Effects of routine treatment with nonsteroidal anti-inflammatory drugs at calving and when lame on the future probability of lameness and culling in dairy cows: A randomized controlled trial.

Claw horn lesions (CHL) are reported as the most common cause of lameness in intensive dairy systems. Despite their prevalence, the underlying pathological mechanisms and preventive strategies for CHL remain poorly understood. Recent advances have pointed to the role of inflammation in disease aetiopathogenesis. Moderating inflammation from first calving may lead to long-term benefits and a viable intervention for treating and preventing disease. We conducted a 34-mo randomized controlled trial to investigate the effects of routine treatment with the nonsteroidal anti-inflammatory drug ketoprofen at calving and during treatment for lameness, on the future probability of lameness and culling, caused by exposure to normal farm conditions. A cohort of dairy heifers were recruited from a single, commercial dairy herd between January 8, 2018, and June 22, 2020, and randomly allocated to one of 4 treatment groups before first calving. The lactating herd was lameness scored every 2 wk on a 0 to 3 scale, to identify animals that became lame (single score ≥2a) and hence required treatment. Animals in group 1 received a therapeutic trim and a hoof block on the sound claw (if deemed necessary) every time they were treated for lameness. Animals in group 2 received the same treatment as group 1 with the addition of a 3-d course of ketoprofen (single dose daily) every time they were treated for lameness. Animals in group 3 received the same treatment as group 2 with the addition of a 3-d course of ketoprofen (single dose daily) starting 24 to 36 h after each calving. Animals in group 4 received a 3-d course of ketoprofen (single dose daily) every time they were identified with lameness. No therapeutic trim was administered to this group, unless they were identified as severely lame (a single score ≥3a). Animals were followed for the duration of the study (ending October 23, 2020). Probability of lameness was assessed by a lameness outcome score collected every 14 d. Data on culling was extracted from farm records. One hundred thirty-two animals were recruited to each group, with data from 438 animals included in the final analysis (111 in group 1, 117 in group 2, 100 in group 3, and 110 in group 4). Mixed effect logistic regression models were used to evaluate the effect of treatment group on the ongoing probability of lameness. Compared with the control group (group 1), animals in group 3 were less likely to become lame (odds ratio: 0.66) and severely lame (odds ratio: 0.28). A Cox proportional hazards survival model was used to investigate the effect of treatment group on time to culling. Compared with group 1, animals in groups 2 and 3 were at reduced risk of culling (hazard ratios: 0.55 and 0.56, respectively). The lameness effect size we identified was large and indicated that treating a cohort of animals with the group 3 protocol, would lead to an absolute reduction in population lameness prevalence of approximately 10% and severe lameness prevalence of 3%, compared with animals treated in accordance with conventional best practice (group 1).

Interobserver agreement of digital dermatitis M-scores for photographs of the hind feet of standing dairy cattle.

Digital dermatitis (DD) is the leading infectious cause of lameness in dairy cattle, and it affects their welfare and productivity worldwide. At the herd level, DD is often assessed while cows are standing in a milking parlor, and lesions are most commonly evaluated using the M-score. The objective of this study was to examine the interobserver agreement for M-scores of the feet of standing cattle, based on digital color photographs of dairy cattle hind feet. A total of 88 photographs and written descriptors of the M-score were sent to 11 scorers working at 10 different institutions in 5 countries. The scorers received no formal training immediately before scoring the photographs; however, all regularly used the M-score to score DD. The answers for 36 photographs were excluded from the analysis because the photograph either had more than 1 M-stage as mode or not all scorers assigned an M-score to it. The M-scores of the 11 scorers from 52 photographs were available for analysis. Interobserver agreement was tested using Gwet's agreement coefficient (AC1) and the mode was assumed correct. Overall, moderate agreement emerged for the M-score (AC1 = 0.48). For the individual M-stages, almost perfect agreement existed for M0 (AC1 = 0.99), M1 (AC1 = 0.92), and M3 (AC1 = 0.82), and substantial agreement for M2 (AC1 = 0.61), M4 (AC1 = 0.65), and M4.1 (AC1 = 0.71). This outcome indicates the degree of individual variation in M-scoring in this context by unstandardized, experienced European observers, particularly for the M2, M4, and M4.1 stages. Standardized training is likely to improve the consistency of M-scoring and thus the generalizability of future DD research results on this important endemic disease.

Use of novel sensors combining local positioning and acceleration to measure feeding behavior differences associated with lameness in dairy cattle.

Time constraints for dairy farmers are an important factor contributing to the under-detection of lameness, resulting in delayed or missed treatment of lame cows within many commercial dairy herds. Hence, a need exists for flexible and affordable cow-based sensor systems capable of monitoring behaviors such as time spent feeding, which may be affected by the onset of lameness. In this study a novel neck-mounted mobile sensor system that combines local positioning and activity (acceleration) was tested and validated on a commercial UK dairy farm. Position and activity data were collected over 5 consecutive days for 19 high-yield dairy cows (10 lame, 9 nonlame) that formed a subset of a larger (120 cow) management group housed in a freestall barn. A decision tree algorithm that included sensor-recorded position and accelerometer data was developed to classify a cow as doing 1 of 3 categories of behavior: (1) feeding, (2) not feeding, and (3) out of pen for milking. For each classified behavior the mean number of bouts, the mean bout duration, and the mean total duration across all bouts was determined on a daily basis, and also separately for the time periods in between milking (morning = 0630-1300 h; afternoon = 1430-2100 h; night = 2230-0500 h). A comparative analysis of the classified cow behaviors was undertaken using a Welch t-test with Benjamini-Hochberg post-hoc correction under the null hypothesis of no differences in the number or duration of behavioral bouts between the 2 test groups of lame and nonlame cows. Analysis showed that mean total daily feeding duration was significantly lower for lame cows compared with non-lame cows. Behavior was also affected by time of day with significantly lower mean total duration of feeding and higher total duration of nonfeeding in the afternoons for lame cows compared with nonlame cows. The results demonstrate how sensors that measure both position and acceleration are capable of detecting differences in feeding behavior that may be associated with lameness. Such behavioral differences could be used in the development of predictive algorithms for the prompt detection of lameness as part of a commercially viable automated behavioral monitoring system.

A prospective cohort study of digital cushion and corium thickness. Part 1: Associations with body condition, lesion incidence, and proximity to calving.

Claw horn disruption lesions (CHDL) are a major cause of lameness in dairy cattle and are likely a result of excessive forces being applied to the germinal epithelium that produces the claw horn. The digital cushion is a connective tissue structure, containing depots of adipose tissue, that sits beneath the distal phalanx and has been shown to be thicker in fatter cows. Body condition score (BCS) loss is a risk factor for CHDL, and one possible explanation is that fat is mobilized from the digital cushion during negative energy balance, causing the digital cushion to thin and lose force-dissipating capacity, leading to disruption of claw horn growth. This prospective cohort study investigated the association between measures of body fat and sole soft tissue (SST) thickness (a combined measure of the corium and digital cushion beneath the distal phalanx) in a longitudinal manner. The SST of 179 cows in 2 high-yielding dairy herds were measured at 5 assessment points between 8 wk before and 35 wk postcalving. The BCS, back fat thickness (BFT), and lesion incidence were recorded. Data were analyzed in a 4-level mixed effects regression model, with the outcome being SST thickness beneath the flexor tuberosity of the distal phalanx. Data from 827 assessment points were available for analysis. The overall mean of SST was 4.99 mm (standard deviation: 0.95). The SST was thickest 8 wk before calving (5.22 mm, standard deviation: 0.91) and thinnest 1 wk postcalving (4.68 mm, standard deviation: 0.87), suggesting an effect of calving on SST. The BFT was positively correlated with SST in the model with a small effect size (a 10 mm decrease in BFT corresponded with a 0.13 mm decrease in SST), yet the nadir of BFT was 11.0 mm at 9 to 17 wk postcalving (when SST was ∼4.95 mm), rather than occurring with the nadir of SST immediately after calving. The SST also varied with other variables [e.g., cows that developed a sole ulcer or severe sole hemorrhage during the study had thinner SST (-0.24 mm)], except when a sole ulcer was present, when it was thicker (+0.53 mm). Cows that developed lesions had a thinner digital cushion before the lesion occurrence, which became thickened with sole ulcer presence, perhaps representing inflammation. Furthermore, although BFT was correlated with SST over time, SST may also have been influenced by other factors such as integrity of the suspensory apparatus, which could have a major effect on CHDL. Measures of body fat likely contributed to having thin SST, but other factors including calving, herd, and lesion presence also had an effect.

Randomised positive control trial of NSAID and antimicrobial treatment for calf fever caused by pneumonia.

One hundred and fifty-four preweaning calves were followed between May and October 2015. Calves were fitted with continuous monitoring temperature probes (TempVerified FeverTag), programmed so a flashing light emitting diode (LED) light was triggered following six hours of a sustained ear canal temperature of ≥39.7°C. A total of 83 calves (61.9 per cent) developed undifferentiated fever, with a presumptive diagnosis of pneumonia through exclusion of other calf diseases. Once fever was detected, calves were randomly allocated to treatment groups. Calves in group 1 (NSAID) received 2 mg/kg flunixin meglumine (Allevinix, Merial) for three consecutive days and group 2 (antimicrobial) received 6 mg/kg gamithromycin (Zactran, Merial). If fever persisted for 72 hours after the initial treatment, calves were given further treatment (group 1 received antimicrobial and group 2 received NSAID). Calves in group 1 (NSAID) were five times more likely (P=0.002) to require a second treatment (the antimicrobial) after 72 hours to resolve the fever compared with the need to give group 2 (antimicrobial) calves a second treatment (NSAID). This demonstrates the importance of ongoing monitoring and follow-up of calves with respiratory disease. However, of calves with fever in group 1 (NSAID), 25.7 per cent showed resolution following NSAID-only treatment with no detrimental effect on the development of repeated fever or daily live weight gain. This suggests that NSAID alone may be a useful first-line treatment, provided adequate attention is given to ongoing monitoring to identify those cases that require additional antimicrobial treatment.

A prospective cohort study of digital cushion and corium thickness. Part 2: Does thinning of the digital cushion and corium lead to lameness and claw horn disruption lesions?

The aim of this study was to determine whether a decrease in thickness of the sole soft tissues (SST) beneath the flexor tuberosity of the distal phalanx (i.e., the digital cushion and corium) predisposed a claw to develop claw horn disruption lesions (CHDL) or a leg to lameness. Data were analyzed from a longitudinal study of 179 cows, which had been examined at 5 assessment points -8, +1, +9, +17, and +29 wk relative to their first, second, third, or fourth calving. At each assessment point, SST were measured using ultrasonography. Additional assessment point data included sole lesions and back fat thickness (BFT), and cows had been locomotion scored every 2 wk from calving. One hundred fifty-eight cows completed the study. Separate logistic regression survival analyses were constructed to assess the outcomes, either lameness on a leg or CHDL on a claw; combinations of lameness and lesions were tested as outcomes. Cow level variables tested included farm and lactation number. Variables were tested describing previous SST thickness, minimum previous SST thickness, BFT, and change in either variable between prior assessment points. Prior lesions/lameness strongly predicted repeat cases and the final models had the outcome first lesion or lameness on a claw or leg. In the reported lameness models, lameness was defined as a leg being recorded as lame twice within 3 consecutive scores, and in the reported lesion models, lesion was defined as the first presence of either a sole ulcer or a severe sole hemorrhage on a claw. Thin SST increased the likelihood of lesion occurrence; thin SST on the lateral claw predicted subsequent lameness on a leg. Thin BFT and thinning of BFT between previous assessment points increased the likelihood of future lesion occurrence. Thin SST and thinning of BFT had additional effects on the likelihood of lesion occurrence, suggesting that BFT and sole SST had independent effects on lesion occurrence. However, change in SST thickness between assessment points did not influence the likelihood of future lesions or lameness. This suggests that thin SST were not simply a result of depletion of body fat and challenges the theory that thinning of the digital cushion with body fat mobilization leads to CHDL. Other possible mechanisms by which SST become thin are discussed and could include changes in integrity of the suspensory apparatus with physiological events.

Randomised controlled trial to evaluate the effect of foot trimming before and after first calving on subsequent lameness episodes and productivity in dairy heifers.

The objective of this study was to assess both independent and combined effects of routine foot trimming of heifers at 3 weeks pre-calving and 100 days post calving on the first lactation lameness and lactation productivity. A total of 419 pre-calving dairy heifers were recruited from one heifer rearing operation over a 10-month period. Heifers were randomly allocated into one of four foot trimming regimens; pre-calving foot trim and post-calving lameness score (Group TL), pre-calving lameness score and post-calving foot trim (Group LT), pre-calving foot trim and post-calving foot trim (Group TT), and pre-calving lameness score and post-calving lameness score (Group LL, control group). All heifers were scored for lameness at 24 biweekly time points for 1 year following calving, and first lactation milk production data were collected. Following calving, 172/419 (41.1%) of heifers became lame during the study (period prevalence), with lameness prevalence at each time-point following calving ranging from 48/392 (12.2%) at 29-42 days post-calving to 4/379 (1.1%) between 295 and 383 days after calving. The effects of the four treatment groups were not significantly different from each other for overall lameness period prevalence, biweekly lameness point prevalence, time to first lameness event, type of foot lesion identified at dry off claw trimming, or the 4% fat corrected 305-day milk yield. However, increased odds lameness was significantly associated with a pre-calving trim alone (P = 0.044) compared to the reference group LL. The odds of heifer lameness were highest between 0 and 6 weeks post-partum, and heifer farm destination was significantly associated with lameness (OR 2.24), suggesting that even at high standard facilities, environment and management systems have more effect on heifer foot health than trimming.

Linking bone development on the caudal aspect of the distal phalanx with lameness during life.

Claw horn disruption lesions (CHDL; sole hemorrhage, sole ulcer, and white line disease) cause a large proportion of lameness in dairy cattle, yet their etiopathogenesis remains poorly understood. Untreated CHDL may be associated with damage to the internal anatomy of the foot, including to the caudal aspect of the distal phalanx upon which bone developments have been reported with age and with sole ulcers at slaughter. The primary aim of this study was to assess whether bone development was associated with poor locomotion and occurrence of CHDL during a cow's life. A retrospective cohort study imaged 282 hind claws from 72 Holstein-Friesian dairy cows culled from a research herd using X-ray micro-computed tomography (µ-CT; resolution: 0.11mm). Four measures of bone development were taken from the caudal aspect of each distal phalanx, in caudal, ventral, and dorsal directions, and combined within each claw. Cow-level variables were constructed to quantify the average bone development on all hind feet (BD-Ave) and bone development on the most severely affected claw (BD-Max). Weekly locomotion scores (1-5 scale) were available from first calving. The variables BD-Ave and BD-Max were used as outcomes in linear regression models; the explanatory variables included locomotion score during life, age, binary variables denoting lifetime occurrence of CHDL and of infectious causes of lameness, and other cow variables. Both BD-Max and BD-Ave increased with age, CHDL occurrence, and an increasing proportion of locomotion scores at which a cow was lame (score 4 or 5). The models estimated that BD-Max would be 9.8mm (SE 3.9) greater in cows that had been lame at >50% of scores within the 12mo before slaughter (compared with cows that had been assigned no lame scores during the same period), or 7.0mm (SE 2.2) greater if the cow had been treated for a CHDL during life (compared with cows that had not). Additionally, histology demonstrated that new bone development was osteoma, also termed "exostosis." Age explained much of the variation in bone development. The association between bone development and locomotion score during life is a novel finding, and bone development appears specific to CHDL. Bone development on the most severely affected foot was the best explained outcome and would seem most likely to influence locomotion score. To stop irreparable anatomical damage within the foot, early identification of CHDL and effective treatment could be critical.

Recovery of chronically lame dairy cows following treatment for claw horn lesions: a randomised controlled trial.

A positively controlled, randomised controlled trial (RCT) was undertaken to test recovery of cows with claw horn lesions resulting in lameness of greater than two weeks duration. Cows on seven commercial farms were mobility scored fortnightly and selected by lameness severity and chronicity. Study cows all received a therapeutic trim then random allocation of: no further treatment (trim only (TRM)), plastic shoe (TS) or plastic shoe and NSAID (TSN). Recovery was assessed by mobility score at 42 (±4) days post treatment by an observer blind to treatment group. Multivariable analysis showed no significant effect of treatment with an almost identical, low response rate to treatment across all groups (Percentage non-lame at outcome: TRM--15 per cent, TS--15 per cent, TSN--16 per cent). When compared with results of a similar RCT on acutely lame cows, where response rates to treatment were substantially higher, it can be concluded that any delay in treatment is likely to reduce the rate of recovery, suggesting early identification and treatment is key. Thirty-eight per cent of animals treated in this study were lame on the contralateral limb at outcome suggesting that both hindlimbs should be examined and a preventive or if necessary a therapeutic foot trim performed when lameness is identified particularly if the duration of lameness is unknown.

Evaluation of treatments for claw horn lesions in dairy cows in a randomized controlled trial.

Lameness is one of the most significant endemic disease problems facing the dairy industry. Claw horn lesions (principally sole hemorrhage, sole ulcer, and white line disease) are some of the most prevalent conditions. Despite the fact that thousands of animals are treated for these conditions every year, experimental evidence is limited on the most effective treatment protocols. A randomized, positively controlled clinical trial was conducted to test the recovery of newly lame cows with claw horn lesions. Animals on 5 farms were locomotion scored every 2wk. Cows were eligible for recruitment if they had 2 nonlame scores followed by a lame score and had a claw horn lesion on a single claw of a single foot. Following a therapeutic trim, enrolled cows were randomly allocated to 1 of 4 treatments: treatment 1-no further treatment (positive control; TRM), treatment 2-trim plus a block on the sound claw (TB), treatment 3-trim plus a 3-d course of the nonsteroidal anti-inflammatory drug (NSAID) ketoprofen (TN), treatment 4-trim plus a block plus ketoprofen (TBN). The primary outcome measure was locomotion score 35d after treatment, by an observer blind to treatment group. Descriptive statistics suggested that treatment groups were balanced at the time of enrollment, that is, randomization was successful. Based on a sound locomotion score (score 0) 35d after treatment, the number of cures was 11 of 45 (24.4%) for TRM, 14 of 39 (35.9%) for TB, 12 of 42 (28.6%) for TN, and 23 of 41 (56.1%) for TBN. The difference between TBN and TRM was significant. To test for confounding imbalances between treatment groups, logistic regression models were built with 2 outcomes, either sound (score 0) or nonlame (score 0 or 1) 35d after treatment. Compared with TRM, animals that received TBN were significantly more likely to cure to a sound outcome. Farm, treatment season, lesion diagnosis, limb affected, treatment operator, and stage of lactation were included in the final models. Our work suggests that lameness cure is maximized with NSAID treatment in addition to the common practices of therapeutic trimming and elevation of the diseased claw using a block when cows are newly and predominantly mildly lame.

Measuring the response to therapeutic foot trimming in dairy cows with fortnightly lameness scoring.

Lameness scoring (0-3) was carried out on four UK dairy farms during the housing period over three consecutive years (2010-2012). At the start of the study cows were matched by parity and stage of lactation and randomly allocated into a treatment (TX) and a control (CX) group. Cows were enrolled when two sound scores (0 or 1) were followed by a lame score (2). Farmers were immediately notified of score 3 cows, which were then excluded from the study, irrespective of whether they were in treatment or control groups. The animals in the TX group received treatment 3-48 h after being scored lame. Farmers remained blind to the treatment group. Throughout the study the participating farmers continued to identify and treat lame cows according to their usual approaches, this included treating animals in the CX or TX group if they so chose. The fortnightly lameness scoring and treatment of the TX group resulted in higher cure rates at each scoring session following treatment when compared with the CX group (P < 0.001). Two weeks after inclusion, 78% (SE ± 3.2) of TX cows were sound, compared with 66% (SE ± 3.1) of CX cows. At 18 weeks following initial recruitment this had fallen to 41% (SE ± 6.3) (TX) and 13% (SE ± 4.7) (CX). The percentage of total scores which were sound scores in the TX and CX groups following inclusion in the trial was 81% and 66.1%, respectively (P < 0.001). The main lesions found on treatment in the TX group were sole haemorrhage (41% of cases) and digital dermatitis (33%). Severe lesions (sole ulcers and toe necrosis) were only found in 6.6% of cases. In the treated CX animals the percentage of severe lesions was 14%.

A survey of the on-farm treatment of sole ulcer and white line disease in dairy cattle.

A telephone survey of UK dairy farmers was conducted to investigate current on-farm practice in the treatment of mild sole ulcer (SU)/sole bruising (SB), and white line disease (WLD), and the potential barriers associated with therapy. A total of 84 dairy farmers were questioned about the process of detecting and treating lame cows on their farm as well as about the specific treatments they applied. Farmers were also canvassed for their views on the efficacy of different potential treatments for mild SU/SB and WLD. In general, respondents discussed treatments for SU and WLD rather than specifically for mild SU/SB and WLD. Furthermore, when describing treatment methods, farmers rarely differentiated between SU and WLD. Trimming the affected claw with or without the additional use of orthopaedic blocks was the most commonly reported treatment method considered effective and practical by the majority of farmers. Antibiotics and/or analgesics were used by a small number of farmers, and some housed their most severely lame cows in straw pens. Lack of time, inadequate equipment and poor farm layout were identified by many survey respondents as barriers to the prompt and/or effective treatment of their lame animals.

A descriptive review of the peer and non-peer reviewed literature on the treatment and prevention of foot lameness in cattle published between 2000 and 2011.

The aim of this study was to collate and review the peer and non-peer reviewed English language literature on the treatment and prevention of foot lameness in cattle published since January 2000. The study aimed to identify deficits in knowledge and areas of disparity between what is recommended in the field by veterinarians, foot trimmers and advisors and what has been substantiated experimentally. Peer reviewed literature containing original work was gathered by searching three databases. Papers were categorised and reviewed if they contained material on treatment or prevention. Non-peer reviewed clinical materials were collated from a range of sources. The materials were reviewed and categorised based on whether they recommended a range of possible treatment and prevention strategies. The peer reviewed data base contained 591 papers, of which 286 contained information on treatment or prevention. The vast majority of papers (258) concerned prevention; only a small number covered treatment (31) and of these only three contained information on the treatment of sole ulcers or white line disease. The number of intervention studies and trials was low; most papers on prevention were observational. Generally, lesion specific outcomes were not described making the findings of these papers difficult to use clinically. The non-peer reviewed material contained 46 sources; they varied significantly in regard to the treatments they advocated with some texts directly contradicting each other. Some aspects of prevention recommended in these sources seemed poorly supported by findings from the research literature. Well designed intervention studies are required to address these deficits.

The effects of early treatment for hindlimb lameness in dairy cows on four commercial UK farms.

An 'early threshold' protocol for treating cows within 48 h of being detected lame in one or more hind limbs at fortnightly mobility scoring was tested on a randomly selected group of cattle on four commercial dairy farms. The outcomes of the early threshold treatment for first cases of lameness were compared with those of the farmers' conventional approach to treatment. The early threshold schedule resulted in a much shorter time to treatment than the conventional approach, for which the median time from the cow first being scored lame to treatment was 65 days. The early threshold group presented with less severe foot lesions and cattle were less likely to be selected for further treatments by the farmer than conventionally treated cows. Early threshold treatment reduced the prevalence of lameness 4 weeks after treatment, compared with controls. A clear effect of the early treatment on milk yield was not detected.

Evaluating an intervention to reduce lameness in dairy cattle.

Lameness in dairy cattle remains a significant welfare concern for the UK dairy industry. Farms were recruited into a 3-yr study evaluating novel intervention approaches designed to encourage farmers to implement husbandry changes targeted toward reducing lameness. All farms completing the study were visited at least annually and received either monitoring only (MO, n=72) or monitoring and additional support (MS, n = 117) from the research team. The additional support included traditional technical advice on farm-specific solutions, facilitation techniques to encourage farmer participation, and application of social marketing principles to promote implementation of change. Lameness prevalence was lower in the MO (27.0 ± 1.94 SEM) and MS (21.4 ± 1.28) farms at the final visit compared with the same MO (38.9 ± 2.06) and MS (33.3 ± 1.76) farms on the initial visit. After accounting for initial lameness, intervention group status, and year of visit within a multilevel model, we observed an interaction between year and provision of support, with the reduction in lameness over time being greater in the MS group compared with the MO group. Farms in the MS group made a greater number of changes to their husbandry practices over the duration of the project (8.2 ± 0.39) compared with those farms in the MO group (6.5 ± 0.54). Because the lameness prevalence was lower in the MS group than the MO group at the start of the study, the contribution of the additional support was difficult to define. Lameness can be reduced on UK dairy farms although further work is needed to identify the optimum approaches.

Haemosiderin deposition in Donkey (Equusasinus) liver: Comparison of liver histopathology with liver iron content.

Histopathological examination was carried out on post mortem samples of liver from 12 donkeys (Equus asinus), aged 21-57 years (4 females, 1 stallion, 7 geldings). Variable amounts of haemosiderin were present in Kupffer cells, portal macrophages and hepatocytes in all cases. In all cases there was infiltration of connective tissue around portal tracts by variable numbers of inflammatory cells (lymphocytes, plasma cells and macrophages) but obvious portal fibrosis was present in only four animals. Subjective assessment of overall haemosiderin staining (including extent and intensity) generally reflected biochemical measurements of liver iron content (measured by an inductively-coupled plasma method) as well as quantitative histochemical measurements (using an image analysis package and sections stained with Perl's Prussian blue stain). Accumulation of hepatic iron in old donkeys was not directly related to other pathological changes and may be an incidental finding.