Data and code from: Characterizing neuroma-like formation after tail docking in sheep
Data files
Apr 20, 2026 version files 1.05 GB
-
Raw_images_251123.zip
674.09 MB
-
Raw_images_used_in_figures_251123.zip
376.11 MB
-
README.md
17.99 KB
-
Sheep-tail-docking-neuroma_analysis_RVS_251218.nb.html
796.38 KB
-
Sheep-tail-docking-neuroma-data_RVS_251218.xlsx
11.04 KB
Abstract
Tail docking is a routine husbandry procedure in lambs that causes acute pain. Studies of tail docking performed between 4 and 8 weeks of age have also reported neuromas in the tail stump, which are characterized by aberrant nerve proliferation and fibrotic scarring and represent a potential source of chronic pain. However, it remains unclear whether similar lesions develop after rubber ring docking in the first week of life, the age at which this method is commonly recommended. This study examined the histological characteristics of tail stumps in lambs docked with a constrictive rubber ring at 1-2 days of age compared with undocked controls. Eighteen Polypay female lambs (8 docked, 10 undocked) were slaughtered at 7.5 to 8 months of age and their tails collected and decalcified for histology and immunohistochemistry. Tissues were evaluated for abnormalities characteristic of neuromas followed by blinded semi-quantitative scoring of key histologic features on a 5-point severity scale. Neuroma-like axon proliferation and fibrosis ranged from mild to severe and were significantly more severe in docked than undocked tails. In addition to neuromatous lesions in the soft tissue, axon proliferation was particularly prominent in the periosteum of the docked vertebral bone. Masson’s trichrome staining showed that neural proliferation was accompanied by fibrosis, and immunohistochemistry confirmed that lesions included proliferation of both axons and Schwann cells, characteristic of traumatic neuromas. These findings provide evidence that tail docking with a rubber ring, even when performed at an early age, causes neuroma-like lesions, which may represent a source of chronic pain in sheep.
This repository contains the data and R script used to conduct the descriptive and statistical analyses and create the figures referenced in the paper of the same name.
📄 Study Overview
This study aimed to:
- Characterize the histomorphology of the tail stump in lambs docked with a rubber ring at 1-2 days of age and examined at slaughter (7.5-8 months of age), compared to age-matched undocked controls
- Compare the degree of neuroma formation in docked sheep vs. undocked sheep using semi-quantitative methods
Subjects:
Eighteen Polypay ewe lambs were randomly assigned to one of two docking treatments:
- Docked using the rubber ring method at 1-2 days of age (n = 8)
- Undocked controls (n = 10)
Tissue processing and histology:
All ewes were euthanized at 7.8 ± 0.2 months of age (mean ± SD) and their tails collected and decalcified for histology. FFPE sections were stained with H&E and trichrome, and immunohistochemistry was performed using stains for s100, calcitonin gene related peptide (CGRP), and neurofilament heavy chain (NFH).
Scoring and analysis:
Tissue samples were evaluated in a blinded manner for abnormalities characteristic of neuromas:
- Fibrosis
- Axon sprouting
Tissues were also evaluated for two additional characteristics noted during tissue processing: - Periosteal thickening
- Periosteal axon proliferation
Each tissue sample received a severity score ranging from 0 (absent) to 4 (severe) for the above characteristics. Tissue samples were unblinded and scores were compared between the two treatment groups.
📁 Files and Descriptions
Sheep-tail-docking-neuroma_analysis_RVS_251218.nb.html
- Description: R script to organize and visualize scoring data used to compare neuromas in docked vs. undocked sheep.
- Input:
Sheep-tail-docking-neuroma-data_RVS_251218 - Output:
Figure_5.tif,Figure_6.tif - Use: Open in any web browser or in RStudio (File → Open File)
Sheep-tail-docking-neuroma-data_RVS_251218.xlsx
- Description: Excel file containing raw scoring data from the blinded images.
| Variable | Description |
|---|---|
sheep_id |
Unique ID number assigned to each sheep at the beginning of the study. |
group |
Treatment group for each sheep (docked or undocked). |
DOB |
Date of birth of the sheep. |
dam_id |
Unique ID number of the ewe (dam) that gave birth to the sheep. |
birthweight_kg |
Birth weight of the sheep, in kilograms. |
slaughter_date |
Date on which the sheep was slaughtered for tissue collection. |
nerve_blind_id |
Unique blinded ID assigned to each histological image of peripheral nerve ("nerve bundle") selected for scoring, such that the sheep’s identity and treatment group were unknown to the observers. |
nerve_bundle_fibrosis_skp |
Fibrosis score for peripheral nerve tissue assigned by observer SKP. |
nerve_bundle_fibrosis_lkc |
Fibrosis score for peripheral nerve tissue assigned by observer LKC. |
nerve_bundle_axon_sprouting_skp |
Axon sprouting score for peripheral nerve tissue assigned by observer SKP. |
nerve_bundle_axon_sprouting_lkc |
Axon sprouting score for peripheral nerve tissue assigned by observer LKC. |
periosteal_blind_id |
Unique blinded ID assigned to each histological image of periosteum selected for scoring, such that the sheep’s identity and treatment group were unknown to the observers. |
periosteal_thickening_skp |
Periosteal thickening score assigned by observer SKP. |
periosteal_thickening_lkc |
Periosteal thickening score assigned by observer LKC. |
periosteal_axon_proliferation_skp |
Periosteal axon proliferation score assigned by observer SKP. |
periosteal_axon_proliferation_lkc |
Periosteal axon proliferation score assigned by observer LKC. |
Raw_images_251123.zip
- Description: Raw images of peripheral nerve and periosteum used in the study.
| File name | Description |
|---|---|
22324sp2_nerve_20x1.tif |
Image of peripheral nerve used for blinded scoring |
22326sp2_nerve_20x1.tif |
Image of peripheral nerve used for blinded scoring |
22326sp2_nerve_20x2.tif |
Image of peripheral nerve used for blinded scoring |
22332sp1_nerve_20x1.tif |
Image of peripheral nerve used for blinded scoring |
22338sp2_nerve_20x1.tif |
Image of peripheral nerve used for blinded scoring |
22339sp2_nerve_20x1.tif |
Image of peripheral nerve used for blinded scoring |
22355sp4_nerve_20x1.tif |
Image of peripheral nerve used for blinded scoring |
22356sp2_nerve_20x1.tif |
Image of peripheral nerve used for blinded scoring |
22369sp1a_nerve_20x1.tif |
Image of peripheral nerve used for blinded scoring |
22370sp1_nerve_20x1.tif |
Image of peripheral nerve used for blinded scoring |
22372sp1_nerve_20x1.tif |
Image of peripheral nerve used for blinded scoring |
22422sp1a_nerve_20x1.tif |
Image of peripheral nerve used for blinded scoring |
22422sp1b_nerve_20x1.tif |
Image of peripheral nerve used for blinded scoring |
22423sp1_nerve_20x1.tif |
Image of peripheral nerve used for blinded scoring |
22434sp2_nerve_20x1.tif |
Image of peripheral nerve used for blinded scoring |
22434sp2_nerve_20x2.tif |
Image of peripheral nerve used for blinded scoring |
22435sp1_nerve_20x1.tif |
Image of peripheral nerve used for blinded scoring |
22435sp1_nerve_20x2.tif |
Image of peripheral nerve used for blinded scoring |
22454sp1_nerve_20x1.tif |
Image of peripheral nerve used for blinded scoring |
22455sp2_nerve_20x1.tif |
Image of peripheral nerve used for blinded scoring |
22455sp2_nerve_20x2.tif |
Image of peripheral nerve used for blinded scoring |
22502sp1_nerve_20x1.tif |
Image of peripheral nerve used for blinded scoring |
22503sp1_nerve_20x1.tif |
Image of peripheral nerve used for blinded scoring |
22503sp1_nerve_20x2.tif |
Image of peripheral nerve used for blinded scoring |
22324sp1_perio_1.tif |
Image of periosteum used for blinded scoring |
22324sp2_perio_2.tif |
Image of periosteum used for blinded scoring |
22326sp2_perio_1.tif |
Image of periosteum used for blinded scoring |
22332sp2_perio_1.tif |
Image of periosteum used for blinded scoring |
22338sp1_perio_1.tif |
Image of periosteum used for blinded scoring |
22339sp2_perio_1.tif |
Image of periosteum used for blinded scoring |
22355sp1_perio_1.tif |
Image of periosteum used for blinded scoring |
22356sp1_perio_1.tif |
Image of periosteum used for blinded scoring |
22369sp1a_perio_1.tif |
Image of periosteum used for blinded scoring |
22369sp1a_perio_2.tif |
Image of periosteum used for blinded scoring |
22370sp2_perio_1.tif |
Image of periosteum used for blinded scoring |
22372sp1_perio_1.tif |
Image of periosteum used for blinded scoring |
22422sp1b_perio_1.tif |
Image of periosteum used for blinded scoring |
22423sp1_perio_1.tif |
Image of periosteum used for blinded scoring |
22434sp2_perio_1.tif |
Image of periosteum used for blinded scoring |
22435sp1_perio_1.tif |
Image of periosteum used for blinded scoring |
22454sp1_perio_2.tif |
Image of periosteum used for blinded scoring |
22454sp2_perio_1.tif |
Image of periosteum used for blinded scoring |
22454sp2_perio_3.tif |
Image of periosteum used for blinded scoring |
22455sp2_perio_1.tif |
Image of periosteum used for blinded scoring |
22502sp1_perio_1.tif |
Image of periosteum used for blinded scoring |
22502sp1_perio_2.tif |
Image of periosteum used for blinded scoring |
22503sp1_perio_1.tif |
Image of periosteum used for blinded scoring |
22324sp2_perio_2(1).tif |
Blinded image used in re-scoring set (for images differing by > 1 pt) |
22454sp2_perio_1(1).tif |
Blinded image used in re-scoring set (for images differing by > 1 pt) |
22332sp2_perio_1(1).tif |
Blinded image used in re-scoring set (for images differing by > 1 pt) |
22326sp2_perio_1(1).tif |
Blinded image used in re-scoring set (for images differing by > 1 pt) |
22435sp1_perio_1(1).tif |
Blinded image used in re-scoring set (for images differing by > 1 pt) |
22338sp1_perio_1(1).tif |
Blinded image used in re-scoring set (for images differing by > 1 pt) |
Raw_images_used_in_figures_251123.zip
- Description: Raw images of peripheral nerve and periosteum used to produce figures.
| File name | Description |
|---|---|
22455sp1_NormalNerve2a_40x.tif |
H&E image of a normal nerve used in Figure 1 |
22455sp2_Severe2a_40x.tif |
H&E image of a severe neuroma-like lesion used in Figure 1 |
22454sp1_NormalNerve1_trichrome_40x.tif |
Trichrome image of a normal nerve used in Figure 1 |
22324sp2_NerveProlif1_trichrome_40x.tif |
Trichrome image of a severe neuroma-like lesion used in Figure 1 |
22372sp1_perio_1.tif |
H&E image of a normal periosteum used in Figure 2 |
22422sp1b_perio_1.tif |
H&E image of a thickened periosteum nerve used in Figure 2 |
SheepBigSet_02Aug24_SS_22332sp2_s2_20x_CMY_Merge |
Composite immunostained image of normal nerve used in Figure 3 |
SheepBigSet_02Aug24_SS_22332sp2_s2_20x_CMY_NFH594_Magenta |
Immunostained image (targeting NFH) of normal nerve used in Figure 3 |
SheepBigSet_02Aug24_SS_22332sp2_s2_20x_CMY_S100488_Cyan |
Immunostained image (targeting s100) of normal nerve used in Figure 3 |
SheepBigSet_02Aug24_SS_22324sp2_s1_20x_CMY_Merge.tif |
Composite immunostained image of abnormal nerve used in Figure 4 |
SheepBigSet_02Aug24_SS_22324sp2_s1_20x_CMY_NFH594_Magenta.tif |
Immunostained image (targeting NFH) of abnormal nerve used in Figure 4 |
SheepBigSet_02Aug24_SS_22324sp2_s1_20x_CMY_S100488_Cyan.tif |
Immunostained image (targeting s100) of abnormal nerve used in Figure 4 |
22326sp2_nerve_20x2.tif |
H&E image of a normal nerve (fibrosis score 0) used in Figure S1 |
22338sp2_nerve_20x1.tif |
H&E image of a nerve that received a score of 1 for fibrosis used in Figure S1 |
22422sp1a_nerve_20x1.tif |
H&E image of a nerve that received a score of 2 for fibrosis used in Figure S1 |
22423sp1_nerve_20x1.tif |
H&E image of a nerve that received a score of 3 for fibrosis used in Figure S1 |
22455sp2_nerve_20x2.tif |
H&E image of a nerve that received a score of 4 for fibrosis used in Figure S1 |
22332sp1_nerve_20x1.tif |
H&E image of a normal nerve (axon sprouting score 0) used in Figure S2 |
22356sp2_nerve_20x1.tif |
H&E image of a nerve that received a score of 1 for axon sprouting used in Figure S2 |
22369sp1a_nerve_20x1.tif |
H&E image of a nerve that received a score of 2 for axon sprouting used in Figure S2 |
22435sp1_nerve_20x1.tif |
H&E image of a nerve that received a score of 3 for axon sprouting used in Figure S2 |
22503sp1_nerve_20x2.tif |
H&E image of a nerve that received a score of 4 for axon sprouting used in Figure S2 |
22355sp1_perio_1.tif |
H&E image of normal periosteum (periosteal thickening score 0) used in Figure S3 |
22372sp1_perio_1.tif |
H&E image of periosteum that received a score of 1 for periosteal thickening used in Figure S3 |
22434sp2_perio_1.tif |
H&E image of periosteum that received a score of 2 for periosteal thickening used in Figure S3 |
22435sp1_perio_1.tif |
H&E image of periosteum that received a score of 3 for periosteal thickening used in Figure S3 |
22454sp2_perio_3.tif |
H&E image of periosteum that received a score of 4 for periosteal thickening used in Figure S3 |
22324sp2_perio_2.tif |
H&E image of normal periosteum (periosteal axon proliferation score 0) used in Figure S4 |
22356sp1_perio_1.tif |
H&E image of periosteum that received a score of 1 for periosteal axon proliferation used in Figure S4 |
22423sp1_perio_1.tif |
H&E image of periosteum that received a score of 2 for periosteal axon proliferation used in Figure S4 |
22455sp2_perio_1.tif |
H&E image of periosteum that received a score of 3 for periosteal axon proliferation used in Figure S4 |
22502sp1_perio_2.tif |
H&E image of periosteum that received a score of 4 for periosteal axon proliferation used in Figure S4 |
22332sp2_s2_20x_CGRP647_NFH594_S100488_RGB_Merge.tif |
Composite immunostained image of normal nerve used in Figure S5 |
22332sp2_s2_20x_CGRP647_RGB_CGRP_BLUE.tif |
Immunostained image (targeting CGRP) of normal nerve used in Figure S5 |
22332sp2_s2_20x_NFH594_RGB.tif |
Immunostained image (targeting NFH) of normal nerve used in Figure S5 |
22332sp2_s2_20x_S100488_RGB.tif |
Immunostained image (targeting s100) of normal nerve used in Figure S5 |
22324sp2_s1_20x_CGRP647_NFH594_S100488_RGB_Merge.tif |
Composite immunostained image of abnormal nerve used in Figure S6 |
22324sp2_S1_20x_CGRP647_RGB_CGRP_BLUE.tif |
Immunostained image (targeting CGRP) of abnormal nerve used in Figure S6 |
22324sp2_S1_20x_NFH594_RGB.tif |
Immunostained image (targeting NFH) of abnormal nerve used in Figure S6 |
22324sp2_S1_20x_S100488_RGB.tif |
Immunostained image (targeting s100) of abnormal nerve used in Figure S6 |
