animal endpoints, usually three animals per group or time point; the wording for pooled embryonic samples does not clearly separate the three individuals from the stated three biological replicates, and recombinant-enzyme measurements used technical triplicates
Environment: Market-sourced shrimp acclimated for seven days in recirculating laboratory aquaria at 25 C, followed by molecular assays, injected bacterial challenge and isolated recombinant-protein tests
Endpoints: tissue and developmental expression, hepatopancreatic localization, RNA interference knockdown, histopathology after bacterial injection, recombinant enzyme activity.
What it can support: NdTryp expression was highest in the hepatopancreas and localized mainly to R cells and epithelial cells lining its tubules. Expression appeared late in embryonic development, rose after the tested bacterial injection, and knockdown before challenge was associated with more severe hepatopancreatic damage than the dsEGFP challenged control.
What it cannot support: The injected laboratory challenge, small animal groups and ambiguous embryonic pooling do not establish natural infection, diagnosis, treatment, survival or pathogen clearance. The recombinant enzyme optima and 20 mM ion assays are not aquarium pH, temperature, mineral, copper or cadmium targets. No feeding trial tested trypsin as an additive, digestibility aid or growth treatment, and no explicitly described uninfected RNA interference histology control isolates knockdown injury from infection interaction.
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Controlled experiment
Direct Neocaridina evidence
Gorol et al., 2026. The European Zoological Journal 93(1): 563-582.
Source taxon as published: Neocaridina davidi
Study shape: As a continuation of the 2025 experiment, adults received clean water or nominal 1, 5 or 10 mg/L BPA for 24, 48 or 72 hours. Qualitative methods used organs from five animals per group and quantitative methods used five cell suspensions per group.
Experimental unit: Individually exposed adult and its derived organ or cell preparation; this continued the earlier exposure program and is not evidence of an independent replication
Environment: Adult shrimp held individually in 0.5-litre containers at 21 C, pH 7 and GH 10, fed ad libitum, with excrement removed every 12 hours and fresh nominal exposure water prepared for each animal
Endpoints: reactive oxygen species, apoptosis and necrosis, caspase and Bcl-2 states, DNA damage, mitochondrial structure and membrane potential, reduced glutathione.
What it can support: The assigned acute BPA treatments changed multiple intestine and hepatopancreas cell endpoints. Reactive-oxygen signals were highest after 24 hours and then declined, while cell-death, mitochondrial and DNA-damage responses varied by organ, concentration and time; hepatopancreas DNA damage was higher after 48 hours at every tested concentration.
What it cannot support: This is a continuation of the same nominal concentration and exposure design as the 2025 publication, not independent confirmation. Concentrations were not analytically verified, controls were clean-water organ groups rather than an explicitly time-matched series, and five method preparations per group do not establish the total number of independent animals across assays. Declining reactive-oxygen signal does not prove recovery because other cellular endpoints changed on different timelines. No whole-animal survival, behaviour, reproduction, chronic effect or recovery endpoint was measured. It does not establish an aquarium BPA threshold, a general plastic-safety rule or a diagnosis from visible symptoms.
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Controlled experiment
Direct Neocaridina evidence
Guz et al., 2026. Journal of Fish Diseases 49(7): e70127.
Source taxon as published: Ornamental Neocaridina spp.
Study shape: Five isolates were identified biochemically and by mass spectrometry, screened for virulence genes, challenged back into shrimp and tested for drug susceptibility.
Experimental unit: Infection challenge group or bacterial isolate, depending on endpoint
Environment: Diagnostic isolation followed by laboratory pathogenicity challenges and antimicrobial susceptibility testing
Endpoints: isolate identity, virulence genes, challenge mortality, lesion reproduction, antimicrobial susceptibility.
What it can support: The tested isolates reproduced disease in challenge animals with isolate-specific mortality and multidrug-resistance patterns.
What it cannot support: A dark lesion or lethargy cannot identify Aeromonas, and isolate resistance results do not authorize an aquarium antibiotic or dose.
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Review or synthesis
Broader ornamental-shrimp review
Holroyd et al., 2026. Conservation Physiology 14(1): coag046.
Source taxon as published: Tropical ornamental shrimp, including Neocaridina literature
Study shape: Published physiology, trade and transport evidence was synthesized and major gaps were identified.
Experimental unit: Published source, not a new animal experiment
Environment: Review of aquarium trade and transport evidence
Endpoints: temperature evidence, oxygen evidence, carbonate chemistry, nitrogen waste, health assessment.
What it can support: The review maps major species-specific evidence gaps and relevant transport stressors.
What it cannot support: It does not validate a drip rate, package, forecast gate, water target or treatment for Neocaridina.
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Controlled experiment
Direct Neocaridina evidence
McIvor et al., 2026. Aquaculture Research 2026(1): 6618042.
Source taxon as published: Neocaridina davidi, red cherry variant
Study shape: Three aquaria per treatment received 0, 20, 2000 or 20000 smooth 38 to 45 micrometre particles per litre, starting with 20 females and five males per aquarium.
Experimental unit: Thirty-litre aquarium, with three aquaria per concentration
Environment: Twelve 30-litre aquaria with sponge filtration and five months of smooth polystyrene-sphere exposure
Endpoints: particle ingestion, monthly mortality, counts by size class, post-exposure mortality.
What it can support: The two higher tested concentrations increased mortality and changed population size-class counts under the five-month exposure.
What it cannot support: Fecundity, gonads and individual growth trajectories were not measured, so size-class counts do not directly establish slower reproduction or individual growth rates; the treatments are not aquarium safety thresholds.
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Controlled experiment
Direct Neocaridina evidence
Chen et al., 2025. Antioxidants 14(6): 635.
Source taxon as published: Neocaridina denticulata
Study shape: One hundred twenty juveniles were assigned to a basal diet or feed containing 0.4, 4 or 40 micrograms methyl farnesoate per kilogram, with repeated length measurements and end-point biochemical and transcriptomic assays.
Experimental unit: Not recoverable as an independent tank unit: the paper describes four diet groups but does not report replicate tanks; RNA sequencing used three individual shrimp per group
Environment: Forty-day laboratory feeding study at 26 C with a 14-hour light and 10-hour dark cycle and recirculating filtration
Endpoints: body length growth rate, digestive enzyme activity, oxidative-stress assays, whole-animal gene expression.
What it can support: The reported length, enzyme, oxidative-stress and gene-expression outcomes varied with dietary methyl farnesoate concentration and time in the study population.
What it cannot support: The paper does not describe replicated tanks, so diet may be confounded with one group or culture system and individual shrimp do not repair that treatment-level replication problem. The control feed is not explicitly described as ethanol-vehicle matched, and prepared-feed hormone concentration, stability, leaching, intake and internal dose were not verified. Survival, reproduction, long-term safety and withdrawal were not established. This study cannot support a home additive, water dose, growth product or breeding treatment.
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Controlled experiment
Neocaridina identity evidence
Feng et al., 2025. Comparative Immunology Reports 8: 200226.
Source taxon as published: Market-source Neocaridina denticulata sinensis without reported voucher or diagnostic identity confirmation
Study shape: The NdCrus1 cDNA was cloned, transcript abundance was compared across ten tissues from three healthy shrimp, recombinant NdCrus1 was expressed in Escherichia coli, and the purified protein was tested against bacterial growth, selected proteases, chitin materials, reducing conditions and heat pretreatment.
Experimental unit: Individual shrimp or tissue RNA for expression work and a bacterial culture, protein reaction or material-binding preparation for in vitro endpoints; three replicates were commonly reported, but biological and technical replication was not mapped clearly for every assay
Environment: Market-source shrimp held at 25 C in aerated circulating water, followed by tissue expression work and isolated recombinant-protein assays
Endpoints: tissue transcript abundance, bacterial growth and inhibition zones, purified and bacterial protease inhibition, chitin and chitosan binding, activity after reducing-agent exposure, activity after purified-protein heat pretreatment.
What it can support: NdCrus1 was cloned and its recombinant protein inhibited proteinase K and proteases in selected bacterial preparations, showed qualitative binding to chitin materials, and lost inhibitory activity after stronger reducing or heat pretreatments. It did not significantly inhibit growth of Bacillus subtilis or Vibrio parahaemolyticus in the reported bacterial-growth assay.
What it cannot support: The study did not challenge living shrimp, manipulate NdCrus1 in living shrimp or measure survival, clinical signs, pathogen load, clearance, transmission or protection. An antimicrobial-peptide family label and in vitro protease inhibition do not establish disease resistance, diagnosis or treatment. Qualitative chitin and chitosan binding does not validate a feed, prebiotic, supplement, molting aid or dose. The 30 to 100 C pretreatments were applied to purified recombinant protein and are not shrimp temperature-tolerance or aquarium-care tests. Tissue expression used only three animals and the ovary and testis sampling allocation was unclear. No voucher, morphological diagnosis or molecular identity confirmation was reported for market-source animals.
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Controlled experiment
Direct Neocaridina evidence
Gorol et al., 2025. The European Zoological Journal 92(1): 390-409.
Source taxon as published: Neocaridina davidi
Study shape: Adults received clean water or nominal 1, 5 or 10 mg/L BPA for 24, 48 or 72 hours. One animal occupied each container, and five specimens from each group were selected for each microscopy, histochemistry, fluorescence or cell-cycle method.
Experimental unit: Individually exposed adult in a 0.5-litre container; method-specific samples used five animals per group, while reuse or independence across methods was not stated clearly
Environment: Adult shrimp held individually in 0.5-litre plastic containers at 21 C, pH 7 and GH 10, fed ad libitum, with excrement removed every 12 hours and fresh nominal exposure water prepared for each animal
Endpoints: intestine and hepatopancreas ultrastructure, autophagic and acidic structures, lipid, protein and polysaccharide reserves, regenerative-cell ultrastructure, Ki67-positive cells, cell-cycle phase proportions.
What it can support: The assigned acute BPA treatments produced organ-, time- and concentration-associated changes in midgut cell structure, autophagic signals, reserve material, regenerative-cell measures and cell-cycle distributions. The highest treatment produced extensive intestinal degeneration after 72 hours.
What it cannot support: Concentrations were nominal and not analytically verified. The clean-water control came from continuous culture rather than an explicitly time-matched control series, and the number of distinct animals across the many methods was not clear. The study lasted 24 to 72 hours, used adult tissue and cell endpoints, and did not report whole-animal survival, behaviour, reproduction, recovery or a chronic exposure response. It does not establish a household BPA threshold, diagnose damage from an aquarium observation or show that an unnamed plastic product caused exposure.
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Controlled experiment
Direct Neocaridina evidence
Hugger, 2025. Master thesis, University of Vienna.
Source taxon as published: Neocaridina davidi
Study shape: Ionic silver, PVP-coated nanospheres and uncoated nanopowder were compared with measured dissolved silver and animal accumulation.
Experimental unit: Mesocosm exposure unit, not each shrimp sharing the exposure
Environment: Mesocosm exposures in artificial Danube water
Endpoints: 24-hour mortality, median lethal estimate, dissolved silver, whole-animal accumulation.
What it can support: The three tested silver forms had different dissolution and acute mortality relationships under the mesocosm chemistry.
What it cannot support: A master thesis acute result is not a chronic safe value, copper proxy, product limit or household aquarium target.
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Morphology or taxonomy
Direct Neocaridina evidence
Kusmintarsih et al., 2025. E3S Web of Conferences 609: 02003.
Source taxon as published: Market-labeled Neocaridina denticulata and Neocaridina palmata; no host-identification method reported
Study shape: One hundred market-labeled N. denticulata and 100 market-labeled N. palmata were transported in market aquarium water. Gill and cephalothorax material was scraped onto slides and examined by microscopy for organisms reported as sessile bdelloid rotifers, Vorticella sp. and Zoothamnium sp.
Experimental unit: Individual examined shrimp, clustered within an unreported number of sampled aquariums from one seller and one claimed river source
Environment: Two hundred shrimp sampled across more than eight aquariums operated by the only ornamental-shrimp seller at one fish market in Purbalingga, Central Java, Indonesia
Endpoints: presence of reported organism group, host label, reported sex, gill or cephalothorax location.
What it can support: In this one-seller sample, the authors reported at least one of the three organism groups on 156 of 200 examined shrimp, including 78 of 100 under each market species label.
What it cannot support: Aquarium identities and sample counts, collection dates, host-species identification, sexing method, microscope magnification, diagnostic key, stain, image vouchers, molecular confirmation, examiner blinding, confidence intervals and the named statistical test were not reported. The two market labels were reportedly kept together, and the analysis did not account for aquarium clustering. No tissue damage, respiration, behaviour, growth, reproduction, mortality, transmission, treatment or causal disease endpoint was measured. The 78 percent result is not a trade-wide prevalence estimate, does not validate a home diagnosis, does not prove every detected organism was parasitic or harmful and does not establish a treatment indication.
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Controlled experiment
Direct Neocaridina evidence
Ostróżka et al., 2025. The European Zoological Journal 92(1): 876-895.
Source taxon as published: Neocaridina davidi
Study shape: A continuation using the 2024 nickel experiment scheme measured cell-cycle distribution, Ki67 proliferation, ATP and ADP/ATP, mitochondrial membrane potential, HSP70 and MnSOD in intestine and hepatopancreas. Five specimens represented each control or experimental group for each qualitative and quantitative method except Western blot. The prepublication CC BY deposit supplies the mitochondrial-potential and luminometry values plus microscopy and blot images.
Experimental unit: Individually exposed adult and its isolated organ or cell preparation, with five deposited values per group and organ for mitochondrial potential, ATP and ADP/ATP. The files do not provide specimen identifiers linking endpoints or establish whether the same five animals supplied each method; whether this was a new animal cohort independent of the 2024 publication was not stated.
Environment: Adult shrimp held individually in 0.5-litre cups at 21 C, pH 7 and GH 10 during nominal 3.63 mg Ni/L exposure for one or two weeks and one or two subsequent clean-water weeks
Endpoints: cell-cycle distribution, Ki67-positive cells, ATP concentration, ADP to ATP ratio, mitochondrial membrane potential, HSP70, MnSOD.
What it can support: The one high nominal nickel treatment produced organ- and time-associated changes in proliferation, energy and mitochondrial measures. Some measures returned toward control values during clean-water periods while others did not.
What it cannot support: This publication continues the prior nickel research program and uses its experimental scheme, so it is not evidence of an independent dose replication. Water concentration was not analytically verified, the control was the continuous culture rather than a separately described time-matched cup group, recovery groups were conditional on survival and only five values represented each deposited quantitative method group. The deposit does not link values across methods or prove independent reuse of animals. It did not measure whole-animal recovery, behaviour, reproduction or a safe household concentration.
Related source material: CC BY quantitative and image dataset
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Field observation
Direct Neocaridina evidence
Prati et al., 2025. BioInvasions Records 14(2): 403-419.
Source taxon as published: Neocaridina davidi; Ecytonucleospora hepatopenaei, Orthosomella lipae, Microsporidium sp. SRE01 and Microsporidium sp. SRE02 as named by the source
Study shape: Forty shrimp were dip-netted at each of three sites and preserved immediately in 96 percent ethanol. All were identified morphologically, measured, sexed or classified as immature, dissected, visually screened internally and tested molecularly. Host CO1 and microsporidian SSU rRNA amplicons were Sanger sequenced and placed with reference sequences in maximum-likelihood phylogenies with 1000 bootstrap replicates.
Experimental unit: Individual field-collected shrimp for morphology, internal visual screening and molecular detection; one date and three sites in two catchments for the field occurrence observations
Environment: Three freshwater sites in two catchments on La Reunion Island sampled on 15 April 2024 at 24.5 to 26.2 C and 76 to 268 microS/cm conductivity
Endpoints: host morphology and CO1 identity, sex and life-stage counts, carapace length, microsporidian PCR-positive count, sequence similarity and phylogenetic placement, site occurrence, one-day temperature and conductivity.
What it can support: Five of 120 sampled shrimp were PCR positive for four reported microsporidian sequence types: two Orthosomella lipae detections at the Salazie creek, and one Ecytonucleospora hepatopenaei, one Microsporidium sp. SRE01 and one Microsporidium sp. SRE02 detection at Bras Citronnier. None of the 40 Ravine Seche shrimp was positive. All 120 hosts shared one CO1 haplotype reported as 100 percent similar to the N. davidi reference set.
What it cannot support: This is a one-day field sample from three sites, not an aquarium or trade prevalence estimate. The paper reports no histology, tissue lesions, clinical signs, survival, experimental transmission, pathogenicity or treatment endpoint, and it did not test native co-occurring shrimp for parasites. Ethanol preservation prevented epibiont analysis. The single host CO1 haplotype cannot prove a single introduction, seller line, release event or movement route between catchments. Sequence similarity cannot establish parasite origin or that N. davidi introduced any organism. One Table 2 subgroup is internally inconsistent, listing 16 immatures while reporting two positives as 2 of 14, so subgroup percentages should not be repeated as settled denominators. The one-day site temperature and conductivity readings are not aquarium care limits, and zero of 40 at one site is not proof of absence.
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Controlled experiment
Adjacent biological evidence
Tiwari et al., 2025. Integrative and Comparative Biology 65(6): 1489-1497.
Source taxon as published: Dugesia tigrina, Dugesia dorotocephala and Lumbriculus variegatus; no Neocaridina
Study shape: Starved planaria encountered injured or uninjured short and long blackworms in thirty-four recorded behavioural trials.
Experimental unit: One planarian-blackworm behavioural trial
Environment: Thirty-five-millimetre freshwater Petri-dish predation trials near 21 C
Endpoints: grip and wrap behaviour, feeding success, prey injury and length context.
What it can support: The tested planaria used adhesion during blackworm capture, and prey injury changed the observed interaction.
What it cannot support: Blackworm capture does not establish predation on Neocaridina eggs, juveniles or adults, and body shape alone cannot identify the aquarium flatworm.
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Controlled experiment
Direct Neocaridina evidence
Ostróżka et al., 2024. The European Zoological Journal.
Source taxon as published: Neocaridina davidi
Study shape: The authors first screened nominal 0.09, 0.9 and 3.63 mg Ni/L, selected 3.63 mg Ni/L after observing about 50 percent mortality at 14 days, then compared one- and two-week exposures and one- or two-week clean-water periods. Whole-body nickel, midgut structure, reserves, reactive-oxygen and cell-death measures were assessed. The linked CC BY deposit supplies quantitative tables and microscopy files.
Experimental unit: Individually held adult for exposure, with animal, whole-body preparation or isolated organ used according to endpoint. The deposited TXRF table has three numbered samples per group, while reactive-oxygen tables contain uneven row counts and repeated analysis dates that do not identify every row as an independent animal.
Environment: Adult shrimp exposed individually to nickel chloride for one or two weeks, followed by one or two weeks in clean water at 21 C, pH 7 and GH 10
Endpoints: preliminary mortality, whole-body nickel, intestinal degeneration, hepatopancreatic change, reserve material, reactive oxygen species, autophagic signals, apoptosis and necrosis, clean-water response.
What it can support: At the selected high nominal nickel treatment, adults accumulated nickel and showed organ- and time-associated cellular damage; some measured changes persisted after one or two weeks in clean water.
What it cannot support: The approximately 50 percent 14-day mortality was a preliminary selection observation, not a reported formal LC50 estimate with uncertainty. The deposit measures nickel in whole-shrimp samples, not the exposure water, so 3.63 mg Ni/L remains nominal; the recovery groups necessarily contained survivors, controls remained in the continuous culture and the repository does not resolve whether every reactive-oxygen row is a biological unit rather than a repeat or batch observation. It does not establish whole-animal recovery, a household threshold, a copper or fertilizer limit, or a diagnosis from appearance.
Related source material: CC BY quantitative and image dataset
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Field observation
Direct Neocaridina evidence
Prati et al., 2024. Biological Invasions 26: 2499-2523.
Source taxon as published: Neocaridina davidi
Study shape: Feral occurrence, environmental conditions and intracellular parasite relationships were investigated.
Experimental unit: Field population, site and sampled animal, depending on endpoint
Environment: European feral populations with field sampling and molecular parasite work
Endpoints: feral occurrence, range expansion, microsporidian detection, environmental context.
What it can support: Feral populations and intracellular parasite associations exist in the sampled locations.
What it cannot support: One site value is not a care boundary, and field parasite detection is not a prevalence estimate for the aquarium trade.
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Morphology or taxonomy
Direct Neocaridina evidence
Hoitsy et al., 2023. Magyar Allatorvosok Lapja 145(6): 351-358.
Source taxon as published: Owner-submitted captive Neocaridina davidi from multiple breeders; host-identification method not reported
Study shape: Owners submitted 15 shrimp from different breeders after seeing moving white organisms on the day of introduction or within the following days. The shrimp were photographed, and associated live worms were collected into water drops, examined by stereomicroscopy and light microscopy, and identified from stated morphological references.
Experimental unit: Selected submitted shrimp for carrier observations and an unreported number of removed worms for morphological identification; five shrimp shared each of three holding aquaria, but no treatment or outcome comparison was assigned
Environment: Fifteen selected diagnostic submissions held five per approximately 18-litre aquarium at reported pH 6.8, conductivity 300 microsiemens and 25 C during examination
Endpoints: visible worm count and body region, worm length, live worm morphology, species identification, presence of worm eggs in the branchial chamber.
What it can support: All 15 selected shrimp carried visible opalescent-white moving worms, with 2 to 8 observed on the head, antennae or rostral region. Examined worms were reported as 0.8 to 2.0 mm long and identified as Scutariella japonica using paired anterior projections, two eyespots, internal reproductive anatomy and a posterior horseshoe- or heart-shaped sucker. Some worms contained eggs, and eggs were visible through the lateral carapace in the branchial chamber.
What it cannot support: This was a selected diagnostic case series, not a prevalence, transmission, quarantine-duration, water-quality or source-risk study. Host identity was not independently verified, source counts and clustering were not reported, the number of worms examined was omitted, and no molecular confirmation, voucher or image-accession record was stated. The study measured no respiration, gill lesion, molting failure, growth, reproduction, mortality, spontaneous clearance or treatment outcome. Harm statements were literature synthesis, not observations in these 15 shrimp. Fenbendazole was mentioned in one discussion sentence without product, dose, exposure, allocation, control, efficacy, recurrence or host-safety data. It cannot support a treatment recipe, a claim that every white worm is S. japonica or use of treatment response as diagnosis.
Related source material: University of Veterinary Medicine Budapest repository record and open PDF
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Controlled experiment
Direct Neocaridina evidence
Lima, 2023. UNESP undergraduate thesis, 22 pages.
Source taxon as published: Aquarium-shop Neocaridina davidi stock and aquarium-shop Biomphalaria glabrata; diagnostic methods and snail infection status were not reported
Study shape: Forty-two one-week-old shrimp were randomly selected and individually assigned as 14 animals per treatment to excess daily Sano S-PAK pellets, excised B. glabrata mantle, or an equal pellet-and-mantle offering. Animals were weighed every 14 days and sexed after 56 days.
Experimental unit: Individual 600 mL beaker containing one shrimp. The final analysis and figure contain 14 pellet, 13 snail-only and 13 mixed animals, but the loss of two of the 42 starters is not reconciled with the reported 100 percent survival.
Environment: Fifty-six-day laboratory feeding experiment at 25 C in individual 600 mL beakers with 1.6 g wet Java moss, daily partial water replacement and biofilm removal every 14 days
Endpoints: initial and final wet mass, mass gain, daily mass gain, specific growth rate, day-56 phenotypic sex, reported survival.
What it can support: Under this short individual-beaker protocol, the reported two-factor ANOVA detected no feeding-treatment main effect on mass gain and detected a sex main effect, with females gaining more mass than males.
What it cannot support: Failure to detect a treatment difference is not an equivalence or noninferiority test, and the small post-hoc sex groups were highly imbalanced. Food was offered in excess, but portion, consumed mass, leftovers, leaching and feed conversion were not measured. Snail mantle composition, pathogen screening, storage and water effects were not tested, and the snails had been raised on the same pellet used as the comparator. The abstract swaps the snail-only and mixed sex counts relative to the results, the stated 100 percent survival conflicts with 42 starters and 40 analyzed animals, and the mixed-female table mean of 0.335 g is impossible relative to the treatment final mass and plotted values. The paper reports no diet-by-sex interaction, exact treatment contrasts, power or raw data. It cannot establish that snail tissue is nutritionally complete, equivalent to formulated feed, pathogen-safe, humane to prepare, suitable as an infestation-control method or appropriate for routine home feeding.
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Morphology or taxonomy
Direct Neocaridina evidence
Maciaszek et al., 2023. Animals 13(10): 1616.
Source taxon as published: Neocaridina davidi
Study shape: One hundred shrimp from each of three locations within each source category were inspected individually in rostral, branchial, pereiopodal and pleopodal regions. Epibionts were isolated under clove-oil and ethanol sedation, identified morphologically at 500 to 1500x, and followed by an incompletely reported two-week quarantine observation. Prevalence models used source category and sex, microhabitat models used body region, and Spearman correlations assessed co-occurrence, egg production and molting.
Experimental unit: Individual examined shrimp nested within nine source locations and their transport bags; the reported individual-level GLMs do not describe source location or bag as a cluster or random effect
Environment: Nine hundred approximately 2.0 cm shrimp collected in November 2018 from three wild sites, three aquaculture-pond sources and three aquarium sources in Taiwan, then air-freighted in habitat water to Poland and inspected after a 10-hour acclimation in the transport-bag water
Endpoints: morphological identity and measurements, prevalence and mean intensity, co-occurrence, body-region distribution, egg-production association, molting association, observed cuticle traces and recolonization after molt.
What it can support: The authors distinguished Cladogonium kumaki, Monodiscus kumaki, Scutariella japonica and Holtodrilus truncatus, plus ciliates and rotifers. At least one recorded epibiont occurred on 688 of 900 shrimp and H. truncatus on 390 of 900. All four named taxa occurred in all four body regions with different distributions; H. truncatus was relatively associated with rostral and pereiopodal regions. Live H. truncatus and temnocephalids were observed leaving shed exuviae and seeking a host, sometimes recolonizing the nearby original shrimp.
What it cannot support: The source categories were observed, not assigned treatments, and are represented by only three locations each. Transport, ten-hour acclimation and microscopy handling may have changed organism abundance or location. Source-level clustering was not included in the reported GLMs, the sample was strongly female-skewed, and the sexing method was not stated. Morphological identification required microscopy and does not validate a phone-image diagnosis. Egg and molt analyses were correlational; the H. truncatus molt row reports rho -0.26 with p 0.847, an internally inconsistent combination for the surrounding interpretation. The quarantine allocation, denominator, water conditions, removal exposure and outcome measurements were not fully reported. Molting did not clear live worms, and the paper did not assign or compare a salt, medication or home-removal treatment. The sample is not prevalence for every seller, home colony or geographic trade route.
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Controlled experiment
Direct Neocaridina evidence
Melo, 2023. Doctoral thesis, Federal University of Minas Gerais.
Source taxon as published: Neocaridina davidi
Study shape: Separate total-ammonia and nitrite experiments used a control plus five treatments, two vessels per treatment and ten juveniles per vessel.
Experimental unit: Exposure vessel, not each shrimp
Environment: Renewed 96-hour acute laboratory exposures at about 25 C
Endpoints: 96-hour mortality, median lethal estimates, gill histology.
What it can support: Direct acute juvenile ammonia and nitrite exposure-response estimates with stated nitrogen reporting bases.
What it cannot support: A doctoral thesis with limited treatment replication does not provide chronic nitrate, chronic no-effect levels, transport limits or home targets.
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Morphology or taxonomy
Neocaridina identity evidence
Kakui and Komai, 2022. Aquatic Animals 2022: AA2022-1.
Source taxon as published: Neocaridina sp. aff. davidi
Study shape: Associated worms and host shrimp were examined for a regional record and taxonomic identification.
Experimental unit: Collected host or associated worm specimen
Environment: Field-collected host and associated worm specimens from Hokkaido, Japan
Endpoints: worm identification, host association, regional occurrence.
What it can support: A method-based record of Scutariella japonica associated with the source host identification.
What it cannot support: Occurrence does not establish aquarium prevalence, severity, treatment efficacy or identification from a distant white shape.
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Controlled experiment
Direct Neocaridina evidence
Kaya et al., 2022. Marine and Life Sciences 4(2): 146-151.
Source taxon as published: Neocaridina davidi, red cherry study stock
Study shape: A basal commercial feed, four feeds containing 1 to 4 g galacto-oligosaccharide per kg and four corresponding feeds also containing 1 mL Enterogermina per kg were assigned to three tanks each, with ten shrimp per tank for 30 days.
Experimental unit: Tank was the assigned treatment unit, with three tanks per diet and ten shrimp per tank; the published analysis did not state whether tank was modeled as the unit, and sex-specific groups were formed only after endpoint sexing
Environment: Twenty-seven 10-litre plastic tanks at 26 C with shelter, algae, twice-daily feeding to satiation and daily water measurements for 30 days
Endpoints: final body weight, weight gain, specific growth rate, feed conversion ratio, survival.
What it can support: Under this 30-day protocol, some galacto-oligosaccharide diets with or without the Bacillus clausii product had higher reported mass and growth metrics than the basal-feed control. The 2 g galacto-oligosaccharide plus 1 mL product per kg group had the highest table values for several sex-specific growth and feed-conversion outcomes.
What it cannot support: There was no probiotic-only group, so the design cannot isolate a Bacillus clausii main effect or demonstrate synergy. Finished-feed bacterial viability, additive retention, galacto-oligosaccharide composition and actual consumed ration were not verified; feeding was to satiation and no consumption correction was described. The statistical unit and endpoint sex grouping are unclear, initial sex-specific weights were repeated even though sex was not detectable at allocation, and there were only three tanks per diet for 30 days. Survival language is internally inconsistent. No microbiome, colonization, immune, disease-challenge, pathogen, reproduction, colour, body-composition, histology or long-term endpoint was measured. A human probiotic product in experimental feed is not an aquarium-water dose, bacterial starter, medicine or universal food recipe.
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Controlled experiment
Direct Neocaridina evidence
Ostróżka et al., 2022. Micron 154: 103162.
Source taxon as published: Neocaridina davidi
Study shape: Midgut tissues were compared across dimethoate exposure and recovery intervals.
Experimental unit: Animal and prepared midgut tissue
Environment: One to three weeks of laboratory exposure followed by clean-water recovery
Endpoints: midgut tissue damage, cellular regeneration, recovery interval.
What it can support: The tested exposure damaged midgut tissues and recovery responses differed among organs and intervals.
What it cannot support: It does not establish that a fixed clean-water period clears an aquarium or restores every biological endpoint.
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Field observation
Direct Neocaridina evidence
Schneider et al., 2022. Diseases of Aquatic Organisms 150: 125-130.
Source taxon as published: Neocaridina davidi; Enterocytozoon hepatopenaei and Microsporidium sp. EFB01 as named by the source
Study shape: Seventy-five ethanol-preserved adults were identified morphologically and screened for microsporidians by PCR from muscle tissue. Three randomly selected hosts received CO1 sequencing. Positive microsporidian amplicons were sequenced, compared with GenBank and placed in maximum-likelihood trees with 999 bootstrap replicates.
Experimental unit: Individual field-collected shrimp for PCR detection and carapace length; three randomly selected shrimp for host CO1 confirmation; one stream, one site and one sampling date for the population observation
Environment: One dip-net collection on 14 December 2021 from the Finkelbach, a German tributary of the River Erft, at 9 C, pH 7.65, dissolved oxygen 9.8 mg/L and reported conductivity 620 microS/cm
Endpoints: host identification, PCR-positive microsporidian count, sequence similarity, phylogenetic placement, sex, carapace length, visible epibiont and parasite examination.
What it can support: Four of 75 sampled shrimp were PCR positive for microsporidians. Three yielded sequences reported as 99.56 percent similar to Enterocytozoon hepatopenaei isolates, although only two were suitable for phylogenetic analysis; the fourth yielded the unresolved Microsporidium sp. EFB01 sequence. The three host CO1 sequences were 99.68 percent similar to one Taiwanese N. davidi reference.
What it cannot support: This is a one-date field detection from one German stream, not an aquarium-trade prevalence estimate or a transmission study. Only three hosts received molecular confirmation, and the paper does not report extraction blanks, PCR negative controls, product lengths, histology, tissue lesions, clinical signs, survival, transmission tests or treatment. PCR from muscle tissue cannot establish visible diagnostic characters or disease severity. The three E. hepatopenaei-positive shrimp were smaller than the 71 uninfected shrimp, but no supported inferential comparison was reported and three positives cannot establish growth suppression. Sequence similarity does not establish where either organism originated, that N. davidi introduced it, pathogenicity to native fauna, zoonotic risk or a home treatment.
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Field observation
Direct Neocaridina evidence
Bauer et al., 2021. Diseases of Aquatic Organisms 146: 107-115.
Source taxon as published: Neocaridina davidi
Study shape: Twenty submitted shrimp were assessed with clinical examination, wet mounts, microbiology, histology and sequencing of green pleopodal structures.
Experimental unit: Submitted shrimp, tissue section or algal sample, depending on method
Environment: Diagnostic examination of an imported commercial group during a multifactorial outbreak
Endpoints: organism morphology, sequence placement, cuticle penetration, co-occurring microbes, outbreak mortality history.
What it can support: The green organism was placed near Trentepohliales and rhizoids were observed penetrating cuticle into subcutaneous tissue.
What it cannot support: The multifactorial case does not assign every death to the alga, establish prevalence or let a green phone image confirm the organism.
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Controlled experiment
Direct Neocaridina evidence
Siregar et al., 2021. Antioxidants 10(3): 391.
Source taxon as published: Research-centre animals labelled Neocaridina denticulata; no diagnostic or molecular identification method reported
Study shape: A commercial 28.8 percent imidacloprid formulation was diluted to nominal 0.03125, 0.0625, 0.125, 0.25, 0.5 and 1 ppm for up to 96 hours. The paper describes 20 shrimp in one 200 mL plastic tank per treatment and 12 animals per group for movement, heartbeat and maxilliped-movement endpoints. A separate four-group series placed 24 animals per 500 mL tank, exposed three groups to nominal 1 ppm imidacloprid for 24 hours, then transferred them to fresh water or nominal 0.1 or 1 ppm acetylcholine for three days.
Experimental unit: Exposure tank for treatment assignment, apparently one tank per concentration or rescue group; individual shrimp were later measured in multiwell plates. Repeated video recordings and animal-level measurements do not create independent treatment-tank replication.
Environment: Small plastic exposure tanks using reverse-osmosis water at pH 7.0 to 7.5 after seven days of laboratory holding at 26.5 C
Endpoints: tracked distance and immobilization, image-derived heartbeat, maxilliped movement used as a gill-ventilation proxy, whole-animal oxidative-stress and energy-metabolism kit measurements, fresh-water and acetylcholine post-exposure trajectories, surrogate-protein molecular docking.
What it can support: Under this formulation and fasting protocol, imidacloprid-treated groups showed lower tracked movement across 24 to 72 hours, and higher nominal treatments had lower heartbeat and maxilliped-movement rates at 96 hours. The small whole-animal assay series reported no group differences in its selected oxidative-stress or energy-metabolism measures.
What it cannot support: Independent exposure tanks, random allocation, formulation-only controls and measured water concentrations were not reported. The text conflicts between 12 endpoint animals and 20 exposed animals per group, and treatment is confounded with the apparent single tank while animal measurements are analysed as replicates. Starvation changed heartbeat and maxilliped movement, acetylcholine itself changed the measured endpoints, and the rescue comparison also appears to use one tank per group. Docking used a snail acetylcholine-binding protein rather than a shrimp receptor and cannot prove the in vivo mechanism. The study does not validate acetylcholine as an antidote, identify imidacloprid from behaviour, establish recovery, chronic safety, a plant or flea-product rule, an aquarium dose or a universal threshold.
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Controlled experiment
Direct Neocaridina evidence
Hu et al., 2019. Environmental Pollution.
Source taxon as published: Neocaridina davidi
Study shape: Defined fenoxycarb and methoprene exposures were compared with controls.
Experimental unit: Exposure treatment culture unit
Environment: Chronic laboratory chemical exposure
Endpoints: body length, molting frequency, cuticle-related pathways, metabolism.
What it can support: The tested insecticides changed growth, molting and molecular outcomes under the exposure design.
What it cannot support: It does not establish that every pesticide acts the same way or identify a home exposure from behaviour or molting alone.
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Controlled experiment
Direct Neocaridina evidence
Włodarczyk et al., 2019. PeerJ 7: e7399.
Source taxon as published: Adult Neocaridina davidi obtained from local shrimp breeders
Study shape: Non-starved controls were compared with adults deprived of feed for 14 days and with groups re-fed for 4, 7 or 14 days after the same deprivation. DHE flow cytometry and confocal imaging assessed reactive-oxygen signals, immunofluorescence assessed MnSOD and a control-tissue western blot checked antibody specificity.
Experimental unit: Shrimp were isolated individually, but Table 1 lists 24 animals per flow-cytometry group and four per confocal method while the statistical section says assays used 5 to 6 samples in duplicate; the animal-to-assay sample mapping and any pooling are not fully reported
Environment: Adults were held in one 40-litre laboratory tank at 21 C, pH 7 and 10 dH, then isolated in shaded 250 mL containers with algae suppressed, excrement removed and 10 percent of the water replaced daily
Endpoints: percentage of DHE-labelled ROS-positive cells, qualitative tissue DHE signal, qualitative MnSOD immunofluorescence, antibody-specificity western blot.
What it can support: Under the tested algae-suppressed conditions, 14 days without feed increased the reported mean ROS-positive cell fraction from 2.8 to 13.2 percent in the hepatopancreas and from 1.3 to 12.7 percent in the intestine. Reported values declined toward control levels after 7 to 14 days of re-feeding, alongside weaker MnSOD immunofluorescence.
What it cannot support: The study tested one 14-day deprivation, not a weekly fast, and measured dissected-cell oxidative-stress markers rather than a survival, growth, molting, behaviour, reproduction or longevity benefit. Its isolated 250 mL containers deliberately suppressed algae and do not represent a conditioned community aquarium. Random allocation and blinding were not reported, sex was omitted from the analysis, assay sample mapping is incomplete, MnSOD treatment differences were not quantitatively measured, and the table, figure and statistical-method descriptions make the comparison-letter interpretation ambiguous. It cannot establish that routine fasting is beneficial, that missing one normal feeding is harmful, that biofilm is adequate, or that a specific re-feeding schedule is safe.
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Controlled experiment
Direct Neocaridina evidence
Włodarczyk, Student and Rost-Roszkowska, 2019. Canadian Journal of Zoology 97(4): 294-303.
Source taxon as published: Adult male and female Neocaridina davidi obtained from local shrimp breeders
Study shape: Adults were deprived of feed for 7, 14 or 21 days, and subsets were re-fed for 4, 7 or 14 days. TEM, TUNEL, LysoTracker and caspase-3 methods assessed intestine and hepatopancreas cells. Non-starved reference values and images were imported from a 2016 paper rather than produced as a concurrent control group.
Experimental unit: Shrimp were isolated individually, but Table 1 lists 10 TEM animals and 8 animals for each confocal method in most groups while quantitative Tables 2 and 3 report n = 5; animal reuse, pooling and the cell-to-animal analysis hierarchy are not resolved
Environment: Adults came from one 40-litre laboratory tank at 21 C, pH 7 and printed total hardness of 10 degrees d, then were isolated in shaded 250 mL containers with algae suppressed, excrement and exuviae removed and 10 percent of the water replaced daily
Endpoints: midgut cell ultrastructure, autophagosome-bearing cell percentage, TUNEL-labelled cell percentage, qualitative LysoTracker signal, qualitative caspase-3 signal, unquantified mortality during 21-day deprivation.
What it can support: Within this linked laboratory series, 7 and 14 days without feed were associated with more reported autophagosome-bearing midgut cells, and tissue descriptions after 14 days included fewer organelles and less reserve material. After re-feeding, qualitative tissue structure and the reported cellular markers moved toward the earlier non-starved reference.
What it cannot support: This paper reused the prior experimental procedure and imported its non-starved controls, so it is not independent replication and does not provide a concurrent baseline. Most animals deprived for 21 days reportedly died, but mortality counts, timing and survival analysis were absent; the paper says that group was not quantitatively analyzed even though Table 1 lists post-21-day TEM specimens. Re-feeding food amount, frequency and intake were not reported. Table 1 animal counts conflict with the n = 5 quantitative tables, the nested cell and animal hierarchy is unclear, random allocation and blinding were not reported, and the t tests omit comparison pairs, assumptions, exact p values and multiplicity handling. Table 3 prints zero means with positive standard deviations for nonnegative percentages, which is internally impossible. Caspase-3 and LysoTracker results were qualitative, marker validation was limited, and the TUNEL threshold was described as arbitrary. The study measured no growth, molting, behaviour, reproduction, longevity or whole-animal recovery and cannot establish a routine fast, safe missed-feeding interval, re-feeding recipe, home diagnosis or health benefit.
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Controlled experiment
Direct Neocaridina evidence
Sonakowska et al., 2017. PLOS ONE 12(3): e0173563.
Source taxon as published: Neocaridina davidi
Study shape: Midgut cell and mitochondrial responses were compared through starvation and refeeding periods.
Experimental unit: Animal and prepared midgut tissue
Environment: Laboratory starvation and refeeding experiment with tissue analysis
Endpoints: cell structure, mitochondrial potential, regeneration after refeeding.
What it can support: Starvation and refeeding changed method-confirmed midgut cell outcomes.
What it cannot support: It does not establish a routine fast, nutritional adequacy, a safe missed-feeding duration or a diagnosis from appearance.
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Morphology or taxonomy
Direct Neocaridina evidence
Ahn and Min, 2016. Journal of Species Research 5(3): 459-462.
Source taxon as published: Wild stream shrimp identified only as Neocaridina sp.; no host voucher, diagnostic method or molecular host identification reported
Study shape: The authors removed worms from preserved host exoskeletons under a stereomicroscope, described external and internal morphology, deposited examined material at Inha University and the National Institute of Biological Resources, and extracted whole-body DNA from one worm for partial mitochondrial COI amplification and sequencing.
Experimental unit: An unreported number of Neocaridina hosts and removed worms across three collection dates; one whole worm for the 674-base-pair COI sequence; three National Institute of Biological Resources voucher identifiers were listed
Environment: Hand-net collections from Andeok Valley, Jeju-do Province, South Korea on 23 June 2010, 23 August 2015 and 18 May 2016; hosts were preserved directly in 95 percent ethanol at the collection site
Endpoints: geographic and host occurrence, whole-body and jaw morphology, body length, voucher deposition, partial mitochondrial COI sequence.
What it can support: The record establishes morphologically identified H. truncatus on wild Neocaridina sp. at one South Korean stream across material dated in 2010, 2015 and 2016. Described characters included a transparent terete body under 2 mm, no trunk appendages, a posterior sucker about the diameter of the head region and paired jaws with a 7/7 dental formula. Three NIBR voucher identifiers and the first reported 674-base-pair COI sequence for the genus, GenBank KX683299, provide checkable taxonomic anchors.
What it cannot support: The paper does not state the number of hosts collected, hosts carrying worms, worms examined per date, worms per host, sampling effort or whether absence was assessed, so three dated collections do not estimate prevalence, persistence between dates or geographic range. The host remained Neocaridina sp. without a reported voucher or molecular identification. Although examined material was deposited at two institutions, only three NIBR identifiers were listed and the paper does not map individual morphology, image, sequence and voucher records. The single COI sequence created a barcode reference but was not compared with an existing conspecific sequence or used in a phylogenetic or species-delimitation analysis. Immediate ethanol preservation precluded live attachment, movement and host-response observations. The study measured no lesion, respiration, molting, growth, reproduction, mortality, transmission, origin, treatment, recurrence or host safety. It cannot establish N. davidi prevalence, aquarium-trade origin, harmlessness, a phone-image diagnosis, a quarantine duration or a treatment protocol.
Related source material: Korea Citation Index open article record and licensed PDF
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Controlled experiment
Adjacent biological evidence
Rivera-De la Parra, Sarma and Nandini, 2016. Journal of Limnology 75(s1).
Source taxon as published: Hydra predators and cladoceran, copepod and rotifer prey; no Neocaridina
Study shape: Hydra predation was compared across named zooplankton prey, and Hydra-conditioned water was tested against Daphnia demography.
Experimental unit: Predator-prey culture vessel
Environment: Laboratory freshwater predator-prey cultures
Endpoints: prey capture, prey susceptibility, Daphnia population growth.
What it can support: The tested Hydra captured several zooplankton taxa and affected one Daphnia population endpoint under the laboratory conditions.
What it cannot support: No Neocaridina life stage was tested, so the study does not prove that aquarium Hydra kill shrimplets or quantify colony risk.
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Morphology or taxonomy
Direct Neocaridina evidence
Sonakowska et al., 2016. PLOS ONE 11(2): e0147582.
Source taxon as published: Neocaridina heteropoda obtained from local breeders; no host-identification method reported
Study shape: Light, transmission-electron and confocal microscopy, acid-phosphatase and LysoTracker staining, TUNEL labeling and JC-1 flow cytometry were used to describe autophagy, apoptosis, necrosis and mitochondrial potential in intestine and hepatopancreas tissue from method-specific adult groups. No external stressor or disease treatment was assigned.
Experimental unit: Prepared tissue or cell suspension from method-specific adult specimens: 15 females and 15 males for general microscopy, three adults for quantitative necrosis and autophagy cell counts, two for TEM acid-phosphatase staining, 10 females and 10 males for cryosections, five pooled within each organ suspension preparation and seven for JC-1 flow cytometry; the TUNEL animal denominator was not stated
Environment: Adults maintained in 30 L laboratory aquaria at 24 C, pH 7 and 15 degrees German total hardness and fed a commercial freshwater-shrimp food
Endpoints: cell-type and organ location, autophagic structures, TUNEL-positive apoptotic cells, necrotic-cell morphology, mitochondrial membrane potential, sex comparison.
What it can support: Under the described laboratory history and without an assigned external stressor, the examined adult tissues contained autophagic, apoptotic and occasional necrotic cells. Autophagy was observed in mature intestine D cells and hepatopancreas B and F cells, apoptosis in anterior-intestine D cells and proximal hepatopancreas B and F cells, while regenerative cells showed none of the three reported processes. The study did not report a statistically significant intestine-versus-hepatopancreas difference in cells with depolarized mitochondria.
What it cannot support: This is destructive cellular and tissue evidence, not a visible home diagnosis or a disease experiment. Breeder source, sex and adult status were reported, but host identification, age, molt stage, reproductive state, tank count, animals per tank, acclimation duration, health screening, water-test methods and the TUNEL animal denominator were not. Method groups differed and their overlap was not reported. Cell counts were nested within only three adults for the necrosis and autophagy percentages, TUNEL used an arbitrary fluorescence threshold and sex-specific values or tests were not shown. Table 1 prints 938 autophagic cells from 231 total for intestine specimen 3 even though the displayed 40.26 percent implies 93. The results do not establish a gross sign, pathogen, cause of illness, welfare state, treatment target, aquarium optimum or safe exposure, and the maintained 24 C, pH 7 and 15 degree hardness history was not an assigned comparison.
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System observation or method study
Direct Neocaridina evidence
Tanaka, Wada and Hamasaki, 2016. Zoological Science 33(2): 154-161.
Source taxon as published: Holtodrilus truncatus; field hosts included Paratya compressa, Caridina leucosticta, Caridina multidentata, Caridina typus and Neocaridina denticulata
Study shape: The field survey standardized dip-net sampling at ten waterside areas per site and repeated one river seasonally. The salinity study exposed 24, 20, 30, 25 and 12 host-removed worms to nominal 0, 0.5, 1, 1.5 and 3 percent salinity without food, with observation intervals that shortened as salinity rose. Host-choice tests placed one worm with a size-matched pair of Paratya compressa, Caridina leucosticta or Caridina multidentata for 40, 36 and 39 pairwise replicates.
Experimental unit: Field host shrimp for occurrence endpoints; an incompletely reported small plastic tank holding two or five isolated worms for salinity exposure; and one worm with one two-shrimp species pair for each host-choice replicate
Environment: Field collections at one to eight sites in each of 26 rivers on the Kii Peninsula, Japan, plus 22 to 27 C freshwater laboratory salinity and two-host choice tests
Endpoints: river and host occurrence, prevalence and intensity on Paratya compressa, host body location, Paratya compressa egg number association, isolated-worm attachment-defined survival, selected host in pairwise choice.
What it can support: Holtodrilus truncatus occurred in 10 of 26 surveyed rivers and on five atyid host species. The only two examined N. denticulata were both recorded with the worm, which establishes occurrence but not prevalence. Isolated-worm survival declined with increasing nominal salinity, including significantly lower survival at 0.5 than 0 percent. In pairwise tests that excluded Neocaridina, worms selected P. compressa over both Caridina species and C. leucosticta over C. multidentata. Cohabitation was not associated with egg number in the measured P. compressa model.
What it cannot support: The direct Neocaridina result is two field animals, not a representative sample. No Neocaridina entered the salinity, host-choice or egg-number analyses. Worms were removed after the host carapace was detached, so salinity was tested on isolated organisms rather than attached worms or living host shrimp. The work measured no treatment efficacy on a host, shrimp survival, sublethal host effect, recurrence or safety margin. Worms shared exposure containers in groups of two or five, the number of independent tanks was not reported, checking frequency differed by concentration, no food was provided, and floating or failure to attach was classified as death. Nominal salinity tolerance cannot validate a salt dip, 30-second exposure, host-safe dose or Neocaridina host preference.
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System observation or method study
Direct Neocaridina evidence
Cheung et al., 2015. Marine Biotechnology 17(6): 811-819.
Source taxon as published: Laboratory red-patched Neocaridina denticulata denticulata linked to the Kenny et al. stock; the commercial foundation stock had no reported voucher, morphology or molecular host-identification method
Study shape: Three adult females at each of three visually assigned ovarian stages were sampled cross-sectionally. Foregut, intestine including midgut and hindgut, and hepatopancreas were dissected from every female, producing 27 individually sequenced 16S V1-V3 libraries. PCR reactions were run in triplicate and pooled within each library. Ion Torrent reads were processed in QIIME 1.8, clustered into 97 percent OTUs, assigned against Greengenes at 0.5 confidence, rarefied to 14,000 reads and compared with alpha-diversity indices, UniFrac PCoA and UPGMA trees.
Experimental unit: Individual female for ovarian-stage comparisons, with three females per stage and three linked tissue libraries per female. Triplicate PCR reactions were technical replicates pooled within a library, not additional animals. The paper did not report a repeated-measures model for tissues from the same female.
Environment: Adult females from a multigeneration recirculating freshwater laboratory stock at approximately 25 C; all were starved for two days before dissection
Endpoints: quality-filtered 16S read count, 97 percent OTU richness, Chao1 richness, Shannon diversity, phylogenetic diversity, weighted and unweighted UniFrac clustering, relative read abundance by bacterial taxon, SRA accession SRR1735538.
What it can support: Within this nine-female stock after two days of starvation, early-stage foregut and intestine libraries had lower reported alpha diversity than later-stage libraries, hepatopancreas libraries generally differed from gut libraries, and more than 80 percent of early-stage gut reads were assigned to one Coxiella-type OTU. The study provides a public 16S survey associated with tissue region and visually assigned ovarian stage.
What it cannot support: This was a small cross-sectional association, not the same females followed through ovarian development and not an assigned microbiota or reproductive treatment. Stage could be confounded with age, time, individual and egg-carrying status. Relative read abundance is compositional and does not measure absolute bacterial load, prevalence among shrimp or colonization. A short 16S Coxiella-type assignment at a low classification threshold is not a species or strain identification, Coxiella burnetii diagnosis, cultured isolate, pathogen finding or probiotic. The paper reported t tests without specifying exact contrasts, pairing, tails, variance handling, test statistics, degrees of freedom or multiplicity correction, and reported no formal group-level beta-diversity test. No extraction blank, PCR negative control, mock community, random selection or blinding was described. The paper did not test bacterial function, microbial transfer, probiotic or antibiotic exposure, fertility, spawning, hatch, offspring, growth, survival, disease, immune response, feed treatment or normal fed-tank baseline. It cannot show that microbes cause ovarian maturation, that maturation causes the microbiota difference or that changing aquarium bacteria improves breeding or health.
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Controlled experiment
Direct Neocaridina evidence
Li et al., 2015. Environmental Science 36(2): 727-735.
Source taxon as published: Wild animals from vegetation around Dianshan Lake labelled Neocaridina denticulate; no diagnostic or molecular identification method reported
Study shape: Separate five-day semi-static series used nominal copper-ion treatments of 0.086, 0.172, 0.344 and 0.688 mg/L and nominal chlorpyrifos treatments of 0.0015, 0.0030, 0.0060 and 0.0120 micrograms/L. Each concentration was described as three parallel 2-litre beakers with 20 shrimp per beaker, daily solution renewal, no feeding and blank-water and chlorpyrifos vehicle controls. Three shrimp per group were destructively sampled on each day for muscle homogenate assays.
Experimental unit: The two-litre exposure beaker, described as three parallel beakers per concentration. The methods do not state how the three shrimp sampled at each time were distributed among or pooled across those beakers, although results are described as means of three parallel groups.
Environment: Two-litre beakers in temperature-controlled water baths using aerated tap water at 23 +/- 1 C, pH 7.2 to 7.7, hardness about 205.95 mg/L as CaCO3 and dissolved oxygen above 5.80 mg/L
Endpoints: five-day mortality observation, muscle protein concentration, muscle malondialdehyde content, muscle total superoxide dismutase activity, muscle acetylcholinesterase activity.
What it can support: Under the tested semi-static protocols, nominal copper-ion and chlorpyrifos concentration series were associated with time-varying muscle protein, lipid-peroxidation and enzyme-activity measurements. No deaths were reported through five days in these sublethal treatment groups.
What it cannot support: The wild-source taxon was not independently verified as N. davidi, concentrations were nominal rather than analytically measured, animals were fasted for five days and sampling-to-beaker mapping is unclear. The paper cites 96-hour LC50 values from earlier work to select its fractions but does not provide that acute experiment, uncertainty or full analysis, so those values are not results of this five-day series. Repeated t tests and one-way ANOVAs across concentration-by-day contrasts had no reported multiplicity or repeated-time model, and the text contains concentration-label inconsistencies. Muscle biomarkers do not establish a no-effect level, chronic safety, diagnosis, antidote, aquarium dose or universal copper or pesticide threshold.
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Review or synthesis
Broader ornamental-shrimp review
Mykles and Hui, 2015. Integrative and Comparative Biology 55(5): 891-897.
Source taxon as published: Policy review comparing Neocaridina denticulata with the crayfish Procambarus clarkii and Cherax quadricarinatus, with additional inferences from other crustacean models
Study shape: The authors defined desired traits for a decapod model, compared three freshwater candidates, summarized selected Neocaridina life-history, culture, anatomy and genomic claims from earlier publications, recommended N. denticulata, and proposed obtaining a complete genome and developing tissue-specific gene-control and mutant-screening methods.
Experimental unit: Published source or narrative claim; the paper reported no new animals, aquaria, assigned treatments, measurements or statistical analysis and did not describe a systematic search, eligibility criteria or risk-of-bias assessment.
Environment: No new animal experiment or culture trial; narrative synthesis and recommendation arising from a 2015 comparative Pancrustacea symposium workshop
Endpoints: model-organism selection criteria, comparative recommendation, summary of cited life-history and culture claims, summary of draft-genome status, proposed functional-genomics next steps.
What it can support: The paper documents that two authors recommended N. denticulata as a decapod functional-genomics model in 2015 based on its small space requirement, commercial availability, transparent cuticle, cited life-history traits and then-available preliminary genome. It explicitly identified a complete genome and reliable gene-expression manipulation as future work.
What it cannot support: This policy review is not a new replication of any cited experiment and is not a systematic review. Its pH 6.5 to 8.0, temperature up to 30 C, preferred 22 to 25 C and pH 7.0 to 7.5, 15 to 16 day intermolt, 20 to 30 egg, approximately 30 day hatch and 4 to 6 month maturity statements combine older sources, populations and methods; they are not one standardized aquarium comparison or validated species-wide optimum. The broad statement that N. denticulata is resistant to bacterial infection does not establish general disease resistance, a pathogen-free line, treatment or immune supplement. The cited DPrP bacterial-challenge paper remains full-text-needed in this registry, and the companion crayfish-plague citation studied a related published species against one oomycete pathogen. The preliminary genome was explicitly incomplete, and proposed transgenesis, reporter expression, mutation screening and targeted gene control were future possibilities rather than demonstrated Neocaridina methods in this paper. The review does not validate a care range, breeding calendar, stocking rule, disease guarantee, commercial-line assay or consumer genetic intervention.
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Morphology or taxonomy
Direct Neocaridina evidence
Sonakowska et al., 2015. PLOS ONE 10(5): e0126900.
Source taxon as published: Neocaridina heteropoda
Study shape: Digestive structures were examined with microtomography, light microscopy and electron microscopy.
Experimental unit: Prepared animal and tissue specimen
Environment: Laboratory microscopy and microtomography
Endpoints: intestine structure, hepatopancreas structure, cell ultrastructure.
What it can support: Method-confirmed internal digestive anatomy in the source taxon.
What it cannot support: An ordinary tank photograph cannot identify these internal organs, function or disease from colour alone.
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System observation or method study
Direct Neocaridina evidence
Niwa et al., 2014. Central European Journal of Biology 9(1): 80-85.
Source taxon as published: Wild Sugo River shrimp identified only as Neocaridina spp.; the catch could include native N. denticulata denticulata and imported Neocaridina lineages
Study shape: The authors collected 271 host shrimp across nine years, mapped H. truncatus body locations on 141 hosts in 2011, measured 23 cocoons, observed 15 host-removed worms from 14 shrimp in no-added-food survival containers, and placed five removed worms sequentially with a reported pool of 28 potential host shrimp across several taxa.
Experimental unit: Individual host for mapped attachment location; individual cocoon for diameter; nominally individual host-removed worm, although only 13 of 15 worms receive a reported container and survival range; and an unresolved sequence of five worms exposed one host at a time in the host-exchange demonstration
Environment: Field collections at three Sugo River stations from 2003 to 2011 plus laboratory observations in source river water, aerated aquaria, individual bottles, dishes and 100 mL vials
Endpoints: host body location, cocoon diameter and embryo count, observed hatching and gill-chamber entry, days survived after host removal, attachment or predation during host exchange, movement from deteriorating to surviving captive hosts.
What it can support: Among 141 mapped wild hosts, reported H. truncatus locations were 55.3 percent between the first pleopod and fifth pereiopod, 17.0 percent on the carapace, 15.6 percent at the eye base, 8.5 percent on the antennule and 3.6 percent around the egg mass. Twenty-three cocoons measured 0.58 to 0.76 mm, and the largest observed cocoon contained 14 developing worms. Ten worms in 100 mL vials survived 7 to 46 days and three in dishes survived 12 to 21 days after host removal without added food. In the host-exchange observation, all five supplied worms attached to Neocaridina hosts within three hours.
What it cannot support: Host shrimp were resolved only to Neocaridina spp. in a river where native and introduced lineages were discussed, so the findings are not species-specific N. davidi evidence. The 271 total catch, 152 animals mentioned in the attachment procedure and 141 mapped hosts are not fully reconciled, station and date denominators are missing and the location percentages do not state whether each host contributed one location or all worms were counted. No molecular confirmation of host or worm identity was reported in this article. The survival containers differed in volume, light and temperature, two of 15 worms are absent from the reported ranges, ages were unknown, water was not renewed or analytically described, death criteria were not stated and there was no fed or host-present control. The five-worm host exchange omitted exposure order, reuse mapping, independent replication, container details and complete outcomes for 28 potential hosts. Disappearance under an opaque crayfish carapace was interpreted as migration rather than directly confirmed. Claims that egg-bearing hosts hatched healthy young had no defined denominator, comparison or follow-up. Observed migration from deteriorating hosts does not establish transmission probability or a quarantine duration. The study measured no controlled shrimp lesion, respiration, growth, reproduction, mortality causation, treatment efficacy, recurrence or host safety and cannot validate predatory tankmates, forced host deterioration, worm collection, medication or a fixed isolation period.
Related source material: German National Library open full-text copy
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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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Morphology or taxonomy
Adjacent biological evidence
Van Steenkiste et al., 2013. PLOS ONE 8(3): e59917.
Source taxon as published: Dalytyphloplanida flatworms, not Neocaridina
Study shape: Complete 18S and partial 28S rDNA sequences were analysed with maximum-likelihood and Bayesian phylogenetic methods.
Experimental unit: Taxon sequence or specimen represented in the phylogeny
Environment: Molecular phylogenetic analysis across marine, freshwater and symbiotic flatworms
Endpoints: phylogenetic placement, freshwater transition reconstruction, symbiotic-lifestyle reconstruction.
What it can support: Flatworm lineages and life strategies cannot be inferred from a generic worm-like aquarium silhouette alone.
What it cannot support: The phylogeny does not identify a phone-video organism, demonstrate predation or determine whether an observed flatworm threatens shrimp.
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Controlled experiment
Direct Neocaridina evidence
Wang et al., 2010. Israeli Journal of Aquaculture - Bamidgeh 62(2): 78-84.
Source taxon as published: Wild-collected Neocaridina heteropoda from Baiyangdian Lake; no diagnostic identification method reported
Study shape: Nine hundred shrimp were assigned at 50 per tank to six manganese-sulfate supplementation levels, with three tanks per diet. The measured complete-diet manganese concentrations were 2.79, 22.57, 42.55, 62.69, 82.59 and 102.45 micrograms per gram. Shrimp received feed at a stated daily rate of 5 percent body weight for 30 days.
Experimental unit: Tank for the six assigned diets, with three tanks per diet in one shared-water recirculating system. Figure 1 reports n = 6 for biochemical assays, but the paper does not map those samples or pooled hemolymph back to individual shrimp and tank replicates or state whether tank entered the analysis.
Environment: Thirty-day freshwater recirculating-system feeding experiment at 25 +/- 1 C and pH 8.0, with all tanks connected to one reservoir and reported water manganese of 1.00 +/- 0.02 micrograms per litre in the abstract but micrograms per gram in the methods
Endpoints: percentage length gain, percentage weight gain, hemolymph superoxide-anion assay, muscle SOD activity, muscle GPX activity, muscle CAT activity.
What it can support: Under this complete-feed formulation and 30-day system, every manganese-supplemented group had higher reported length and weight gain than the 2.79-microgram-per-gram control diet. The 62.69-microgram-per-gram measured diet had the highest reported growth percentages and antioxidant-enzyme activities and the lowest reported superoxide-anion assay value.
What it cannot support: The experiment does not establish a universal dietary requirement or deficiency threshold. The control was not manganese-free, the basal mineral premix already listed manganese sulfate, and the diets differed only within one formulation and short culture history. The paper reports neither starting-to-ending size values nor a growth formula, feed intake, feed conversion, survival, tissue manganese, deficiency signs, molting, reproduction or long-term safety. Wild-source identity, sex and life stage were not resolved. Sample-to-tank mapping for the n = 6 biochemical results and pooled hemolymph is unclear, and the water manganese unit conflicts between abstract and methods. A peak response at one tested concentration without dose-response modeling does not validate adding manganese to aquarium water, mixing a supplement into finished food or using antioxidant-enzyme activity as proof of better health.
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