Lubricin/proteoglycan 4 increases in both experimental and naturally occurring equine osteoarthritis.

OBJECTIVE: The goals of this study were (1) to quantify proteoglycan 4 (PRG4) gene expression; (2) to assess lubricin immunostaining; and (3) to measure synovial fluid lubricin concentrations in clinical and experimental models of equine carpal osteoarthritis (OA). DESIGN: Lubricin synovial fluid concentrations and cartilage and synovial membrane PRG4 expression were analyzed in research horses undergoing experimental OA induction (n = 8) and in equine clinical patients with carpal OA (n = 58). Lubricin concentrations were measured using a custom sandwich enzyme-linked immunosorbent assay, and PRG4 expression was quantified using qRT-PCR. Lubricin immunostaining was assessed in synovial membrane and osteochondral sections in the experimental model. RESULTS: Lubricin concentrations increased in synovial fluid following induction of OA, peaking at 21 days post-operatively in OA joints vs sham-operated controls (331 ± 69 µg/mL vs 110 ± 19 µg/mL, P = 0.001). Lubricin concentrations also increased in horses with naturally occurring OA as compared to control joints (152 ± 32 µg/mL vs 68 ± 4 µg/mL, P = 0.003). Synovial membrane PRG4 expression increased nearly 2-fold in naturally occurring OA (P = 0.003), whereas cartilage PRG4 expression decreased 2.5-fold (P = 0.025). Lubricin immunostaining was more pronounced in synovial membrane from OA joints as compared to controls, with intense lubricin localization to sites of cartilage damage. CONCLUSIONS: Although PRG4 gene expression decreases in OA cartilage, synovial membrane PRG4 expression, synovial fluid lubricin concentrations and lubricin immunostaining all increase in an equine OA model. Lubricin may be elevated to protect joints from post-traumatic OA.

Identification of cartilage injury using quantitative multiphoton microscopy.

OBJECTIVE: Cartilage injury can lead to post-traumatic osteoarthritis (PTOA). Immediate post-trauma cellular and structural changes are not widely understood. Furthermore, current cellular-resolution cartilage imaging techniques require sectioning of cartilage and/or use of dyes not suitable for patient imaging. In this study, we used multiphoton microscopy (MPM) data with FDA-approved sodium fluorescein to identify and evaluate the pattern of chondrocyte death after traumatic injury. METHOD: Mature equine distal metacarpal or metatarsal osteochondral blocks (OCBs) were injured by 30 MPa compressive loading delivered over 1 s. Injured and control sites were imaged unfixed and in situ 1 h post-injury with sodium fluorescein using rasterized z-scanning. MPM data was quantified in MATLAB, reconstructed in 3-D, and projected in 2-D to determine the damage pattern. RESULTS: MPM images (600 per sample) were reconstructed and analyzed for cell death. The overall distribution of cell death appeared to cluster into circular (n = 7) or elliptical (n = 4) patterns (p = 0.006). Dead cells were prevalent near cracks in the matrix, with only 26.3% (SE = 5.0%, p < 0.0001) of chondrocytes near cracks being viable. CONCLUSION: This study demonstrates the first application of MPM for evaluating cellular-scale cartilage injury in situ in live tissue, with clinical potential for detecting early cartilage damage. With this technique, we were able to uniquely observe two death patterns resulting from the same compressive loading, which may be related to local variability in matrix structure. These results also demonstrate proof-of-concept MPM diagnostic use in detecting subtle and early cartilage damage not detectable in any other way.

Clinical reappraisal of the composite international diagnostic interview version 3.3 in Qatar's National Mental Health Study.

OBJECTIVES: Lifetime DSM-5 diagnoses generated by the lay-administered Composite International Diagnostic Interview for DSM-5 (CIDI) in the World Mental Health Qatar (WMHQ) study were compared to diagnoses based on blinded clinician-administered reappraisal interviews. METHODS: Telephone follow-up interviews used the non-patient edition of the Structured Clinician Interview for DSM-5 (SCID) oversampling respondents who screened positive for five diagnoses in the CIDI: major depressive episode, mania/hypomania, panic disorder, generalized anxiety disorder, and obsessive-compulsive disorder. Concordance was also examined for a diagnoses of post-traumatic stress disorder based on a short-form versus full version of the PTSD Checklist for DSM-5 (PCL-5). RESULTS: Initial CIDI prevalence estimates differed significantly from the SCID for most diagnoses ( χ 1 2 ${\chi }_{1}^{2}$  = 6.6-31.4, p = 0.010 < 0.001), but recalibration reduced most of these differences and led to consistent increases in individual-level concordance (AU-ROC) from 0.53-0.76 to 0.67-0.81. Recalibration of the short-form PCL-5 removed an initially significant difference in PTSD prevalence with the full PCL-5 (from χ 1 2 ${\chi }_{1}^{2}$  = 610.5, p < 0.001 to χ 1 2 ${\chi }_{1}^{2}$  = 2.5, p = 0.110) while also increasing AU-ROC from 0.76 to 0.81. CONCLUSIONS: Recalibration resulted in valid diagnoses of common mental disorders in the Qatar National Mental Health Survey, but with inflated prevalence estimates for some disorders that need to be considered when interpreting results.

The risk of Listeria monocytogenes infection in beef cattle operations.

AIM: To determine the prevalence of Listeria monocytogenes and associated risk factors among beef operations (cow-calf and feedlot) in central and southern California. METHODS AND RESULTS: A repeated cross-sectional study where faecal and environmental samples were collected from 50 operations three times a year at different seasons was carried out. Samples were tested for presence of L. monocytogenes using a combination of enrichment and polymerase chain reaction tests. Data on putative risk factors were also collected. Listeria monocytogenes was detected in faecal samples from cows, calves and other animals on calf-cow operations at proportions of 3.1%, 3.75% and 2.5%, respectively. The organism was detected in 5.3% of cut-grass, 5.3% of soil, 14.3% of irrigation ditches, 3.1% of the ponds and 6.5% of water troughs samples. Listeria monocytogenes was less common in faecal (0.3%) and soil (0.75%) samples collected from feedlots. CONCLUSIONS: Listeria monocytogenes was present at a higher proportion among cow-calf operations than feedlots. There was no significant seasonal variation in the occurrence of this pathogen within the two types of operations. SIGNIFICANCE AND IMPACT OF THE STUDY: If risk mitigation strategies were implemented to reduce the public health risk these should focus in cow-calf operations.

Relationships between race earnings and horse age, sex, gait, track surface and number of race starts for Thoroughbred and Standardbred racehorses in North America.

REASONS FOR PERFORMING STUDY: There is no consensus on objective outcome measures that can be used to determine if a medical or surgical treatment affects race performance. OBJECTIVE: To determine the association between 2 commonly used outcome measures (total starts and total earnings) and age, sex, gait and race surface. METHODS: A cross-sectional study was performed using the race performance data for all Thoroughbred horses age 2, 3, 4 and 5 years racing in the United States, and Standardbred horses of the same ages racing in the United States and Canada during the year 2006. Median earnings and starts were determined for each combination of age, sex and track surface (for Thoroughbred) or gait (for Standardbred). The effect these variables had on starts on race earnings ($) was determined using linear regression. RESULTS: Race records for 68,649 Thoroughbreds and 25,830 Standardbreds were obtained. All independent variables (age, breed, sex, gait, track surface and total number of starts) had a significant impact on total earnings (P<0.0001). CONCLUSIONS: The data show considerable variation across age groups and track surfaces for Thoroughbreds and across age groups for Standardbreds. They also show that the decision to use earnings or starts as outcome measures could have a marked effect on reported success for a particular treatment. POTENTIAL RELEVANCE: Both earning and start data should be reported in studies evaluating outcome following surgery or other intervention. Considerations of age, breed, sex, track surface and gait should be included in the design of these studies.

Racing performance after combined prosthetic laryngoplasty and ipsilateral ventriculocordectomy or partial arytenoidectomy: 135 Thoroughbred racehorses competing at less than 2400 m (1997-2007).

REASONS FOR PERFORMING STUDY: The success of combined prosthetic laryngoplasty with ipsilateral ventriculocordectomy (LPVC) has not been compared to that of partial arytenoidectomy (PA) in a clinical population. HYPOTHESES: In Thoroughbred (TB) racehorses: (1) earnings after LPVC are unaffected by the severity of recurrent laryngeal neuropathy (RLN) (laryngeal grade III vs. grade IV); (2) LPVC and PA yield similar results in the treatment of grade III RLN; (3) performance outcome following PA is independent of diagnosis (RLN vs. unilateral arytenoid chondritis [UAC]); and (4) neither LPVC nor PA returns horses to the level of performance of controls. METHODS: Medical and racing records of 135 TB racehorses undergoing LPVC or PA for the treatment of grade III or IV RLN or UAC were reviewed. Racing records of age and sex matched controls were also reviewed. RESULTS: After LPVC, horses with grade III RLN performed better compared to those with grade IV RLN. Furthermore, horses treated for grade III RLN by LPVC showed post operative earnings comparable to controls. Rate of return to racing were similar for PA and LPVC, although LPVC resulted in higher post operative earnings. Performance after PA was similar regardless of diagnosis (UAC or RLN). Finally, neither LPVC when performed for grade IV RLN, nor PA performed for either diagnosis restored post operative earnings to control levels. CONCLUSIONS: Thoroughbred racehorses treated by LPVC for grade III RLN show significantly better post operative earnings compared to horses treated for grade IV disease. In grade III RLN, LPVC returns earning potential to control levels. PA and LPVC lead to similar success in terms of rate of return to racing, but PA leads to inferior earnings after surgery. POTENTIAL RELEVANCE: Laryngoplasty should be recommended for all TB racehorses with grade III RLN to maximise return to racing at a high level. This contradicts the common approach of waiting for complete paralysis.

Crib-biting in US horses: breed predispositions and owner perceptions of aetiology.

REASONS FOR PERFORMING STUDY: Crib-biting is an equine stereotype that may result in diseases such as colic. Certain breeds and management factors have been associated. OBJECTIVES: To determine: breed prevalence of crib-biting in US horses; the likelihood that one horse learns to crib-bite from another; and owner perceptions of causal factors. METHODS: An initial postal survey queried the number and breed of crib-biting horses and if a horse began after being exposed to a horse with this habit. In a follow-up survey, a volunteer subset of owners was asked the number of affected and nonaffected horses of each breed and the extent of conspecific contact. The likelihood of crib-biting given breed and extent of contact was quantified using odds ratio (OR) and significance of the association was assessed using the Chi-squared test. RESULTS: Overall prevalence was 4.4%. Thoroughbreds were the breed most affected (133%). Approximately half of owners believed environmental factors predominantly cause the condition (54.4%) and crib-biting is learned by observation (48.8%). However, only 1.0% of horses became affected after being exposed to a crib-biter. The majority (86%) of horses was turned out in the same pasture with other horses and extent of contact with conspecifics was not statistically related to risk. CONCLUSION: This is the first study to report breed prevalence for crib-biting in US horses. Thoroughbreds were the breed more likely to be affected. More owners believed either environmental conditions were a predominant cause or a combination of genetic and environmental factors contributes to the behaviour. Only a small number of horses reportedly began to crib-bite after being exposed to an affected individual, but approximately half of owners considered it to be a learned behaviour; most owners did not isolate affected horses. POTENTIAL RELEVANCE: Genetic predisposition, not just intensive management conditions and surroundings, may be a factor in the high crib-biting prevalence in some breeds, and warrants further investigation. Little evidence exists to suggest horses learn the behaviour from other horses, and isolation may cause unnecessary stress.

Risk assessment for Escherichia coli O157:H7 in marketed unpasteurized milk in selected East African countries.

We carried out a study to assess the risk associated with the presence of Shiga toxigenic Escherichia coli (STEC) in informally marketed unpasteurized milk in urban East Africa. Data for the risk models were obtained from on-going and recently completed studies in Kenya and Uganda. Inputs for the model were complemented with data from published literature in similar populations. A fault-tree scenario pathway and modular process risk model approach were used for exposure assessment. Hazard characterization was based on a socioeconomic study with dose-responses derived from the literature. We used a probabilistic approach with Monte Carlo simulation and inputs from farm and household surveys. The qualitative analysis suggested a low to moderate risk of infection from consuming milk and that the widespread consumer practice of boiling milk before consumption was an important risk mitigator. Quantitative analysis revealed that two to three symptomatic STEC infections could be expected for every 10,000 unpasteurized milk portions consumed, with a possible range of 0 to 22 symptomatic cases. Sensitivity analyses to assess the uncertainty and variability associated with the model revealed that the factor with the greatest influence on disease incidence was the prevalence of STEC in dairy cattle. Risk assessment is a potentially useful method for managing food safety in informal markets.

Ocular penetration of intravenously administered enrofloxacin in the horse.

REASON FOR PERFORMING STUDY: Information on antibiotic concentrations in the equine eye following systemic therapy is limited. Reports that Leptospira spp. are frequently present in the eyes of horses with recurrent uveitis, emphasises a need for studies on ocular concentrations of specific antibiotics. HYPOTHESES: 1) Enrofloxacin, administered i.v. at 7.5 mg/kg bwt q. 24 h, results in aqueous humour concentrations greater than the reported minimum inhibitory concentration (MIC) for Leptospira pomona. 2) Aqueous humour paracentesis sufficiently disrupts the blood-aqueous humour barrier (BAB) to cause an increase in aqueous humour protein and enrofloxacin concentrations. METHODS: Aqueous humour enrofloxacin and total protein concentrations were determined in 6 healthy, mature horses after i.v. administration of enrofloxacin. Paracentesis was performed on the left eye on Days 3 and 4, 1 h following enrofloxacin administration, to determine enrofloxacin concentrations in healthy eyes and in eyes with mechanical disruption of the BAB. Paracentesis was also performed on the right eye 23 h after enrofloxacin administration. Blood samples were collected from the horses at identical times to determine enrofloxacin aqueous humour:plasma ratios. RESULTS: Mean +/- s.d. enrofloxacin concentration in the aqueous humour 1 h post administration on Day 3 was 0.32 +/- 0.10 mg/l (range 0.18-0.47); and aqueous humour enrofloxacin, total protein and aqueous humour:plasma enrofloxacin ratios were higher on Day 4 than Day 3. CONCLUSIONS AND POTENTIAL RELEVANCE: Following disruption of the BAB, enrofloxacin concentrations were above the reported MIC for Leptospira pomona.

Intra-articular stabilisation of the equine cricoarytenoid joint.

REASONS FOR PERFORMING STUDY: The success of laryngoplasty is limited by abduction loss in the early post operative period. OBJECTIVE: To determine the efficacy of polymethylmethacrylate (PMMA) in stabilising the cricoarytenoid joint (CAJ) and reducing the force on the laryngoplasty suture. HYPOTHESIS: Injection into the cricoarytenoid joint resists the forces produced by physiological laryngeal air flows and pressures thereby reducing the force experienced by the laryngoplasty suture. METHODS: Ten cadaver larynges were collected at necropsy and PMMA was injected into one CAJ at selected random. Each larynx was subjected to physiological conditions with with constant (static) or cycling (dynamic) flow. The specimens were tested sequentially in each of 4 conditions: 1) bilateral full abduction (Control 1); 2) transection of the suture on the side without PMMA; 3) bilateral abduction achieved by replacing the suture (Control 2); and 4) cutting the suture on the PMMA side. Tracheal pressure and flow and pressure in the flow chamber were recorded using pressure and flow transducers. The strain experienced by each suture during bilateral abduction (Controls 1 and 2) was measured. Statistical comparison of the 4 conditions was performed using a mixed effect model with Tukey's post hoc test for multiple comparisons. The strain gauge data were analysed by paired comparison of the regression slopes. RESULTS: In the static and dynamic states, tracheal pressure increased and tracheal flow decreased when the suture on the non-cement side was cut (P < 0.05). There was no significant difference in any outcome measure between PMMA injected into the CAJ and bilaterally abducted specimens (Controls 1 and 2) for either condition. The rate of increase in strain with increasing translaryngeal pressure was significantly less on the suture with PMMA placed in the CAJ (P = 0.03). CONCLUSIONS: These data provide strong evidence that injecting PMMA into the CAJ resists the collapsing effect of physiological airflows and pressures in vitro and reduces the force experienced by the laryngoplasty suture during maximal abduction. POTENTIAL RELEVANCE: Augmentation of prosthetic laryngoplasty with this technique may reduce arytenoid abduction loss in the early post operative period.

Racing performance following the laryngeal tie-forward procedure: a case-controlled study.

REASONS FOR PERFORMING STUDY: The laryngeal tie-forward procedure (LTFP) is becoming widely used for correction of dorsal displacement of the soft palate (DDSP) despite the absence of an evidence-based assessment of its efficacy. HYPOTHESES: The LTFP returns racing performance to preoperative baseline levels and to that of matched controls; and post operative laryngohyoid position is associated with post operative performance. DESIGN AND POPULATION: Case-controlled study of racehorses undergoing a LTFP for dorsal displacement of the soft palate at Cornell University between October 2002 and June 2007. METHODS: The presence of at least one post operative start and race earnings ($) were used as outcome variables. Controls were matched by age, breed and sex from the third race prior to surgery. A novel radiographic reference system was used to determine laryngohyoid position pre- and post operatively. Data for definitively and presumptively diagnosed cases were analysed separately. RESULTS: During the study interval, 263 racehorses presented, of which 106 were included in the study; 36 had a definitive diagnosis of DDSP and 70 a presumptive diagnosis. Treated horses were equally likely to race post operatively as controls in the equivalent race. Treated horses had significantly lower earnings in the race before surgery than matched controls. The procedure moved the basihyoid bone dorsally and caudally and the larynx dorsally and rostrally. A more dorsal post operative basihyoid position and more dorsal and less rostral laryngeal position were associated with an increased probability of racing post operatively. CONCLUSIONS: Horses undergoing a LTFP are as likely to race post operatively as matched controls. The procedure restores race earnings to preoperative baseline levels and to those of matched controls. POTENTIAL RELEVANCE: This study provides strong evidence supporting the use of the LTFP in racehorses. Further work is needed to determine the relationship between laryngohyoid conformation and nasopharyngeal stability in horses.

Racing performance after arthroscopic removal of apical sesamoid fracture fragments in Thoroughbred horses age < 2 years: 151 cases (1989--2002).

REASONS FOR PERFORMING STUDY: Studies on arthroscopic removal of apical proximal sesamoid fracture fragments in Thoroughbred (TB) horses age > or = 2 years have reported a high success rate. However, there are no reports documenting the racing prognosis of TB horses that undergo such surgery as weanlings or yearlings. OBJECTIVES: To describe the incidence of apical proximal sesamoid fractures in immature TB horses, age < 2 years and determine probability and quality of racing performance after arthroscopic removal of such fractures in 151 TB weanlings and yearlings. METHODS: The medical records of TB horses age < 2 years that underwent arthroscopic surgery for removal of apical proximal sesamoid fracture fragments were reviewed. Follow-up information was obtained from race records. Student's t tests were used to compare performance variables of operated racehorses to that of their maternal siblings. RESULTS: Ninety-two percent (139/151) of fractures occurred in the hindlimbs and 8% (11/151) in the forelimbs (fracture of both fore- and hindlimb, n = 1). Horses with forelimb fractures had a greatly reduced probability of racing (55%) compared to those with hindlimb fractures (86%). Overall, 84% of the horses raced post operatively and had performance records similar to that of their maternal siblings, 78% (787/1006) of which raced. CONCLUSIONS: Arthroscopic removal of apical proximal sesamoid fracture fragments in TB weanlings and yearlings carries an excellent prognosis for racing in horses with hindlimb fractures and a reduced prognosis in those with forelimb fractures. Medial fractures of the forelimb carry the worst prognosis. POTENTIAL RELEVANCE: The determination of prognosis increases knowledge on apical sesamoid bone fractures and potential for arthroscopic restoration of the ability to race; and enables the value of yearlings for subsequent sale to be established.

Plasma adrenocorticotropin, cortisol, and adrenocorticotropin/cortisol ratios in septic and normal-term foals.

BACKGROUND: Little information exists on the hypothalamic-pituitary-adrenal axis in septic foals. HYPOTHESIS: The plasma concentrations of adrenocorticotropin (ACTH) and cortisol are expected to be higher in septic foals as compared to normal foals. The concentrations of hormones in septic foals also are expected to differ further depending upon survival. ANIMALS: Twenty-eight control foals and 46 septic foals <14 days of age were included in this study. METHODS: Blood was collected in EDTA once from 28 normal foals born in the University of Georgia or Cornell University equine research herds and from 46 septic foals within 12 hours after admission to 1 of the 3 tertiary care referral centers involved in the study. Septic foal selection was based on a sepsis score of >11 or a positive blood culture. The control foals were age matched to the septic foals in the study. ACTH and cortisol concentrations were measured by a chemiluminescent immunoassay system. RESULTS: Cortisol concentrations in control foals did not vary with age. Septic foals had significantly higher mean ACTH, cortisol, and ACTH/cortisol ratios than did normal foals. Within the septic foal group, 28 foals survived to discharge, and 18 were euthanized or died. The mean age was not significantly different between the septic surviving and nonsurviving foals. The mean ACTH/cortisol ratio was significantly higher in the septic nonsurviving foals as compared to the septic surviving foals. CONCLUSIONS AND CLINICAL IMPORTANCE: Septic foals had higher hormone concentrations as compared to normal foals, which is an expected endocrine response to critical illness. The increased ACTH/cortisol ratio in nonsurviving septic foals in comparison to surviving septic foals could indicate hypothalamic-pituitary-adrenal axis dysfunction at the level of the adrenal gland in critically ill septic foals.

Risk factors associated with Cryptosporidium infection on dairy farms in a New York State watershed.

A cross-sectional study was carried out to determine the prevalence of Cryptosporidium parvum-like oocyst shedding on dairy farms in a watershed in New York State and to identify the factors that put animals at risk. A proportional sample of dairy herds in the targeted area was obtained, and animals were selected using a stratified sampling design to ensure representation of the population at risk. Fecal samples were collected per rectum and analyzed for the presence of C. parvum-like oocysts using the quantitative centrifugation concentration flotation technique and a proprietary enzyme-linked immunoassay. Additionally, isolates of Cryptosporidium were examined via bidirectional DNA sequencing. Data on putative risk factors were collected at the time of sampling and analyzed for association using logistic regression. The herd prevalence was 42% and the overall animal prevalence was 3.2%. The prevalence among animals less than 60 d of age was 20%. The likelihood of shedding Cryptosporidium decreased with the age of the animal and varied with the type of barn water source. Both the number of unweaned calves present at the time of the study, and whether the calves were tied vs. not tied increased the risk of infection. There was significant agreement between the flotation and PCR techniques. Sequencing revealed that 50% of the isolates were Cryptosporidium bovis, an isolate thought to be nonzoonotic.

Racing performance after arthroscopic removal of apical sesamoid fracture fragments in Thoroughbred horses age > or = 2 years: 84 cases (1989-2002).

REASONS FOR PERFORMING STUDY: Studies have shown that surgical removal of apical fracture fragments in Standardbred racehorses carries the best prognosis for return to racing performance, but there are no reports involving mature Thoroughbred (TB) racehorses. OBJECTIVES: To describe the incidence of apical proximal sesamoid fractures in TB racehorses and determine probability and quality of racing performance after arthroscopic removal of such fractures in TB racehorses age > or = 2 years. METHODS: Medical records and pre- and post operative race records of TB racehorses age > or = 2 years that underwent arthroscopic surgery for removal of apical proximal sesamoid fracture fragments were reviewed. RESULTS: Sixty-four percent of fractures occurred in the hindlimbs and 36% in the forelimbs. Horses with forelimb fractures had a reduced probability of return to racing (67%) compared to those with hindlimb fractures (83%), but the majority (77%) of treated horses recovered to return to race post operatively. Horses with medial forelimb fractures raced at only a 47% rate; those with suspensory desmitis at 63%. Unlike Standardbreds, there was no difference in probability of racing post operatively between horses that had, and had not, raced preoperatively. CONCLUSIONS: Data show that arthroscopic removal of apical proximal sesamoid fracture fragments is successful at restoring ability to race in skeletally mature TB horses without evidence of severe suspensory ligament damage. Prognosis for return to racing is excellent (83%) in horses with hindlimb fractures and good (67%) in those with forelimb fractures. Medial fractures of the forelimb have the worst prognosis. POTENTIAL RELEVANCE: The determination of prognosis for differing sites in TB racehorses should increase knowledge of apical proximal sesamoid bone fractures and improve communication from veterinarian to owner, and trainer, on the potential for arthroscopic restoration of the ability to race.

Quantitative risk assessment of Cryptosporidium species infection in dairy calves.

Cryptosporidium parvum is a zoonotic protozoan that infects many different mammals including cattle and humans. Cryptosporidiosis has become a concern for dairy producers because of the direct losses due to calves not performing well and the potential for environmental contamination with C. parvum. Identifying modifiable control points in the dynamics of infection in dairy herds will help identify management strategies that mitigate its risk. The quantitative risk assessment approach provides estimates of the risk associated with these factors so that cost-effective strategies can be implemented. Using published data from epidemiologic studies and a stochastic approach, we modeled the risk that C. parvum presents to dairy calves in 2 geographic areas: 1) the New York City Watershed (NYCW) in southeastern New York, and 2) the entire United States. The approach focused on 2 possible areas of exposure--the rearing environment and the maternity environment. In addition, we evaluated the contribution of many risk factors (e.g., age, housing, flies) to the end-state (i.e., total) risk to identify areas of intervention to decrease the risk to dairy calves. Expected risks from C. parvum in US dairy herds in rearing and maternity environments were 41.7 and 33.9%, respectively. In the NYCW, the expected risks from C. parvum in the rearing and maternity environments were 0.36 and 0.33%, respectively. In the US scenarios, the immediate environment contributed most of the risk to calves, whereas in the NYCW scenario, it was new calf infection. Therefore, within the NYCW, risk management activities may be focused on preventing new calf infections, whereas in the general US population, cleaning of calf housing would be a good choice for resource allocation. Despite the many assumptions inherent with modeling techniques, its usefulness to quantify the likelihood of risk and identify risk management areas is illustrated.

Age-related changes in serum insulin-like growth factor-I, insulin-like growth factor-I binding protein-3 and articular cartilage structure in Thoroughbred horses.

REASONS FOR PERFORMING STUDY: Structural changes in articular cartilage associated with the ageing process require definition for investigators performing developmental and age-related studies, for which information is lacking. OBJECTIVES: To 1) determine the onset and end of puberty as defined by serum insulin like growth factor (IGF-I) and IGF-binding protein-3 (IGFBP-3) concentrations and 2) correlate articular-epiphyseal cartilage complex structural changes with the onset and end of puberty. METHODS: IGF-I and IGFBP-3 were measured in serum samples from normal female and male horses age 9-715 days to determine peak and steady-state values for horses transitioning through puberty. Osteochondral tissue sections were obtained from horses age 120-840 days (4-28 months) and examined histologically for cartilage canals and tidemark formation. RESULTS: In male and female horses, serum IGF-I/IGFBP-3 concentrations peaked at approximately 225 days, defining the onset of puberty. Cartilage canals were absent from articular cartilage just prior to this time point. IGF-I/IGFBP-3 concentrations declined to steady-state levels at approximately age 450 days, signalling exit from puberty and therefore the beginning of ageing. This time point correlated to initial formation of a tidemark in the osteochondral tissue sections. CONCLUSIONS: Horses may be considered pubescent at age 225-450 days, and post pubescent and ageing after age 450 days. POTENTIAL RELEVANCE: Defining the normal post natal to post pubescent concentrations for serum IGF-I and serum IGFBP-3 establishes subsets of animals for age-related studies and may be used to monitor horses for abnormally high IGF-I concentrations due to natural disease or subsequent to systemic growth hormone administration.

Equine motor neuron disease is not linked to Cu/Zn superoxide dismutase mutations: sequence analysis of the equine Cu/Zn superoxide dismutase cDNA.

The cDNA encoding the equine copper/zinc superoxide dismutase (SOD1) was cloned from leukocyte total RNA from healthy horses and its nucleotide (nt) sequence was determined. We further sequenced the SOD1 gene from 16 horses diagnosed with equine motor neuron disease (EMND) and eight unrelated, clinically normal horses to determine if this disease, similar to amyotrophic lateral sclerosis (ALS) in humans, is linked to SOD1 mutations. The 465-bp SOD1 coding region in the horse encodes 153 amino acid (aa) residues. Equine SOD1 exhibited 81.8 and 79.9% sequence identity to the human homolog at the nt and aa levels, respectively, with only five distinct aa in the two loops that constitute the active site of the enzyme. None of the human SOD1 mutations found in the familial form of ALS were detected in SOD1 of the 16 affected horses. Although DNA sequence analysis identified three potential polymorphisms in equine SOD1, these were silent and were found in both normal and EMND-afflicted horses. At this time, there is no conclusive evidence for EMND linkage to SOD1 mutations.

Risk factors associated with equine motor neuron disease: a possible model for human MND.

Equine motor neuron disease (EMND), a newly described neurodegenerative disease, bears a striking resemblance to progressive muscular atrophy (PMA) in humans. We present a comparison of the equine and human diseases and the results of a case-control study conducted to identify intrinsic factors associated with EMND. Cases included all horses with a confirmed diagnosis of EMND diagnosed in the United States since 1985 (32 cases). Controls included horses diagnosed with either cervical stenotic myelopathy, equine degenerative myeloencephalopathy, or protozoan myelitis at the Veterinary Teaching Hospital at the College of Veterinary Medicine, Cornell University (153 controls). Logistic regression analysis identified factors associated with the risk of EMND. Risk factors considered were age, sex, and breed of the horse. Most cases of EMND (30 of 32) have been sporadic. There was a breed association with the risk of EMND. Quarter horses were at a high risk for developing EMND (odds ratio [OR] = 12.7; 95% confidence interval, 3.3 to 49.6); thoroughbred horses were at increased risk (OR = 2.9, 0.8 to 10.4). There was also an age association with the risk of EMND. The risk increased with age, peaked at 16 years, and then declined, a pattern similar to that for amyotrophic lateral sclerosis in humans. There was no sex association with the disease. Despite the breed association, equine lymphocyte antigen studies have not revealed a systematic pattern, suggesting that genetic factors influencing susceptibility to EMND may be outside the major histocompatibility complex.

Coordinate upregulation of cartilage matrix synthesis in fibrin cultures supplemented with exogenous insulin-like growth factor-I.

The addition of insulin-like growth factor-I to cartilage cultures is known to stimulate the synthesis of proteoglycan and type-II collagen in explant and monolayer studies. The purpose of this study was to determine the effects of long-term supplementation with insulin-like growth factor-I in chondrocytes cultured in fibrin discs as a preliminary investigation to in vivo application of chondrocyte/insulin-like growth factor-I/fibrin grafts to articular-cartilage repair procedures. Chondrocyte-fibrin cultures were maintained for 14 days, with insulin-like growth factor-I added at varying concentrations of 0, 10, 50, or 100 ng/ml medium. Cultures supplemented with 50 or 100 ng of growth factor/ml had increased levels of aggrecan and type-IIB procollagen mRNA, and translation to aggrecan and type-IIB collagen was confirmed by dye-binding assay of total proteoglycan, type-II collagen immunohistochemistry, and determination of collagen content by high-performance liquid chromatography. Maintenance of the chondrocyte phenotype during the 14 days of culture was confirmed by round cell morphology on routine staining, expression of type-II procollagen mRNA on in situ hybridization, evidence of production of pericellular type-II collagen on immunocytochemistry, synthesis of large-molecular-size aggrecan monomer on CL-2B column chromatography, and lack of appreciable message expression for type I or IIA collagen on Northern blot hybridization. Dose-response effects of insulin-like growth factor-I on the expression of chondrocyte matrix constituents were most pronounced at 50 and 100 ng of growth factor per milliliter of medium. These data confirm that (a) culture of chondrocytes for extended periods in three-dimensional cultures of fibrin maintains the chondrocyte phenotype and (b) supplementation with increasing concentrations of insulin-like growth factor-I enhances chondrocyte matrix synthesis and may provide a means to enhance chondrocyte phenotypic stability and function during transplantation grafting procedures.