Cytokine gene signatures in neural tissue of horses with equine protozoal myeloencephalitis or equine herpes type 1 myeloencephalopathy.

This study was designed to determine the relative levels of gene transcription of selected pathogens and cytokines in the brain and spinal cord of 12 horses with equine protozoal myeloencephalitis (EPM), 11 with equine herpesvirus type 1 (EHV-1) myeloencephalopathy, and 12 healthy control horses by applying a real time pcr to the formalin-fixed and paraffin-embedded tissues. Total rna was extracted from each tissue, transcribed to complementary dna (cDNA) and assayed for Sarcocystis neurona, Neospora hughesi, EHV-1, equine GAPDH (housekeeping gene), tumour necrosis factor (TNF)-alpha, interferon (IFN)-gamma, interleukin (IL)-1beta, IL-2, IL-4, IL-6, IL-8, IL-10 AND IL-12 p40. S neurona cdna was detected in the neural tissue from all 12 horses with EPM, and two of them also had amplifiable cDNA of N hughesi. The relative levels of transcription of protozoal cdna ranged from 1 to 461 times baseline (mean 123). All the horses with ehv-1 myeloencephalopathy had positive viral signals by PCR with relative levels of transcription ranging from 1 to 1618 times baseline (mean 275). All the control horses tested negative for S neurona, N hughesi and EHV-1 cdna. The cytokine profiles of each disease indicated a balance between pro- and anti-inflammatory markers. In the horses with epm the pro-inflammatory Th1 cytokines (IL-8, TNF-alpha and IFN-gamma) were commonly expressed but the anti-inflammatory Th2 cytokines (IL-4, IL-6 AND IL-10) were absent or rare. In the horses with ehv-1 the proinflammatory cytokine IL-8 was commonly expressed, but IL-10 and IFN-gamma were not, and TNF-alpha was rare. Tissue from the control horses expressed only the gene GAPDH.

Pathologic and bacteriologic findings in 27-week-old commercial laying hens experimentally infected with Salmonella enteritidis, phage type 4.

Two strains of 27-wk-old commercial laying chickens (strain A, brown-egg-laying type and strain B, white-egg-laying type) were inoculated either orally (PO) or intravenously (IV) with a field isolate of Salmonella enteritidis phage type 4. Chickens were sequentially necropsied at regular intervals throughout the 17-wk observation period. Gross and microscopic lesions were most evident between 1 and 14 days postinoculation (DPI). Gross lesions consisted of enlarged livers with white foci, enlarged and mottled white spleens, fibrinous exudate in the peritoneum, and atretic, misshapen ovarian follicles. Microscopic lesions included multifocal coagulative necrosis of hepatocytes and inflammation, fibrinous exudation in vascular sinuses of the spleen, and fibrinosuppurative inflammation of the peritoneum and ovarian follicles. The proportion of reproductive organ infections (ovary and oviduct) in the IV group, 83% (20/24, P = 0.007; 50% and 33% for strains A and strain B birds, respectively), was higher than that of the PO group, 46% (11/24; 29% and 17% for strains A and B, respectively), for the first 16 days of observation postinoculation. The proportion of fecal shedding for the IV group of birds was significantly (P = 0.009) lower, 29% (7/24; 33% and 25% respectively for strain A and strain B birds, respectively), than the PO group, 67% (16/24; 75% and 58% for strain A and strain B birds, respectively). Three (2.6%) of 234 egg pools were culture-positive for group D Salmonella from strain A chickens (1 of 119 pools from the IV group and 2 of 115 pools from the PO group of birds). Chickens infected with the field strain of S. enteritidis phage type 4 harbored the organism in tissues only for a brief time, most clearing within 8 DPI and nearly all within 16 DPI. Overall the percentage of culture-positive birds did not differ significantly (P > 0.05) between birds with and without lesions, but isolation of S. enteritidis tended to be more frequent when lesions were evident. This experiment also demonstrated that brown-egg-laying-type chickens were more susceptible than white-egg-laying-type chickens to S. enteritidis phage type 4 isolated from California based on gross and microscopic lesions and bacteriologic findings.

Hoof size, shape, and balance as possible risk factors for catastrophic musculoskeletal injury of Thoroughbred racehorses.

OBJECTIVE: To evaluate hoof size, shape, and balance as risk factors for catastrophic musculoskeletal injuries (CMI), including suspensory apparatus failure (SAF) and cannon bone condylar fracture (CDY) in Thoroughbred racehorses. ANIMALS: 95 Thoroughbred racehorses that died between 1994 and 1996. PROCEDURE: 38 quantitative measures of hoof size, shape, and balance were obtained from orthogonal digital images of the hoof and were compared between case horses with forelimb CMI (70), SAF (43), and CDY (10) injuries and control horses whose death was unrelated to the musculoskeletal system (non-CMI, 25). Comparison of group means between cases and controls was done using ANOVA, and multivariable logistic regression was used to estimate odds ratios. RESULTS: Odds of CMI were 0.62 times lower for a 5mm increase in ground surface width difference and 0.49 times lower for a 100-mm2 increase in sole area difference. Odds of SAF were 6.75 times greater with a 10 degrees increase in toe-heel angle difference and 0.58 times lower with a 100-mm2 increase in sole area difference. Odds of CDY were 0.26 times lower with a 3 degrees increase in toe angle, 0.15 times lower with a 5-mm increase in lateral ground surface width, and 0.35 times lower with a 100-mm2 increase in sole area difference. CLINICAL RELEVANCE: Decreasing the difference between toe and heel angles should decrease risk of SAF for Thoroughbred racehorses and should be considered in addition to increasing toe angle alone to help prevent catastrophic injury. Trimming the hoof to perfect mediolateral symmetry may not be a sound approach to avoiding injury.

Proliferative enterocolitis associated with dual infection with enteropathogenic Escherichia coli and Lawsonia intracellularis in rabbits.

Both enteropathogenic Escherichia coli (EPEC) and an obligate intracellular bacterium, previously referred to as an intracellular Campylobacter-like organism and now designated Lawsonia intracellularis, have been reported as causes of enterocolitis in rabbits. An outbreak of enterocolitis in a group of rabbits, characterized by an unusually high rate of mortality, was found to be associated with dual infection with EPEC and L. intracellularis. The EPEC strain was found to have eaeA gene homology but was negative for afrA homology. The absence of the afrA gene, which encodes the structural subunit for the AF/R1 pilus, indicates that this rabbit EPEC strain is distinct from the prototypic RDEC-1 strain. This finding suggests that rabbit EPEC strains widely reported in Western Europe, which lack AF/R1 pili, are also present in rabbits in the United States. Dual infection with these two pathogens in rabbits has not been previously reported and may have contributed to the unusually high mortality observed in this outbreak.

Relationship between race start characteristics and risk of catastrophic injury in thoroughbreds: 78 cases (1992).

OBJECTIVE: To investigate relationships of several racehorse characteristics and race conditions with risk of a catastrophic musculoskeletal injury (CMI) resulting in euthanasia in Thoroughbreds during racing in California in 1992. DESIGN: Retrospective longitudinal study. ANIMALS: Thoroughbreds that incurred CMI during racing and all California race entrants in 1992. PROCEDURE: Necropsy records were reviewed, and race start information was obtained. Incidence risk of CMI/1,000 race entrants was estimated. Relationships between CMI during racing and race-meet, entrant age and sex, race type and length, and racing surface type and condition were evaluated by use of logistic regression. RESULTS: Incidence risk of CMI was 1.7/1,000 entrants. A higher risk of CMI was found at 2 fair race-meets, with incidence risks of 4.9 and 5.5/1,000 entrants. Risk of injury in male horses was 1.7 times greater than that in female horses, and influence of age on risk depended on race type. Risk of injury for horses 2 to 5 years old was two times greater for claiming horses than for maiden horses. Race length or racing surface type (dirt vs turf) or condition (fast, muddy, yielding) were not significantly associated with risk of CMI. CLINICAL IMPLICATIONS: Incidence of CMI was similar among 12 of 14 major and fair race-meets and among various race lengths and racing surface types and conditions, whereas incidence of CMI was influenced by entrant age and sex as well as race type. Investigators should consider controlling for age and sex, race-meet, and race type whenever possible in studies of risk of CMI.

Fatal musculoskeletal injuries incurred during racing and training in thoroughbreds.

OBJECTIVE: To characterize and contrast data from Thoroughbreds that incurred a fatal musculoskeletal injury (FMI; injury resulting in death or euthanasia) during racing or training and data from all California race entrants during a 9-month period in 1991. DESIGN: Case-control study. ANIMALS: Thoroughbreds that incurred a FMI during racing or training at a California race-meet and all California race entrants from January through June and October through December 1991. PROCEDURE: Age and sex were compared with chi 2 and Fisher's exact tests among horses fatally injured while racing and training. A log-linear model was fit to assess the relationship between race-meet and age and sex of California race entrants. Incidence risk of racing FMI was estimated per 1,000 race entrants, and the relationship between the occurrence of FMI during racing with race-meet, age, and sex was evaluated by logistic regression. RESULTS: Injury type and sex-specific age distributions differed among the horses fatally injured during racing and training. Age and sex distributions of the race entrants were not independent and varied among race-meets. Overall incidence risk of racing FMI was estimated at 1.7/1,000 race entrants. Risk of racing FMI in male horses was about twofold that in female horses, and in 4-year-olds was twofold that in 3-year-olds. CLINICAL IMPLICATIONS: Age and sex-related differences in risk of incurring a FMI during racing should be considered when comparing fatal injury rates among race-meets.

Causes of death in racehorses over a 2 year period.

Necropsies were performed on 496 horses that had a fatal injury or illness at a California racetrack during the period February 20th 1990 to March 1st 1992. The primary cause of death was categorised by breed, activity at time of injury or illness and organ system affected. Most of the submissions were Thoroughbred horses (432) and Quarter Horses (46). Most of the injuries occurred while racing (42%) and in training sessions (39%); with fewer non-exercise (12%) and accident (7%) related injuries or illnesses. Musculoskeletal injuries accounted for 83% of the Thoroughbred and 80% of the Quarter Horse submissions. The Thoroughbred horses incurred 306 fractures with 263 in the limbs and 90% of those in the forelimbs. The proximal sesamoid bone(s), third metacarpal bone and humerus were the most common bones fractured in Thoroughbred horses and Quarter Horses. Other major causes of death included respiratory, digestive and multi-organ system disorders.

A retrospective study of circovirus infection in pigeons: nine cases (1986-1993).

Circovirus infections were diagnosed in 12 pigeons from the United States 4 pigeons from Australia, and 1 pigeon from Canada (1986-1993). Circovirus was identified by electron microscopic examination of basophilic botryoid cytoplasmic inclusions that had a histologic appearance similar to that of psittacine beak and feather disease virus inclusions. Inclusions were seen in splenic, bursal, gut-associated, and bronchus-associated lymphoid tissue macrophages and in bursal epithelial cells. Inclusions were composed of paracrystalline arrays of tightly packed, nonenveloped icosahedral virions 14-17 nm in diameter. Histologic changes in the spleens ranged from lymphofollicular hyperplasia with mild discrete lymphocellular necrosis to lymphoid depletion and diffuse histiocytosis. Lesions in the bursa of Fabricius ranged from mild lymphocellular necrosis to severe cystic bursal atrophy. Remaining histologic findings coincided with concurrent bacterial, viral, fungal, and parasitic infections. Immunoperoxidase staining and DNA in situ hybridization demonstrated that pigeon circovirus is distinct from psittacine beak and feather disease virus; however both viruses apparently share some homologous DNA sequences. Clinical and diagnostic findings indicate that pigeon circovirus may be similar to psittacine beak and feather disease virus with respect to acquired immunodeficiency and subsequent multiple secondary infections.

Mycoplasma bovis associated with decubital abscesses in Holstein calves.

Between April of 1990 and March of 1992, calves on a Holstein calf ranch experienced subcutaneous decubital abscesses involving the brisket region, dorsal aspect of the carpus, and lateral aspect of the stifle joints. Fifty out of 2,500 (2%) Holstein calves between the ages of 3 and 12 weeks were affected. Needle aspirates of brisket abscesses from 8 calves and 6 live or dead calves with 1 or more decubital abscesses were submitted for examination. Two of the 6 calves in addition had bronchopneumonia. Mycoplasma bovis was isolated from all abscesses and 1 lung. Formalin fixed tissues taken from the affected areas also revealed M. bovis by immunoperoxidase staining. No evidence of joint involvement was apparent, and no mycoplasma was isolated from the joints adjacent to affected areas. Attempts to isolate mycoplasma from milk and environmental samples were unsuccessful.

Intranuclear inclusions of unknown pathogenic significance from diagnostic cases in three avian species.

Intranuclear inclusions were observed with light microscopy in tissues from necropsy cases from three different species of birds. Because of the nature of these inclusions, the species affected, or their distribution, the inclusions were considered of unknown pathogenic significance. The inclusions were examined ultrastructurally. Parvovirus-like particles were observed in the intranuclear inclusions of pigeon hepatocytes and duodenal stromal cells from a quail. Finely granular eosinophilic intranuclear inclusion bodies from two unrelated pigeon cases were found to be composed of loosely organized filaments. One liver sample from a 16-week-old laying chicken had prominent eosinophilic hepatocellular intranuclear inclusions composed of electron-dense coarsely granular amorphous material.

Tracheal aspergillosis in 6 1/2-week-old chickens caused by Aspergillus flavus.

A case of localized tracheal aspergillosis in 6 1/2-week-old single-comb white leghorn pullets caused by Aspergillus flavus is documented. Yellow caseous plaques adherent to the mucosal surface of the tracheas were observed grossly. In several tracheas, the plaques occluded the lumina, and the surrounding tracheal walls were reddened. Histologically, the mucosa was necrotic and infiltrated with macrophages, and fibroplasia was evident in the subadjacent tracheal wall. The lumen of the trachea was almost completely occluded by a combination of fungal mycelia and pyogranulomatous exudate. Portions of tracheal cartilage were elevated into the lumen of the trachea. Other than a sudden increase in mortality to 0.5% per day, there was no evidence of disease in the flock. Depletion of bursal lymphocytes, with concomitant cryptosporidiosis, was evident on histological examination. Acute infectious bursal disease was diagnosed in the succeeding flock at this ranch based upon serology and typical histology.

An association between complete and incomplete stress fractures of the humerus in racehorses.

Twenty-one horses had a complete unilateral humeral fracture during race training or racing at a California racetrack during the period 24 February 1990 to 10 July 1991. Fractures occurred approximately equally in left and right limbs, and in males and females. Most fractures occurred during training, and in 2- and 3-year-old horses. Only 5 of 16 Thoroughbred horses with known racing records had previously raced more than once, and their mean time between races was less than the time between their last race and fracture (P = 0.07). Ten of 13 humeri studied further had gross evidence of periosteal callus bridging one portion of the fracture line, indicative of a pre-existing stress fracture.

Actinomycetales infections in slender-horned gazelles: six cases (1987-1989).

Disseminated Actinomycetales infections were diagnosed in 6 slender-horned gazelles at a zoologic park over a 17-month period. Nocardia and Mycobacterium spp were isolated. Possible predisposing causes leading to infection were investigated. Environmental contamination where the gazelles were housed was not high, and other breeds of gazelles at the park did not become infected, indicating that environmental exposure was not the sole factor involved. Information gathered from questionnaires sent to other zoologic parks and personnel communications indicated that this was not an isolated incident. Investigation into the genetic lineage of the gazelles revealed substantial inbreeding in prior generations. These findings suggested inbreeding may have been an important predisposing factor leading to infection. Careful scrutiny to ensure maximal heterozygosity of future breedings is warranted.

Meningoencephalitis in commercial meat turkeys associated with Mycoplasma gallisepticum.

Mycoplasma gallisepticum (MG) infection was diagnosed in three different flocks of 12-to-16-week-old commercial meat turkeys displaying torticollis and/or opisthotonos. MG was isolated from the brain, air sacs, trachea, and sinus of one bird with neurological signs. Histological examination of brains in all three cases revealed moderate-to-severe encephalitis with lymphoplasmacytic cuffing of vessels, fibrinoid vasculitis, focal parenchymal necrosis, and meningitis. Birds with neurological signs were seropositive for MG by the serum-plate agglutination and hemagglutination-inhibition tests. The encephalitic form of MG has been described previously but is rarely mentioned in the current literature.

Clostridium botulinum type-C intoxication associated with consumption of processed alfalfa hay cubes in horses.

An episode of nervous system dysfunction was observed in horses on 17 premises in 4 counties of southern California. Thirty-eight horses were affected, and 31 of those died. The common clinical signs of disease in the affected horses were: increased appetite; anxious attitude; rythmic, intermittent muscle tremors in the area of the tricep muscles; decreased palpebral tone; mydriasis; small hard fecal balls; and tendency to become sternally recumbent with the neck extended. The temporal distribution of cases on all 17 premises suggested a relationship between exposure to a common batch of alfalfa hay cubes and manifestations of similar clinical signs of disease in affected horses. Fifteen horses were submitted for necropsy. Diagnosis of botulism was established on the basis of detection of type-C1 toxin in the feed, in intestinal contents of 1 horse, and in the liver of the aforementioned horse and another horse. Toxigenic strains of Clostridium botulinum type-C were isolated from intestinal contents of 5 affected horses, one of which also contained type-C1 and type-C2 toxins. Seven of 10 horses treated with type-C antitoxin and plasma obtained from horses hyperimmunized with C botulinum type-C toxoids survived.

Viral pathogenesis of a nephrotropic infectious bronchitis virus isolated from commercial pullets.

Infectious bronchitis was diagnosed in 3-to-4-week-old pullets from an outbreak in a commercial flock in California. The disease was characterized by head swelling, watery discharge from the eyes and nostrils, and urates in kidneys. Mortality ranged from 1.8% to 12.5% per week. The isolation of a coronavirus from a suspension of pooled kidneys from clinically ill chickens at the fifth passage in 10-day-old chicken embryos, gross and histologic renal lesions, and seroconversion by enzyme-linked immunosorbent assay in inoculated birds suggested that the virus isolated was a nephrotrophic strain of infectious bronchitis virus (IBV). The virus isolate was found to be a previously unrecognized serotype, based on virus neutralization tests performed in embryonated chicken eggs. Nephropathogenicity of the IBV isolate was confirmed by inoculation of the viral isolate into specific-pathogen-free chicks and demonstration of renal lesions. The isolation of nephrotropic strains of IBV has not been reported previously from poultry in California.

Bovine fetal encephalitis and myocarditis associated with protozoal infections.

Bovine fetuses submitted to the California Veterinary Diagnostic Laboratory System were evaluated during a 2-year period (1987 to 1989) for the presence of multifocal necrotizing nonsuppurative encephalitis, nonsuppurative myocarditis, or tissue protozoa. Eighty-two of 445 (18%) fetuses submitted met these histologic criteria. Fetuses were from 54 dairy and two beef herds located throughout the state. In 17 fetuses (21%) protozoa were found in fetal tissues. Protozoa were found in brain parenchyma of ten fetuses (12%), in endothelial cells in four fetuses (5%), in cardiac myofibers in one fetus (1%), and were associated with endothelial cells in two fetuses (2%). In most fetuses there were no significant gross pathologic findings other than autolysis. While aborted fetuses were from 3 to 9 months gestation, the majority were between 5 and 7 months gestation. They were submitted year round, but more were seen in the fall and winter months. Additional salient histologic features included portal nonsuppurative hepatitis, focal hepatic inflammation and necrosis, and focal nonsuppurative myositis. Nonsuppurative inflammation was also found in decreasing frequency, in the adrenal medulla, kidney, mesentery or abdominal fat, placenta, and lung. In two fetuses (Nos. 1 and 2), the location and morphology of the protozoa were compatible with Sarcocystis spp. The identity of protozoa in the remaining 15 fetuses is unknown. The histopathologic changes in these 82 fetuses and the presence of protozoa in 21% of the fetuses suggest these abortions are due to fetal protozoal infections.