Conceptual modeling of postmortem evaluation findings to describe dairy cow deaths.

Dairy cow mortality levels in the United States are excessive and increasing over time. To better define cause and effect and combat rising mortality, clearer definitions of the reasons that cows die need to be acquired through thorough necropsy-based postmortem evaluations. The current study focused on organizing information generated from postmortem evaluations into a monitoring system that is based on the fundamentals of conceptual modeling and that will potentially be translatable into on-farm relational databases. This observational study was conducted on 3 high-producing, commercial dairies in northern Colorado. Throughout the study period a thorough postmortem evaluation was performed by veterinarians on cows that died on each dairy. Postmortem data included necropsy findings, life-history features (e.g., birth date, lactation number, lactational and reproductive status), clinical history and treatments, and pertinent aspects of operational management that were subject to change and considered integral to the poor outcome. During this study, 174 postmortem evaluations were performed. Postmortem evaluation results were conceptually modeled to view each death within the context of the web of factors influencing the dairy and the cow. Categories were formulated describing mortality in terms of functional characteristics potentially amenable to easy performance evaluation, management oversight, and research. In total, 21 death categories with 7 category themes were created. Themes included specific disease processes with variable etiologies, failure of disease recognition or treatment, traumatic events, multifactorial failures linked to transition or negative energy balance issues, problems with feed management, miscellaneous events not amenable to prevention or treatment, and undetermined causes. Although postmortem evaluations provide the relevant information necessary for framing a cow's death, a restructuring of on-farm databases is needed to integrate this level of detail into useful monitoring systems. Individual operations can focus on combating mortality through the use of employee training related to postmortem evaluations, detailed forms for capturing necropsy particulars and other relevant information related to deaths, and standardized nomenclature and categorization schemes. As much as anything, the simple act of recognizing mortality as a problem might be the most fundamental step toward controlling its progression.

A necropsy-based descriptive study of dairy cow deaths on a Colorado dairy.

Increasing levels of dairy cow mortality pose a challenge to the US dairy industry. The industry's current understanding of dairy cow mortality is reliant upon descriptions largely based on producer or veterinary assumptions regarding cause of death without the benefit of detailed postmortem evaluations. A thorough necropsy is a superior tool for establishing a cause of death, except for cases involving euthanasia for traumatic accidents or severe locomotor disorders. Information provided from a necropsy examination would be most valuable if it were categorized and combined with cow health information in a complete postmortem evaluation designed to guide future management decisions. The objective of this study was to describe dairy cow deaths on a Colorado dairy over a 1-yr period and explore classification systems for necropsy findings that might inform management actions aimed at reducing dairy cow mortality. Throughout the study period a thorough necropsy examination was performed on every cow that died. Based upon this examination each death was characterized by a proximate cause (i.e., the most likely immediate cause of the death). Each proximate cause of death was then categorized using 3 alternate schemes founded on generalized etiologic principles and influenced by previous clinical history and treatments. These schemes included the broad categories commonly used for classifying findings within a review of literature related to dairy cow mortality, a diagnostic scheme used within the problem-oriented veterinary medical record, and an analysis focusing on the primary physiologic system derangement for each death. A total of 2,067 cows were enrolled during the study period of which 1,468 cows freshened, 507 cows were sold, and 94 cows died, resulting in a mortality risk of 6.4 deaths per 100 lactations at risk. The distribution of deaths by parity was significantly different from the herd distribution at the end of study with the largest percentage of death present in parity > or =4. Postmortem findings attributable to a specific cause of death were present for all but 4 of the 94 deaths. Assignment of the proximate causes of death to categories within the 3 alternate schemes provided a means for classifying necropsy findings and causes of death with different levels of detail. Creating categories with more selective groupings may provide a means for capturing specifics related to deaths that can be used to guide management decisions.

Association of severity of enteric granulomatous inflammation with disseminated Mycobacterium avium subspecies paratuberculosis infection and antemortem test results for paratuberculosis in dairy cows.

Disseminated infection (DI) of Mycobacterium avium subspecies paratuberculosis (MAP) in cattle may impair cow health, potentiate spread of disease, and is a potential food-safety risk. The objectives of this study were to determine the association between severity of histologic enteric lesions and the occurrence of DI, clinical signs, and positive fecal culture and serum ELISA results. Bacteriologic fecal culture and serum ELISA were performed on 40 dairy cows from MAP-infected herds. Cows were classified as having DI if MAP was isolated from any of 11 extra-intestinal tissues collected postmortem. A grade of 0-3, corresponding to the severity of histologically evident granulomatous inflammation was determined for sections of ileum, jejunum, mesenteric lymph node, and ileocolic lymph node. An overall intestinal inflammation (OII) grade of 0-3 was assigned to each cow. The proportion of cows with DI increased with tissue-specific lesion grade and OII grade. All cows with grade 3 inflammation in any single tissue had DI, however, some cows with DI had grade 1 inflammation or no lesions. In general, there was a positive association between OII grade and clinical signs, gross enteric lesions, and positive ELISA and fecal culture results. However, 12% of OII grade 0 cows had clinical signs (explained by other conditions recognized with necropsy), and the proportion of positive ELISA results was lower for OII grade 3 cows relative to grade 2 cows. Although MAP dissemination may occur early in the disease process, histopathology of intestinal tissues may be used to detect a substantial proportion of DI cows.

Globoid cell-like leukodystrophy in a domestic longhaired cat.

Globoid cell leukodystrophy (GLD; Krabbe disease), is a rare heritable metabolic disorder in humans, dogs, mutant twitcher mice, and rhesus monkeys that is caused by a deficiency in the lysosomal enzyme galactocerebrosidase (GALC). GALC deficiency results in the accumulation of psychosine, which is toxic to oligodendrocytes and Schwann cells of the central and peripheral nervous systems. Clinical signs include hypotonia, mental regression, and death by 2 years of age in most human patients. Here we describe a domestic longhaired kitten with rapidly progressive neurologic disease and brain and spinal cord lesions characteristic of GLD. Pathologic hallmarks of the disease reflect the loss of oligodendrocytes and include myelin loss, gliosis, and the perivascular accumulation of large mononuclear cells with fine cytoplasmic vacuoles (globoid cells) in the peripheral and central nervous systems. Globoid cells were CD68 and ferritin positive, confirming their monocytic origin, and cytoplasmic contents were nonmetachromatic and periodic acid-Schiff positive.

Involvement of microbial respiratory pathogens in acute interstitial pneumonia in feedlot cattle.

OBJECTIVE: To evaluate viral and bacterial respiratory pathogens and Mycoplasma spp isolated from lung tissues of cattle with acute interstitial pneumonia (AIP) and cattle that had died as a result of other causes. SAMPLE POPULATION: 186 samples of lung tissues collected from cattle housed in 14 feedlots in the western United States. PROCEDURE: Lung tissues were collected during routine postmortem examination and submitted for histologic, microbiologic, and toxicologic examinations. Histologic diagnoses were categorized for AIP, bronchopneumonia (BP), control samples (no evidence of disease), and other disorders. RESULTS: Cattle affected with AIP had been in feedlots for a mean of 1272 days before death, which was longer than cattle with BP and control cattle. Detection of a viral respiratory pathogen (eg, bovine respiratory syncytial virus [BRSV], bovine viral diarrhea virus, bovine herpesvirus 1, or parainfluenza virus 3) was not associated with histologic category of lung tissues. Bovine respiratory syncytial virus was detected in 8.3% of AIP samples and 24.0% of control samples. Histologic category was associated with isolation of an aerobic bacterial agent and Mycoplasma spp. Cattle with BP were at greatest risk for isolation of an aerobic bacterial agent and Mycoplasma spp. CONCLUSIONS AND CLINICAL RELEVANCE: Analysis of these results suggests that AIP in feedlot cattle is not a consequence of infection with BRSV. The increased, risk of isolation of an aerobic bacterial agent from cattle with AIP, compared with control cattle, may indicate a causal role or an opportunistic infection that follows development of AIP.

Association of 3-methyleneindolenine, a toxic metabolite of 3-methylindole, with acute interstitial pneumonia in feedlot cattle.

OBJECTIVE: To compare concentrations of 3-methyleneindolenine (3MEIN) in lung tissues obtained from feedlot cattle that died as a result of acute interstitial pneumonia (AIP) and cattle that died as a result of other causes and to compare blood concentrations of 3MEIN in healthy feedlot cattle and feedlot cattle with AIP. STUDY POPULATION: Blood samples and lung tissues collected from 186 cattle housed in 14 feedlots in the western United States. PROCEDURE: Samples of lung tissues were collected during routine postmortem examination and submitted for histologic, microbiologic, and toxicologic examination. Blood samples were collected from cattle with clinical manifestations of AIP and healthy penmates. Histologic diagnoses were categorized as AIP, bronchopneumonia (BP), control samples, and other disorders. Concentrations of 3MEIN were determined in lung tissues and blood samples, using an ELISA. RESULTS: Concentrations of 3MEIN in lung tissues were significantly greater in AIP and BP samples, compared with control samples. Absorbance per microgram of protein did not differ between BP and AIP samples. Blood concentrations of 3MEIN were significantly greater in cattle with AIP, compared with healthy cattle or cattle with BP. Odds of an animal with AIP being a heifer was 3.1 times greater than the odds of that animal being a steer. CONCLUSIONS AND CLINICAL RELEVANCE: Increased pulmonary production of 3MEIN may be an important etiologic factor in feedlot-associated AIP.

Putative uremic encephalopathy in horses: five cases (1978-1998).

OBJECTIVE: To determine historical, physical examination, clinicopathologic, and postmortem findings in horses with putative uremic encephalopathy. Design-Retrospective study. Animals-5 horses with renal failure and neurologic disease not attributable to abnormalities in any other organ system. PROCEDURE: Medical records from 1978 to 1998 were examined for horses with renal disease and neurologic signs not attributable to primary neurologic, hepatic, or other diseases. Signalment, history, physical examination findings, clinicopathologic data, renal ultrasonographic findings, and postmortem data were reviewed. RESULTS: Of 332 horses with renal disease, 5 met selection criteria. Historical findings, physical examination findings, clinicopathologic data, ultrasonographic data, and postmortem findings were consistent with chronic renal failure. Swollen astrocytes were detected in all 4 horses examined at necropsy. CONCLUSIONS AND CLINICAL RELEVANCE: A single criterion was not determined to be pathognomonic for uremic encephalopathy in horses. Uremic encephalopathy should be considered as a differential diagnosis in horses with evidence of chronic renal failure and encephalopathic neurologic sign not attributable to other causes. Astrocyte swelling, which was common to all 4 horses examined at necropsy, may serve as a microscopic indicator of uremic encephalopathy in horses.

Effects of water sulfate concentration on performance, water intake, and carcass characteristics of feedlot steers.

Two hundred forty single-source, cross-bred steers (304 kg) were used to evaluate the effects of various water sulfate concentrations on performance, water intake, and carcass characteristics of feedlot steers. Cattle were stratified by weight and assigned within weight blocks to five water treatments. Averaged over time, actual water sulfate concentrations (+/- SEM) were 136.1 (+/- 6.3), 291.2 (+/- 15.3), 582.6 (+/- 16.9), 1,219.2 (+/- 23.7), and 2,360.4 (+/- 68.2) mg/L, respectively. Weather-related data were recorded. Increasing water sulfate concentration resulted in linear decreases in ADG (P < 0.01) and gain:feed ratio (P < 0.01) and a quadratic effect on water intake (P = 0.02) and tended to quadratically increase then decrease DMI (P = 0.13). Sulfate x period interactions were evident for DMI (P = 0.01), ADG (P < 0.01), and feed efficiency (P < 0.01). Time had quadratic effects on DMI, water intake, ADG, and feed efficiency (P < 0.01 for all models). Increasing water sulfate concentration resulted in linear decreases in final weight, hot carcass weight, and dressing percentage, a linear increase in longissimus muscle area, and a quadratic effect on fat thickness over the 12th rib and predicted yield grade (P < 0.05 for all dependent variables). Mean daily temperature explained 25.7% of the observed variation in water intake. Other factors that explained a significant (P < 0.01) amount of variation in water intake were BW, DMI, water sulfate concentration, barometric pressure, wind speed, and humidity. High water sulfate concentrations had a significant and deleterious effect on performance and carcass characteristics of feedlot steers. Increasing the sulfate concentration in water may have resulted in a functional water restriction early in the trial when ambient temperatures were greatest. However, toward the latter stages of the trial, cattle supplied higher-sulfate water had higher ADG and FE. These improvements later in the trial may represent compensatory gain associated with decreased ambient temperature and water requirements. Averaged over time, a water sulfate concentration of greater than 583 mg/L, equivalent to 0.22% of the diet, decreased feedlot performance.

Update on sulfur-related polioencephalomalacia.

Sulfur-related polioencephalomalacia is frequently associated with a high total sulfur intake by ruminants. The onset of clinical signs coincides with excessive ruminal sulfide production. Measurement of ruminal gas cap hydrogen sulfide makes it possible to identify cattle with potentially hazardous total sulfur intake. Evaluation of all potential sulfur sources is necessary to estimate total dietary sulfur concentration, which can lead to prevention strategies.

Potential for disruption of central nervous system tissue in beef cattle by different types of captive bolt stunners.

The application of pneumatic-powered air injection stunners (PPAISs), pneumatic-powered stunners (PPSs), and cartridge-fired stunners (CFSs) in commercial beef slaughter plants was evaluated to determine the extent of dissemination of central nervous system tissue. Fifteen beef slaughter plants in the western and central United States were visited to observe stunning methods and the condition of the hearts at postmortem inspection. As inspectors performed the normal opening of the hearts, the research observer evaluated the contents of the heart for the presence of clots and/or visible tissue segments in the right ventricle. In eight plants where PPAISs were used, 33% of hearts examined (n = 1,050) contained large clots in the right ventricles. In the four plants where CFSs were used, 1% of the hearts (n = 480) contained detectable clots. In three plants where the newly modified PPSs were used, 12% of the hearts (n = 450) contained detectable clots. Large segments of spinal cord were detected, collected, photographed, and confirmed histologically from two hearts in a plant that used a PPAIS. Most of the material was found in a single right ventricle and was composed of 10 to 13 cm segments of spinal cord.

An enzyme-linked immunosorbent assay for glial fibrillary acidic protein as an indicator of the presence of brain or spinal cord in meat.

The current methods to detect central nervous system (CNS) tissue in blood, lungs, or meat are cumbersome, time consuming, and costly. The objective of this study was to use glial fibrillary acidic protein (GFAP), which is restricted to the CNS, in an enzyme-linked immunosorbent assay (ELISA) for the detection of CNS tissue in blood and muscle from beef cattle. Bovine brain, cerebral cortex, spinal cord, sciatic nerve, diaphragm, blood clots, and other skeletal muscle were obtained from three animals at slaughter. The limit for detection of GFAP was approximately 1.0 ng and the standard curve was linear up to 40 ng. Tissue samples gave responses parallel to the GFAP standard, suggesting that standard and unknown samples were immunoreactively identical. No GFAP was detected in skeletal muscle (ground beef, shoulder clod, and diaphragm) and blood clots. Trace amounts (13.5 to 51 ng/mg) were present in sciatic nerve. In contrast, high levels of GFAP (55 to 220 microg/ mg) were present in spinal cord, cerebral cortex (17 microg/mg), and whole brain (9 to 55 microg/mg). In a storage study using two animals in two separate studies, immunoreactive GFAP was detectable for up to 8 days at 4 degrees C in all tissues containing neural elements. Thus, mixtures of muscle with spinal cord or brain retained almost 80% of their immunoreactivity after 8 days at 4 degrees C, while brain and spinal cord alone retained approximately 50% and 25%, respectively, of their initial activities. In a repeat experiment, 80 to 100% of the initial activity was retained in these tissues after 8 days at 4 degrees C. The results of the current study demonstrate that the GFAP ELISA provides a valid and repeatable method to detect CNS tissue contamination in meat.

Association of excess sulfur intake and an increase in hydrogen sulfide concentrations in the ruminal gas cap of recently weaned beef calves with polioencephalomalacia.

During a 2-week period, 16 of 150 recently weaned calves developed signs of polioencephalomalacia (PEM). One calf was examined and treated at our veterinary teaching hospital and a necropsy was performed on a calf that died. During the peak of the outbreak, clinicians visited the ranch. Ruminal hydrogen sulfide (H2S) and blood thiamine concentrations were measured in 10 clinically normal penmates of PEM-affected calves. Ruminal H2S concentrations were high (> 0.695 mg/L [> 500 ppm]) in all cattle (mean, 12.19 mg/L [8,770 ppm]). All blood thiamine values were within the reference range. Within 12 hours after measurement of blood thiamine concentrations, 2 of the calves from which samples were obtained developed clinical signs of PEM. Dietary analysis revealed an estimated sulfur intake of 0.9% per calf on a dry-matter basis. Hay contributed most of this sulfur. In the investigation reported here, an outbreak of PEM was associated with high ruminal H2S concentrations and excess sulfur intake without evidence of thiamine deficiency.

Polioencephalomalacia.

Polioencephalomalacia (PEM) is a neuropathologic condition of ruminants that can be induced by a variety of neural metabolic disruptions. These include altered thiamine status, water deprivation-sodium ion toxicosis, lead poisoning, and high sulfur intake. Investigations of sulfur-related PEM have demonstrated that the onset of the clinical signs coincides with excessive ruminal sulfide production. A number of ruminal factors could modulate the production and absorption of ruminal sulfide. The development of a convenient method to estimate ruminal gas cap H2S has made it possible to identify cattle with high levels of ruminal H2S and evaluate their risk of developing PEM.

Idiopathic eosinophilic enteritis in four cattle.

Four cattle had chronic diarrhea and had lost weight but were not anorectic or dehydrated. A diagnosis of eosinophillic enteritis was made on the basis of a combination of clinical and histopathologic findings and exclusion of other diseases. Eosinophilia and hypoalbuminemia were not detected in CBC analyses. Gross and histologic examination of the small intestine revealed edema, diffuse eosinophilic inflammation (> 20 eosinophils/HPF [400 x]), and regional lymphadenopathy. A causative agent was not identified in any of the cattle. Treatment consisting of a 1-month course of systemically administered corticosteroids was attempted in 2 cattle and was apparently successful in both. Diagnostic criteria for eosinophilic enteritis have not been established for cattle and, therefore, the diagnosis was subjective. This disease resembled paratuberculosis (Johne's disease) in clinical appearance but was distinguished on the basis of a lack of a contagious etiologic agent and differences in histologic appearance.

Evaluation of ruminal sulfide concentrations and seasonal outbreaks of polioencephalomalacia in beef cattle in a feedlot.

OBJECTIVES: To measure concentrations of thiamine in blood and sulfide in ruminal fluid in cattle with polioencephalomalacia (PEM) and to evaluate temporal associations between PEM and risk factors. DESIGN: Epidemiologic analysis. SAMPLE POPULATION: 14 steers with acute signs of PEM, 26 clinically normal steers and records of all cattle in a feedlot for the past 6 years. PROCEDURES: Concentrations of thiamine in blood and sulfide in ruminal fluid were measured. Values were compared between healthy steers that had been in the feedlot for 3 weeks or 2 months. Records were used to estimate the incidence of PEM and the time when cattle were at greatest risk of developing PEM. RESULTS: Thiamine concentrations in steers with PEM were within reference ranges. Healthy steers had significantly greater sulfide concentrations 3 weeks after entering the feedlot, when the incidence of PEM was greatest, than 2 months after entering the feedlot, when risk of developing PEM was low. Thiamine concentrations were within reference ranges at these times. Annually recurrent outbreaks of PEM during the summer began after initiating use of a water well containing a high content of sulfate. CLINICAL IMPLICATIONS: Excessive ruminal sulfide production is an important factor in the pathogenesis of PEM, without concurrent thiamine deficiency. Most cases of PEM developed between 15 and 30 days after introduction to a high-sulfur diet. When water is an important source of dietary sulfur, risk of PEM may increase during hot weather.

In vivo indicators of pathologic ruminal sulfide production in steers with diet-induced polioencephalomalacia.

Two groups of 3 120-160-kg Holstein steers were fed a diet high in carbohydrate and low in long fiber and either with or without added sodium sulfate. Prior to and during the course of feeding the experimental diet, the concentrations of rumen hydrogen sulfide gas and rumen fluid sulfide were determined by a simple sulfide detector tube method and by sulfide-selective electrode, respectively. Other measurements included rumen fluid pH, blood creatine kinase, and blood sulfhemoglobin. Two of the 3 steers fed the high-sulfate diet developed signs and lesions of polioencephalomalacia. Clinical signs included episodic ataxia and blunted or absent menace reaction. Increased ruminal H2S gas concentrations occurred in all 3 steers consuming the diet with added sulfate. The onset of clinical signs coincided with the onset of elevated H2S concentrations. These increases were 40-60 times the values measured in the steers consuming the diet without added sulfate. In contrast, increases in rumen fluid sulfide concentrations usually rose to 4 times that of control steers. The steers fed an identical diet but without added sulfate exhibited no signs or lesions of polioencephalomalacia and no elevations of sulfide in rumen gas or fluid. All steers had a modest decrease in rumen fluid pH associated with the transition to the concentrate diet. No significant changes were observed in any of the blood measurements of any of the steers. An additional pair of steers was fed the experimental diet with or without added sulfate to compare the ruminal H2S gas concentrations estimated by H2S detector tubes with those estimated by a different method of analysis utilizing charcoal trapping of H2S, conversion to sulfate, and measurement of the sulfate. Both methods yielded comparable estimates of H2S concentration. Overall, these data indicate that changes in rumen gas cap H2S concentrations are larger than changes in rumen fluid sulfide concentration and the estimation of rumen gas cap H2S concentration may be a practical approach to detecting pathologic increases in ruminal H2S gas. This simple, rapid, minimally invasive method should be useful for estimating the H2S content of ruminal gas under field conditions.

Identity and interactions of rumen microbes associated with dietary sulfate-induced polioencephalomalacia in cattle.

To study their role in sulfate reduction, anaerobic bacteria were cultured from rumen fluid samples of cattle fed high-carbohydrate, short-fiber diets with and without added sulfate. The steers fed the diet with added sulfate developed polioencephalomalacia. Microbiological methods included colony type profiles, molybdate sensitivity, presence of desulfoviridin, sulfate reduction rates of pure and mixed cultures, and incubation time effects on sulfate reduction. Colony-type profiles indicated decreased diversity, but no relative change in numbers of sulfate-reducing bacteria in rumen fluid from cattle fed diets with and without added sulfate. Thirteen bacteria] isolates were selected for further study on the basis of colony type, sulfate-reducing activity, and growth in lactate, sulfate, and yeast extract media. Seven of the isolates had Desulfovibrio-like characteristics (ie, they were gram-negative, motile rods that reduced sulfate, were inhibited by molybdate, and contained the pigment desulfoviridin). The remaining 6 isolates were gram-negative, nonmotile rods. Four of these released sulfide from cysteine, and 2 generated only limited amounts of sulfide from sulfate or cysteine. The 7 sulfate reducing isolates generated sulfide in rumen fluid broth medium at greater rates than those observed in fresh rumen fluid. Sulfate reduction Could be sustained in cultures for prolonged incubation times if the gas phase containing hydrogen sulfide was replaced at frequent intervals. Variations in the amount of sulfate reduced by the pure cultures were most pronounced at short incubation times. Sulfate reduction was not inhibited in mixed cultures of sulfate-reducing and nonsulfate-reducing bacteria.

Ruminal microbial alterations associated with sulfide generation in steers with dietary sulfate-induced polioencephalomalacia.

Holstein steers were fed carbohydrate-rich, short-fiber basal diets with and without added sodium sulfate. Steers fed the high-sulfate diet developed the CNS disorder polioencephalomalacia (PEM). The onset of signs of PEM was associated with increased sulfide concentration in the rumen fluid. Over the course of the disease, anaerobic rumen bacteria were enumerated in roll tubes by use of the Hungate method Lo determine the effect of dietary sulfate on sulfate-reducing bacterial numbers. Media used included a general type for total counts and sulfate containing media with and without cysteine to assess sulfate-reducing bacteria. Changes in total and sulfate reducing bacterial numbers attributable to dietary sulfate content were not observed. The capacity to generate hydrogen sulfide from sulfate in fresh rumen fluid in vitro was substantially increased only after steers had been fed the high sulfate diet for 10 to 12 days, which coincided with the onset of signs of PEM. The low capacity for hydrogen sulfide production of rumen fluid taken at earlier times in the feeding period suggests that rumen microorganisms must adapt to higher dietary sulfate content before they are capable of generating potentially toxic concentrations of sulfide.

High-performance liquid chromatography detection of sulfide in tissues from sulfide-treated mice.

The biological and forensic use of ion-interaction reversed-phase high-performance liquid chromatography for the determination of hydrogen sulfide-derived methylene blue is evaluated by measuring the sulfide content in tissues from sulfide-treated mice. Various preparative conditions were examined. The determinations of background levels of sulfide from brain, liver and kidney were compared to sulfide levels from mice exposed to 60 micrograms g-1 sodium hydrosulfide. At the time of death, significant increases above background sulfide levels were measured for all three biological tissues. To evaluate its forensic potential, we used this sulfide detection methodology to evaluate comparatively the sulfide levels from fresh and frozen samples of brain, liver and kidney. The stability of sulfide levels obtained from frozen brain makes this tissue the most reliable tissue for forensic evaluation. Samples of brain, liver and kidney obtained within 24 h of death by sulfide intoxication had demonstrable elevations in sulfide concentration.

Parvovirus-associated syndrome (Aleutian disease) in two ferrets.

There is a paucity of information regarding natural Aleutian disease, caused by a parvovirus in ferrets. With the increasing popularity of ferrets as household pets and laboratory animals, and with the advent of a USDA-approved rabies vaccine, the occurrence and the etiopathogenesis of naturally acquired diseases in ferrets needs to be documented. We present the clinical and laboratory findings associated with Aleutian disease in 2 domestic ferrets, one with the chronic wasting form of the disease and one with the central nervous system form.