Retrospective analysis of post-mortem findings in Thoroughbreds aged from birth to 18 months presented to a UK pathology laboratory.

Enhanced understanding of reasons for, and timings of, mortality in Thoroughbreds prior to entering race training is warranted to provide insight into this population's health status. The aims of this study were to describe pathologies diagnosed at post-mortem (PM) examination in Thoroughbreds aged from birth to 18 months and investigate associations between age and pathology. Reports from a pathology laboratory in Newmarket, UK, were used to identify eligible cases examined between January 2006 and December 2020. Reported pathologies were extracted and categorised where appropriate. Comorbidities and pathogens identified were reported where available. Associations between age and selected pathologies were assessed using logistic regression. Of 144 eligible Thoroughbreds presented for PM, 137 had an available report and pathologist's diagnosis. Congenital defects were most commonly reported (20%; n = 28/137; 95%CI 15-29), 69% of which (n = 19/28; 95%CI 49-82) were conformational manifestations of developmental orthopedic disease (DOD). Pneumonia was an important pathology (14%; n = 20/137; 95%CI 36-53) during the pre-weaning period, where Rhodococcus equi was identified in 50% (n = 10/20; 95% CI 29-70) of cases. Odds of congenital defects (OR 56.6; 95%CI 7.0-460.0; P < 0.001) were significantly greater in horses aged 0-2 days compared to 4-18 months at PM. Odds of pneumonia (OR 4.3; 95%CI 1.1-1.7; P = 0.04) were significantly greater in horses ages 1-4 months compared to 0-2 days at PM. This study shows that conformational manifestations of DOD are an important contributor to perinatal mortality, and that pathologies reported at PM vary with age in young Thoroughbreds.

Association of neutrophil morphology with bacterial isolates in equine tracheal wash samples.

BACKGROUND: Tracheal wash sample neutrophilia is common in lower airway inflammation of various causes; however, relevance of cytomorphological features to culture of bacterial pathogens has not been established. OBJECTIVES: To investigate whether the presence of nondegenerate or degenerate neutrophils in tracheal washes is associated with culture of bacteria and, if so, whether this is influenced by age or temporal factors. STUDY DESIGN: Cross-sectional study. METHODS: Tracheal wash samples submitted to Rossdales LLP from 1/1/2013 to 31/7/2015 were evaluated using set criteria. Neutrophilia and degenerate neutrophilia (graded ≥2/4 on Rossdales cytological scale [0-4]) were analysed in relation to bacterial isolates considered potentially pathogenic in respiratory disease. Statistical analyses included multivariable logistic regression to identify associations between two separate outcomes: 1) the presence of neutrophilia compared with no neutrophilia and 2) the presence of degenerate neutrophilia compared with nondegenerate neutrophilia and four independent variables. RESULTS: Sufficient data for inclusion in the multivariable model for nondegenerate neutrophilia were available from 1100 horses. Culture of potentially pathogenic bacteria was associated with increased odds of degenerate neutrophilia compared with samples with negative culture (OR 4.5, 95% CI 3.1, 6.4, P-value<0.001). Horses over 9 years old had lower odds of having degenerate neutrophilia than those aged 1-3 years (OR 0.6, 95% CI 0.4, 0.9, P-value<0.02). In the spring/summer, horses had reduced odds of a degenerate neutrophilia compared with winter (OR 0.4, 95% CI 0.3, 0.7, P-value <0.001). MAIN LIMITATIONS: The study relied on routine laboratory submissions, with no control over sample collection or submitted clinical history. CONCLUSIONS: Cytological evaluation of tracheal washes should include cytomorphological features of the neutrophil response. The presence of degenerate neutrophils, especially in young horses, indicates added value of culture and sensitivity for antimicrobial therapeutics. The absence of degenerate changes, in combination with clinical factors, can help support diagnosis of a nonseptic cause of airway neutrophilia.

Serum bile acid concentrations, histopathological features, and short-, and long-term survival in horses with hepatic disease.

BACKGROUND: Serum bile acid concentrations (SBA) and a histopathological biopsy score [Equine Vet J 35 (2003) 534] are used prognostically in equine hepatic disease. HYPOTHESIS: Histopathologic features and scores, but not SBA, differ between survivors and nonsurvivors and correlate with histopathologic evidence of hepatic inflammation and fibrosis. ANIMALS: Retrospective study. Records (1999-2011) of horses with hepatic disease diagnosed by biopsy and with concurrent measurements of SBA. METHODS: Retrospective cohort study. Biopsies were examined for inflammatory cell infiltration including type and distribution, fibrosis, irreversible cytopathology affecting hepatocytes, hemosiderin, or other pigment deposition and bile duct proliferation. SBA, histopathological findings and a histological score [Equine Vet J 35 (2003) 534] were compared between short- (survival to discharge) and long-term (>6 months) survivors and correlations between SBA and histopathological findings investigated. RESULTS: Of 81 cases 90% survived short-term and 83% long-term. Short-term and long-term nonsurvival were associated with SBA (P = .009; P = .006), overall (P = .001; P = .002) and parenchymal (short-term only; P = .01) inflammation, portal and bridging fibrosis (all P < .001), apoptosis or single cell necrosis (P < .001; P = .008), hemosiderin deposition in hepatocytes (P = .011; P = .028), biliary (both P < .001), vascular (P = .003; P = .045) and endothelial (P < .001; P = .02) hyperplasia, nucleic changes (P = .004; P < .001) and the histopathological score (both P < .001). SBA were significantly and positively correlated with overall (P = .001), parenchymal (P < .001) and portal (P = .004) inflammation and portal (P = .036) and bridging (P = .002) fibrosis. CONCLUSIONS AND CLINICAL IMPORTANCE: SBA, histopathological findings and scores differ between survivors and nonsurvivors. SBA concentrations are associated with inflammation and fibrosis suggesting interference with hepatic function. A histopathological score >2 and, less so, SBA >20 µmol/L are specific but not sensitive indicators of nonsurvival.

Estrogen Receptor and Signal Transducer and Activator of Transcription 3 Expression in Equine Mammary Tumors.

Equine mammary tumors are uncommon, and relatively sparse histopathologic and molecular data exist. The present study describes the histopathologic features of 7 such tumors, which exhibited infiltrative growth, intermediate to high mitotic rates, and focally extensive necrosis. The tumors exhibited variably strong staining for vimentin and cytokeratin 14, as well as frequently weak cytoplasmic staining for pan-cytokeratin. E-cadherin expression was strong. Interestingly, a subgroup of the tumors exhibited strong nuclear staining for estrogen receptor α. Three of 7 tumors exhibited nuclear expression of the transcription factor STAT3, suggesting that STAT3 was transcriptionally active. Rare to absent nuclear STAT3 expression was observed in carcinomas exhibiting moderate to intense staining for cytokeratin 14. This investigation confirms previous investigators' assertions that equine mammary tumors have a malignant phenotype. A subset of the equine mammary tumors exhibited estrogen receptor α expression, suggesting that these tumors may potentially have similar molecular characteristics to their feline and canine counterparts.

Necrosis of the femoral condyles in a four-week-old foal: clinical, imaging and histopathological features.

A 4-week-old Thoroughbred filly foal with a history of sepsis was evaluated for right hindlimb lameness. Bilateral femoropatellar and femorotibial joint effusions were detected. Ultrasonography and radiography of the right stifle revealed signs of joint collapse and periarticular swelling. Computed tomography revealed abnormalities in the bone density of the medial femoral condyle of the right hindlimb and lateral femoral condyle of the left hindlimb. Euthanasia was recommended based on the severity of the lesions. Gross and microscopic examinations revealed extensive separation of the articular-epiphyseal cartilage complex from the subchondral bone of the femoral condyles. The histological features suggest an ischaemic aetiology; comparisons are made with lesions of osteochondrosis and avascular necrosis of bone.

A racing start in life? The hurdles of equine feto-placental pathology.

Significant progress has been made in understanding and monitoring the causes of equine abortion over past decades. However, not all in utero pathology results in abortion. It has long been recognised that some in utero pathology, such as twinning or chronic placentitis, can result in the birth of live but growth-retarded foals and there is historical evidence that birth weight may influence future athletic performance. Clinical experience (e.g. from twins) and experimental studies (pony-Thoroughbred embryo transfer) have highlighted the importance of reduced functional placental area in limiting growth in utero in horses. Many other nonfatal in utero pathologies (e.g. umbilical cord-related circulatory compromise) can potentially affect either placental function or other organ systems. Their influence on the short- and long-term health of the foal and its future athletic performance is in many cases poorly documented or understood. This review summarises the main causes of in utero pathology and reflects on how these may potentially affect the foal if born alive, highlighting the need for long-term studies on this important subject.

Salmonella enterica serovar typhimurium trxA mutants are protective against virulent challenge and induce less inflammation than the live-attenuated vaccine strain SL3261.

In Salmonella enterica serovar Typhimurium, trxA encodes thioredoxin 1, a small, soluble protein with disulfide reductase activity, which catalyzes thiol disulfide redox reactions in a variety of substrate proteins. Thioredoxins are involved as antioxidants in defense against oxidative stresses, such as exposure to hydrogen peroxide and hydroxyl radicals. We have made a defined, complete deletion of trxA in the mouse-virulent S. Typhimurium strain SL1344 (SL1344 trxA), replacing the gene with a kanamycin resistance gene cassette. SL1344 trxA was attenuated for virulence in BALB/c mice by the oral and intravenous routes and when used in immunization experiments provided protection against challenge with the virulent parent strain. SL1344 trxA induced less inflammation in murine spleens and livers than SL3261, the aroA mutant, live attenuated vaccine strain. The reduced splenomegaly observed following infection with SL1344 trxA was partially attributed to a reduction in the number of both CD4(+) and CD8(+) T cells and B lymphocytes in the spleen and reduced infiltration by CD11b(+) cells into the spleen compared with spleens from mice infected with SL3261. This less severe pathological response indicates that a trxA mutation might be used to reduce reactogenicity of live attenuated vaccine strains. We tested this by deleting trxA in SL3261. SL3261 trxA was also less inflammatory than SL3261 but was slightly less effective as a vaccine strain than either the SL3261 parent strain or SL1344 trxA.

Pathology of Mycobacterium bovis infection in wild meerkats (Suricata suricatta).

Pathological lesions associated with Mycobacterium bovis infection (bovine tuberculosis; bTB) in free-living meerkats (Suricata suricatta) in the Kalahari Desert of South Africa are described. The pathology of bTB in meerkats was determined through detailed post-mortem examinations of 57 animals (52 meerkats showing clinical signs of bTB, and five not showing signs of disease). Lymph nodes and tissue lesions thought to be associated with bTB were cultured for mycobacteria. All 52 bTB-infected meerkats showed gross or microscopical granulomatous lesions, but M. bovis was cultured from only 42% (22/52) of these animals. The majority (96%, 50/52) of diseased meerkats had lesions in multiple sites, the pattern of which suggested haematogenous spread of M. bovis infection in this species. The histological characteristics of the tuberculous lesions, together with the gross pathology and the wide range of body systems affected, indicate that infection in meerkats is acquired principally via the respiratory and oral routes, whereas excretion is most likely via the respiratory tract and suppurating skin wounds. Urine and faeces appear to be unlikely sources of infection. The findings of this study provide information on the transmission, pathogenesis and epidemiology of bTB in meerkats that is likely to be relevant to the understanding of M. bovis infection in other social mammal species such as the European badger (Meles meles).

An investigation into the suitability of a commercial real-time PCR assay to screen for Taylorella equigenitalis in routine prebreeding equine genital swabs.

REASONS FOR PERFORMING STUDY: Standard bacteriological methods for identifying Taylorella equigenitalis in cervical smears are time consuming. Therefore, a more rapid real-time PCR assay was evaluated for its suitability in screening swabs. OBJECTIVE: To compare the results of a commercially available real-time PCR assay with routine microbiological culture for the identification of T. equigenitalis, the causative organism of contagious equine metritis, in equine genital swab samples, under 'field trial' conditions. MATERIALS AND METHODS: Routine prebreeding genital swabs (n=2072) collected from Thoroughbred mares and stallions during 2009 were examined together with stored T. equigenitalis positive material. Swabs were cultured for T. equigenitalis using standard microbiological techniques. Bacterial lysates were isolated from the swabs and examined for the presence of a 16S DNA fragment of T. equigenitalis, using a commercial multiplex real-time PCR assay system. RESULTS: There was complete concordance between positive and negative results obtained by the 2 methods. Real-time PCR also detected T. equigenitalis DNA from swabs that were negative using standard microbiological culture after 6 months' storage at +4 degrees C but from which T. equigenitalis had been isolated following collection. The sensitivities of real-time PCR and bacterial culture were both 10(-3) (equivalent to 3 colony-forming units). CONCLUSION AND CLINICAL RELEVANCE: Routine bacterial culture of T. equigenitalis requires an incubation period of not less than 7 days before a conclusive negative result can be obtained, whereas bacterial extraction and real-time PCR assay can be completed in less than 6 h. The commercially-available PCR assay tested provided a rapid and reliable method for the identification of T. equigenitalis from equine genital swabs and could be usefully employed for the screening of mares and stallions for preseason Horserace Betting Levy Board (HBLB) Code of Practice and in other situations such as for bloodstock sales screening requirements, overcoming the current delays imposed by bacterial culture requirements. Its use could be quality assured by the existing HBLB biannual testing scheme for designated laboratories.

Repopulation of oligodendrocyte progenitor cell-depleted tissue in a model of chronic demyelination.

Some, but not all, chronically demyelinated multiple sclerosis (MS) lesions are depleted of oligodendrocyte progenitor cells (OPCs) suggesting that OPCs are destroyed during the process of demyelination and some factor impedes OPC repopulation of the depleted tissue. The chronically demyelinated axons in MS lie in an astrocytic environment and it has been proposed that this might impede entry of OPCs into such regions. By depleting a short length of spinal cord of its OPCs using 40 Gy of X-irradiation in both normal rats and rats with progressive myelin loss accompanied by an astrocytosis (taiep rats), we investigated whether such changes affect the ability of OPCs to repopulate OPC-depleted tissue. In both taiep and normal rats, the rate of repopulation decreases with age, but no difference was detected in the rate at which OPCs repopulated normally myelinated and chronically demyelinated and astrocytosed tissue. This indicates that, if the astrocytic environment of the taiep central nervous system (CNS) is comparable to that found in MS lesions, then the presence of chronically demyelinated axons and astrocytosis in chronic MS lesions does not represent a barrier to repopulation of the tissue by OPCs. However, similar to the situation in the normal adult rodent CNS, the rate of repopulation by endogenous OPCs in aged taiep rats is very slow, approximately 0.2 mm per week.

Repopulation of oligodendrocyte progenitor cell depleted tissue in a model of chronic demyelination.

Some, but not all chronically demyelinated MS lesions are depleted of oligodendrocyte progenitor cells (OPCs) suggesting that OPCs are destroyed during the process of demyelination and some factor impedes OPC repopulation of the depleted tissue. The chronically demyelinated axons in MS lie in an astrocytic environment and it has been proposed that this might impede entry of OPCs into such regions. By depleting a short length of spinal cord of its OPCs using 40 Gy of X-irradiation in both normal rats and rats with progressive myelin loss accompanied by an astrocytosis (taiep rats), we investigated whether such changes affect the ability of OPCs to repopulate OPC-depleted tissue. In both taiep and normal rats, the rate of repopulation decreases with age, but no difference was detected in the rate at which OPCs repopulated normally myelinated and chronically demyelinated and astrocytosed tissue. This indicates that, if the astrocytic environment of the taiep CNS is comparable to that found in MS lesions, then the presence of chronically demyelinated axons and astrocytosis in chronic MS lesions does not represent a barrier to repopulation of the tissue by OPCs. However, similar to the situation in the normal adult rodent CNS, the rate of repopulation by endogenous OPCs in aged taiep rats is very slow, approximately 0.2 mm per week.

Inflammation stimulates remyelination in areas of chronic demyelination.

A major challenge in multiple sclerosis research is to understand the cause or causes of remyelination failure and to devise ways of ameliorating its consequences. This requires appropriate experimental models. Although there are many models of acute demyelination, at present there are few suitable models of chronic demyelination. The taiep rat is a myelin mutant that shows progressive myelin loss and, by 1 year of age, its CNS tissue has many features of chronic areas of demyelination in multiple sclerosis: chronically demyelinated axons present in an astrocytic environment in the absence of acute inflammation. Using the taiep rat and a combination of X-irradiation and cell transplantation, it has been possible to address a number of questions concerning remyelination failure in chronic multiple sclerosis lesions, such as whether chronically demyelinated axons have undergone changes that render them refractory to remyelination and why remyelination is absent when oligodendrocyte progenitor cells (OPCs) are present. Our experiments show that (i) transplanted OPCs will not populate OPC-containing areas of chronic demyelination; (ii) myelination competent OPCs can repopulate OPC-depleted chronically demyelinated astrocytosed tissue, but this repopulation does not result in remyelination--closely resembling the situation found in some multiple sclerosis plaques; and (iii) the induction of acute inflammation in this non-remyelinating situation results in remyelination. Thus, we can conclude that axonal changes induced by chronic demyelination are unlikely to contribute to remyelination failure in multiple sclerosis. Rather, remyelination fails either because OPCs fail to repopulate areas of demyelination or because if OPCs are present they are unable to generate remyelinating oligodendrocytes owing to the presence of inhibitory factors and/or a lack of the stimuli required to activate these cells to generate remyelinating oligodendrocytes. This non-remyelinating situation can be transformed to a remyelinating one by the induction of acute inflammation.