Evidence map›Paper›PMID 40022439›Full record

ArticleReproduction in domestic animals = Zuchthygiene2025

Application of Staining Methods to Compare Chromatin Condensation in Fresh and Freeze-Thawed Dog Semen.

Elisa-Marie Laux, Abbas Farshad, Axel Wehrend, Mohamad Eid Hammadeh

Abstract readComparative Study
In one paragraph

Article in Reproduction in domestic animals = Zuchthygiene, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 4 papers.

0numbers the graph read from it
0cells of the map it votes in
4citing papers in PubMed
–field-weighted citation impact
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

4 citing papers in PubMed.

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4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

4 authors.

Elisa-Marie LauxVeterinary Clinic for Reproductive Medicine and Neonatology, Justus-Liebig-University of Giessen, Giessen, Germany.
Abbas FarshadVeterinary Clinic for Reproductive Medicine and Neonatology, Justus-Liebig-University of Giessen, Giessen, Germany.ORCID https://orcid.org/0000-0002-1502-7797
Axel WehrendVeterinary Clinic for Reproductive Medicine and Neonatology, Justus-Liebig-University of Giessen, Giessen, Germany.
Mohamad Eid HammadehObstetrics & Gynecology, Department of Biochemistry and Molecular Biology of Reproductive Medicine Laboratory, University of Saarlandes, Hombur/Saar, Germany.

Funding

No grant is acknowledged in the PubMed record.

6 · The paper itself

Abstract

Various staining techniques have been used for canine sperm analysis, but direct comparisons using identical semen samples are lacking. This study aimed to assess the efficiency, time requirements and cost-effectiveness of different staining techniques: aniline blue, toluidine blue, acridine orange (AcO), chromomycin A3 (CMA3) and terminal deoxynucleotidyl transferase d-UTP nick-end labeling (TUNEL). Forty semen samples (20 fresh and 20 frozen-thawed) were used to assess chromatin condensation. Significant differences (p < 0.01) were found using two-factor repeated measures variance analysis. Aniline blue staining differed significantly (p < 0.01) from toluidine blue, AcO and CMA3 staining. A significant difference (p < 0.05) was observed between AcO and TUNEL staining for fresh sperm, with no significant differences between TUNEL and other methods. Correlations in fresh sperm samples showed r = 0.567 between AcO and aniline blue, r = 0.645 between AcO and CMA3, and correlations of 0.455 and 0.557 for aniline blue - toluidine blue and aniline blue - TUNEL, respectively. For frozen samples, significant differences were found between aniline blue and toluidine blue and AcO tests (p < 0.05), and between CMA3 and TUNEL staining (p < 0.01). Correlations in frozen samples showed r = 0.582 between AcO and aniline blue, r = 0.752 between AcO and CMA3, r = 0.698 between toluidine blue and aniline blue, and r = 0.536 between CMA3 and aniline blue. A minimal association was found between standard semen analysis and chromatin analysis. In conclusion, toluidine blue is effective for light microscopy staining, while CMA3 is recommended for fluorescence microscopy due to its simplicity, rapidity and cost-effectiveness.

Indexed as

ChromatinCryopreservationSemenSemen AnalysisSemen PreservationStaining and LabelingAcridine OrangeAniline CompoundsAnimalsDogsIn Situ Nick-End LabelingMaleSpermatozoaTolonium ChlorideAcridine Orangeaniline blueAniline CompoundsChromatinTolonium Chloridecaninechromatincondensationcryopreservationsperm

Identifiers

PMID40022439
PMCPMC11871417

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Registered trials

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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.