Veterinary Forensic Pathology: The Search for Truth.

Veterinary forensic pathology is emerging as a distinct discipline, and this special issue is a major step forward in establishing the scientific basis of the discipline. A forensic necropsy uses the same skill set needed for investigations of natural disease, but the analytical framework and purpose of forensic pathology differ significantly. The requirement of legal credibility and all that it entails distinguishes the forensic from routine diagnostic cases. Despite the extraordinary depth and breadth of knowledge afforded by their training, almost 75% of veterinary pathologists report that their training has not adequately prepared them to handle forensic cases. Many veterinary pathologists, however, are interested and willing to develop expertise in the discipline. Lessons learned from tragic examples of wrongful convictions in medical forensic pathology indicate that a solid foundation for the evolving discipline of veterinary forensic pathology requires a commitment to education, training, and certification. The overarching theme of this issue is that the forensic necropsy is just one aspect in the investigation of a case of suspected animal abuse or neglect. As veterinary pathologists, we must be aware of the roles filled by other veterinary forensic experts involved in these cases and how our findings are an integral part of an investigation. We hope that the outcome of this special issue of the journal is that veterinary pathologists begin to familiarize themselves with not only forensic pathology but also all aspects of veterinary forensic science.

Nondrowning Asphyxia in Veterinary Forensic Pathology: Suffocation, Strangulation, and Mechanical Asphyxia.

Asphyxia in a forensic context refers to death by rapid cerebral anoxia or hypoxia due to accidental or nonaccidental injury. Death due to nondrowning asphyxia can occur with strangulation, suffocation, and mechanical asphyxia, each of which is categorized based on the mechanism of injury. Individuals dying due to various types of asphyxia may or may not have lesions, and even those lesions that are present may be due to other causes. The interpretation or opinion that death was due to asphyxia requires definitive and compelling evidence from the postmortem examination, death scene, and/or history. Beyond the postmortem examination, pathologists may be faced with questions of forensic importance that revolve around the behavioral and physiological responses in animals subjected to strangulation, suffocation, or mechanical asphyxia to determine if the animal suffered. While there is no prescriptive answer to these questions, it is apparent that, because of physiological and anatomical differences between humans and animals, for some mechanisms of asphyxia, consciousness is maintained for longer periods and the onset of death is later in animals than that described for people. Veterinary pathologists must be cognizant that direct extrapolation from the medical forensic literature to animals may be incorrect. This article reviews the terminology, classification, mechanisms, and lesions associated with asphyxial deaths in companion animals and highlights significant comparative differences of the response to various types of asphyxia in animals and people.

Veterinary Forensic Pathology: Drowning and Bodies Recovered From Water.

Determining the cause of death in animals recovered from bodies of water, swimming pools, or other water-containing vessels is challenging. Animals recovered from water may or may not have drowned. The diagnosis of drowning is usually one of exclusion, requiring information from the crime scene, recovery scene, the medical history or reliable witness accounts. While there are characteristic macroscopic and microscopic lesions of drowning, none are specific and are dependent on the volume and tonicity of the drowning medium. Beyond interpreting the postmortem findings, the court may ask pathologists to comment on the behavioral and welfare implications of drowning. This requires an understanding of the drowning process, which is a complex series of sequential, concurrent, and overlapping cardiorespiratory reflexes, electrolyte and blood gas abnormalities, aspiration, physical exhaustion, and breathlessness eventually culminating in death. This review addresses the mechanisms, lesions, and diagnostic issues associated with drowning in nonaquatic companion animals.

A Survey of Attitudes of Board-Certified Veterinary Pathologists to Forensic Veterinary Pathology.

An electronic survey was conducted to determine the attitudes of veterinary pathologists toward forensic pathology and the adequacy of their training in the discipline. The survey was sent to 1933 diplomates of the American College of Veterinary Pathologists and 311 completed responses were analyzed. Of respondents, 80% report receiving at least 1 type of medicolegal case, with cases from law enforcement received most frequently. Most (74%) of the respondents indicated that their previous training did not prepare them adequately to handle forensic cases and almost half of the respondents (48%) indicated that they needed more training on serving as an expert witness. Relative risk ratios (RRR) and odds ratios (OR) were generated to determine the strength of a statistically significant association. Responses from a free-text entry question determining additional training needs could be grouped into 3 main categories: (1) veterinary forensic pathology science and procedures, (2) documentation, evidence collection and handling, and (3) knowledge of the medicolegal system. Last, a field for additional comments or suggestions regarding veterinary forensic pathology was completed by 107 respondents and many reinforced the need for training in the categories previously described. The survey highlights that a significant proportion of diplomates of the American College of Veterinary Pathologists are currently engaged in veterinary forensic pathology but feel their training has not adequately prepared them for these cases. Hopefully, the survey results will inform the college and residency training coordinators as they address the training requirements for an important emerging discipline.

Syndromic surveillance using laboratory test requests: a practical guide informed by experience with two systems.

Syndromic surveillance systems can enhance early disease warning, endemic disease monitoring, or help to accumulate proof of disease freedom. In order to provide immediate feedback to achieve these goals, the health data sources scanned should be acquired continuously, in an automated fashion, and should be stored electronically. Recognizing that data from diagnostic test requests often meet these requirements, two systems designed to automatically extract surveillance information from animal laboratory databases have been developed and are described in this paper. These systems are designed to contribute to early disease detection, as well as the timely management of epidemiological information, in a province of Canada and in Sweden, the areas served by the diagnostic laboratories concerned. Classifying in-coming requests into syndromes, the first step, was the most time-consuming and the least portable step between the two systems. The remaining steps were more easily adjusted from one system to implementation in the other. These steps included: retrospective evaluation of data to create baseline profiles following the removal of excessive noise and aberrations; the identification of temporal effects; prospective evaluation of detection algorithms; and finally real-time monitoring and implementation. Building upon the institutions' existing data management software, all steps to use those data for the purposes of syndromic surveillance were set up using open source software; as a result this approach could be readily adopted by other institutions. Relatively straight-forward development and maintenance is expected to lead to the incorporation of these systems into each institution's surveillance processes, becoming an indispensable tool for diagnosticians and epidemiologists, as well as stimulating further technical development of such systems.

An investigation of the etiology of a mild diarrhea observed in a group of grower/finisher pigs.

An investigation into a mild diarrhea in a group of grower/finisher pigs was carried out in order to determine the etiology. A tiamulin injection and a carbadox-medicated ration were given to pens of pigs in a 2 x 2 factorial experimental design. Pens of pigs were assessed a score, based on the consistency of the feces in the pen, each week. The clinical investigation looked for the intestinal pathogens Brachyspira pilosicoli, B. hyodysenteriae, Lawsonia intracellularis, Salmonella spp., Yersinia spp., transmissible gastroenteritis virus, and rotavirus. Despite a rigorous investigation, the diarrhea was not attributed to any pathogen. A mild colitis was noted among pigs necropsied while affected with diarrhea. Improved diagnostic tools may allow a more effective response to an outbreak of mild disease, while at the same time reducing the amount of antimicrobials used in swine production.

Coxiella burnetii infection is associated with placentitis in cases of bovine abortion.

A positive score on a modified acid-fast (MAF)-stained smear test of fresh placenta was used to identify a group of bovine abortion submissions believed to be infected with Coxiella burnetii. Immunohistochemical (IHC) testing for Coxiella and Chlamydia antigens was performed on 14 MAF smear-positive cases as well as 29 MAF smear-negative cases received during the study period. Nine MAF smear-positive cases as well as 1 MAF smear-negative case were Coxiella-positive via the IHC test. No placentas were positive for Chlamydia antigen. Various histopathologic features were categorized for all placentas and the presence or absence of selected risk categories was also graded for each case. The results between Coxiella IHC-positive cases and Coxiella IHC-negative/MAF-negative cases were compared using Fisher's exact test (P value at 95% confidence). Significant associations were found between Coxiella IHC-positive cases and the presence of placental inflammation (P = 0.0027), placental necrosis (P = 0.012), fetal pneumonia (P = 0.0152), and the visibility of Coxiella-like organisms within trophoblasts on hematoxylin and eosin-stained sections (P < 0.0001). Histopathologic features of Coxiella IHC-positive placentas included infiltration of the chorionic stroma by mononuclear cells, necrosis of chorionic trophoblasts, and focal exudation of fibrin and neutrophils. The results indicate that MAF smears are a good screening tool for the presence of Coxiella in placentas from bovine abortion cases and that the detection of this pathogen in aborted placentas via traditional staining or IHC methods is usually associated with placentitis.

A descriptive study of the frequency and characteristics of proliferative enteropathy in swine in Ontario by analyzing routine animal health surveillance data.

Routine surveillance data, collected on pathology submissions at the Animal Health Laboratory in Guelph between 1992 and 1997, were analyzed to determine demographic, clinical, and pathologic characteristics of cases of proliferative enteropathy and the frequency of this condition relative to other infectious enteric diseases in swine in Ontario. The most commonly reported disease was Escherichia coli enteritis (average cases/year = 70.0). Among infectious enteropathies that occur typically in neonatal pigs, coccidiosis (28.4 cases/year) and rotaviral enteritis (5.6 cases/year) were reported. Among infectious enteropathies generally associated with diarrhea in weaner and grower/finisher pigs, the most frequently reported was proliferative enteropathy (27.6 cases/year), followed by swine dysentery (23.3 cases/year), transmissible gastroenteritis (19.6 cases/year), and salmonellosis (8.4 cases/year). Diarrhea and bloody diarrhea were reported in 29% and 31%, respectively, of herds diagnosed with proliferative enteropathy. Important gross intestinal lesions included mucosal hypertrophy (62% of cases), hemorrhage (47%), and mucosal necrosis (34%). Histologic intestinal lesions included epithelial hyperplasia (90% of cases), mucosal necrosis (59%), and inflammation (49%). Our results suggest that proliferative enteropathy is a major infectious enteric disease in grower/finisher pigs in Ontario.

Eosinophils: a review.

The eosinophil was discovered by Jones in 1846 (Dessein and David, 1982) but its proclivity to stain with aniline dyes was first described by Paul Ehrlich in 1879 (Hirsch and Hirsch, 1980). Recognized and named for this quality, eosinophils possess an abundance of highly basic proteins within their granules which confer their affinity for acidic dyes (Gleich and Loegering, 1984). Eosinophils are traditionally viewed as killer-effector cells in parasitic infestations and as modulators of Type I hypersensitivity reactions (Butterworth and David, 1981; Kay, 1985). The eosinophils' reserve of cationic proteins and enzymes which imparts their profound parasiticidal effects (Butterworth and David, 1981) contrasts with this leukocyte's purported regulatory function in inflammation (Kay, 1985; Fechter et al., 1986). The opposing functions possessed by this leukocyte exemplify the enigma of the eosinophil. Recent evidence suggests that although the eosinophil does posses some regulatory capabilities, its presence is, in fact, a harbinger of tissue destruction (Gleich and Adolphoson, 1986, Wardlaw and Kay, 1987; Spry, 1988). Nor does the presence of the eosinophil automatically infer IgE mediated hypersensitivity, as evidenced by studies examining the interaction of the eosinophil with the cellular arm of the immune system (Basten and Beeson, 1970; Ruscetti et al., 1976; Beeson and Bass, 1977; Raghavachar et al., 1987; Ohnishi et al., 1988). The purpose of this review is to provide a brief overview of the structure and biology of the mammalian eosinophil and to emphasize the fact that eosinophils fulfil a paradoxical role as effectors of tissue damage and as benign modulators of inflammation.

The effect of leukotriene B4, leukotriene C4, zymosan activated serum, histamine, tabanid extract and N-formyl-methionyl-leucyl-phenylalanine on the in vitro migration of equine eosinophils.

The migration of equine eosinophils under agarose in response to inflammatory mediators, an arthropod extract and a synthetic peptide was examined. A chemotactic index (CI) was calculated by determining the ratio of the distance of eosinophil migration towards the chemoattractant to the distance migrated towards a buffer. Differences between the CI of those eosinophils exposed to chemoattractants and those exposed only to buffer were assessed by an analysis of variance. All agents except leukotriene C4 and the buffer induced statistically significant directional migration of eosinophils. Leukotriene B4 (LTB4) was the most effective chemotaxin for equine eosinophils. Migration of eosinophils stimulated by 10(-9) M LTB4 exceeded that induced by concentrations of histamine six orders of magnitude greater. The response of equine eosinophils to inflammatory mediators was similar to the reported behavior of human eosinophils. The ability of tabanid extract to attract equine eosinophils suggests that arthropod induced tissue eosinophilia many not depend entirely upon immunological mechanisms. The peptide, N-formyl-methionyl-leucyl-phenylalanine attracted equine eosinophils at 10(-4) M and 10(-3) M, concentrations that exceed those reported to be stimulatory for eosinophils of other species. The results of this study indicate that equine eosinophils are capable of migrating towards diverse stimuli, of which LTB4 was the most effective. It is plausible that LTB4 figures prominently in equine inflammation, particularly in lesions dominated by eosinophils.

Auricular chondritis in Wistar rats.

The macroscopic and microscopic features of auricular chondritis in Wistar rats are described. Histologic lesions are characterized by granulomatous inflammation, chondrolysis, and multinodular proliferative foci. The condition in Wistar rats is compared to auricular chondritis in other strains of rats.