Discolored Urine in Horses and Foals.

This article describes the most common causes of urine discoloration. The review includes a description of the most common disorders causing hematuria, highlighting clinical presentation, treatments, and pathophysiology. Causes of hemoglobinuria and myoglobinuria together with their mechanisms of renal injury are also reviewed.

Major Histocompatibility Complex Dmα/ß Genes and Their Expressional Diversity in Healthy and Diseased Water Buffalo Bubalus bubalis.

BACKGROUND/AIMS: The major histocompatibility complex (MHC) categorized into three (I, II and III) classes elicits the immunogenic response by presenting exogenous peptides to T cells. The MHC-II DM is composed of DMα and DMß, two polypeptide chains, both are encoded by separate MHC genes involved in antigen processing and presentation. Despite the acknowledged role of MHC complex in humans, the literature is silent on the organization and expression of these genes in water buffalo Bubalus bubalis, an agriculturally important animal species. METHODS: We deduced the full-length mRNA sequences of DMα and DMß genes, localized them onto the chromosome 2, assessed their copy number per haploid genome and studied tissue and disease specific expression. RESULTS: The Real Time PCR showed higher expression of both the genes and their seven interacting partners in spleen, gonads and spermatozoa. Significantly, upregulation of DMα and DMß genes and their interacting partners were detected in diseased group of buffaloes as compared to that in healthy ones. CONCLUSION: The upregulation of Bubalus bubalis (BuLA)-DMα and DMß genes and their interacting partners reflect their role in regulating immune responses towards the amelioration of diseases. Work on this line would enhance our understanding on the overall roles of MHC locus, allowing development of possible therapeutic treatment strategies.

Pathology of Naturally Occurring Bacillary Hemoglobinuria in Cattle.

Clostridium haemolyticum causes bacillary hemoglobinuria (BH), an infectious and usually fatal disease that occurs mostly in cattle, which is clinically characterized by jaundice, hemoglobinuria, and anemia. The trematode Fasciola hepatica has been commonly reported as the main predisposing factor that triggers this condition. The authors evaluated 20 naturally occurring cases of bovine BH to characterize the pathology and pathogenesis of the disease. Grossly, the most consistent finding was a large, frequently single focus of necrosis surrounded by a red to purple halo, observed most frequently on the parietal surface of the right and left hepatic lobes. Other findings were jaundice, dark-brown discoloration of kidneys, and red urine in the urinary bladder. Microscopically, characteristic lesions were locally extensive, necrotizing hepatitis with thrombosis and numerous intralesional Gram-positive rod-shaped bacteria, and acute renal tubular necrosis. By immunohistochemistry, many hepatocytes outside the necrotic focus in the liver were positive for activated caspase 3, suggesting that those cells were undergoing apoptosis. Ultrastructural evaluation revealed hepatocyte necrosis, hemolysis, and clumps of vegetative and sporulating bacilli within the liver. Polymerase chain reaction for the C. haemolyticum beta toxin gene was positive in randomly selected liver samples. No gross or microscopic lesions indicative of fascioliasis were detected in the liver of any animal, suggesting that other yet undetermined predisposing factors were associated with these cases of BH.

Diagnostic exercise: hemolysis and sudden death in lambs.

Within a 24-hour period, 7 out of 200 three- to four-week-old pastured Katahdin lambs died after showing clinical signs of hemoglobinuria, red-tinged feces, weakness, and recumbency. One of the lambs that was examined clinically before natural death also had abdominal pain, trembling, tachycardia, and severe anemia with a packed cell volume of 4%. Pathologic findings included icterus, hemoglobinuric nephrosis, dark red urine, pulmonary edema, hydrothorax, splenomegaly, and acute centrilobular to midzonal hepatocellular degeneration and necrosis with cholestasis. The differential diagnoses and diagnostic workup to achieve the diagnosis are briefly discussed.

A case of haemoglobinuria in a cat after near-drowning.

The aim of this study is to describe a case of haemoglobinuria in a cat after near-drowning. A 6-year-old male neutered domestic short hair cat weighing 6.5 kg with a pre-existing seizure disorder presented to an emergency hospital after near-drowning in a swimming pool during a seizure episode. On presentation, the patient was obtunded, dyspnoeic, bradycardic and hypothermic. Imaging revealed evidence of severe bilateral pulmonary infiltrates. Treatment with intravenous diazepam, amoxicillin, fluid therapy, active warming and oxygen therapy was administered. The cat developed haemoglobinuria approximately 6 h after nearly drowning. Despite improvements in mentation, pulse quality and heart rate, respiratory compromise and poor oxygen saturation persisted, prompting euthanasia approximately 10 h after admission. To the author's knowledge, this is the first reported clinical case of haemoglobinuria following near-drowning in veterinary medicine.

Discolored urine in sheep and goats: Clinical, etiological, hematobiochemical, sonographic and postmortem findings.

Background: Bloody urine is classified in farm animals as hematuria, hemoglobinuria, and myoglobinuria. In small ruminants, discolored urine is reported due to several etiologies which is sometimes fatal. Of these causes are babesiosis, bacillary hemoglobinuria, copper toxicity, and hypophosphatemia. Aim: This study was designed to investigate the clinical, etiological, hematobiochemical, ultrasonographic, and pathological findings in rams and bucks with red urine syndrome. Methods: Eighteen male animals (nine rams and nine bucks) of 6 months to 3 years were examined. Parallel, 10 healthy controls were used. They were admitted due to red urine, voiding of only urine drops, straining during the act of urination, grunting during urination, ventral abdominal edema, and abdominal distension. The duration of the disease ranged from 2 to 30 days. A history of chronic copper toxicosis was informed in two bucks and a ram. Two blood samples were collected from diseased as well as from controls in EDTA tubes (for complete blood count testing) and in plain tubes (for serum collection). Results: Hematuria was found in 11 animals (seven bucks and four rams) while hemoglobinuria was detected in seven animals (five bucks and two rams). Sonographic findings in diseased animals included ruptured urinary bladder in 3, ruptured urethra in 5, penile calculi, uroperitoneum in 6, distended urinary bladder in 7, hydronephrosis in 5, echogenic deposits in the bladder in 3, and ventral urine accumulation in four animals. Laboratory evaluation of a Geimsa-stained blood smear confirmed the infection with Babesia in three bucks and a ram. Hemolytic anemia was marked in two bucks and a ram due to chronic copper toxicity. Biochemical abnormalities included hypoalbuminemia, hyperglobulinemia, increased blood urea nitrogen and creatinine concentration, and hyperglycemia. Postmortem examination was carried out on six animals (four rams and two bucks). Conclusion: Discolored urine in rams and bucks in this study resulted from hematuria due to urinary calculi and pelvic abscessation or from hemoglobinuria due to Babesia infection or due to copper toxicity. Hemolytic anemia was the outstanding hematological finding and hypoalbuminemia, hyperglobulinemia, increased blood urea nitrogen (BUN) and creatinine, and hyperglycemia were the characteristic biochemical findings. Sonography of the urinary tract was very helpful in assessing the renal parenchyma, urinary bladder, and abdominal cavity for the verification of urolithiasis, hydronephrosis, intact or ruptured urinary bladder, uroperitoneum, and perforated urethra.

Spatial and seasonal analysis and geovisualization of Fasciola hepatica-free bovine bacillary hemoglobinuria outbreaks in eastern Uruguay, 1999-2019.

Bovine bacillary hemoglobinuria (BBH) produced by Clostridium novyi type D, is an endemic, highly fatal disease of cattle in the temperate grassland region of eastern Uruguay. A previous study showed that in this region, BBH is not associated with Fasciola hepatica or any other known focal-ischemic liver injury, so the reasons for its high incidence remains undetermined. The objective of this study was to analyze data from 45 Fasciola hepatica-free BBH outbreaks (1999-2019) in order to find common animal, seasonal and/or geographical risk factors, which may explain the occurrence of the epizootics. Fisher's goodness-of-fit testing showed a significantly higher case proportion of adult cows (N = 368, 80.5%) and lower of calves (N =8, 1.8%), as compared to the expected proportions of the censused population in the study area and historical submissions computed from the laboratory database (Chi-Sq = 346.2 and 174.8, df = 7, P < 0.00). Time series decomposition showed a bi-seasonal pattern, with a larger peak in spring and early summer (October to January) and a smaller increase in autumn (March-May). The lowest seasonal indices were on mid-summer (February) and winter (June-September). A combination of spatial statistics was used to assess the different spatial features of the disease and consistency of the findings. Global spatial autocorrelation showed BBH was significantly clustered (Moran's I = 0.407, P < 0.001). Both smoothed Anselin's Local Indicator of Spatial Autocorrelation and Kulldorff's spatial scan Poisson and Bernoulli models, detected roughly the same high-risk areas in the southeastern part of the Merin Lagoon basin, with the most likely cluster centered in the large wetland biosphere reserve "Eastern Wetlands and Coastal Strip" (RR = 9.12, P < 0.001). Outbreaks were georeferenced (latitude, longitude) and thematic dot-mapping geovisualization in Google Earth™ showed that the results were robust and truly geographic in nature. Most outbreaks (40/45, 88.8%) occurred on wetlands areas and large river valleys, characterized by poorly drained and frequently flooded soils, indicating that moisture-laden soils are the natural habitat of C. novyi type D. Grasslands in these endemic areas support rapid fattening of cattle during spring-summer, and somewhat less in autumn, in almost exact correspondence with BBH peaks, suggesting a close causal association in high-risk areas. Risk is significantly higher in adult cows probably because the spore content in the liver is highest in this category. The altered lipid metabolism and lipotoxicity in the liver may be the precipitating factor for spore germination and epizootic occurrence.

Changes to blood parameters after postparturient hemoglobinuria in 11 Holstein-Friesian cows.

This study reviewed a case series of 11 Holstein-Friesian (HF) cows with postpartum hemoglobinuria (PPH) from one dairy herd. The first clinical signs of PPH appeared in the animals during the second or third lactation, between 21 and 30 days after calving. The clinical signs, including depression, diminished appetite, a dark red to brown color in the urine, pale mucous membranes, and a decrease in milk yields were observed in these 11 animals. Three of the cows developed jaundice of the mucous membranes and five had dry, parched feces. PPH was confirmed on laboratory test results of blood and urine samples. Anemia, serum hypophosphatemia (Pi = 0.79 mmoL/L), and increased liver function analytes (total bilirubin, total protein, and urea concentrations) were observed in all animals. Animals were treated with intravenous phosphorus supplementations for the first 2 days after clinical signs were noted, and then oral supplementations were administered. After the clinical signs resolved and the treatments were discontinued, the animals still had mild anemia; however, the phosphorus concentration increased to 1.40 mmoL/L. Gamma-glutamyltransferase activity increased compared with activities measured before treatments and total bilirubin concentrations decreased slightly; however, the concentrations were still more than twice the upper limit of the normal RI. These animals were diagnosed with liver damage that had developed over the course of PPH, indicating the need for the further monitoring and treatment of cows during the postparturient period, even if clinical signs are no longer present.

Exertional hemolysis and hematuria in a Labrador Retriever dog.

A 7-y-old male Labrador Retriever dog was presented because of acute onset of dark-colored urine after a hunting session the day prior. Moderate hemoglobinemia was observed, associated with transient hemoglobinuria and hematuria with no concurrent evidence of underlying urinary tract disease. The patient's clinical signs resolved within 36 h post-exercise without specific treatment. The concurrent occurrence of exertional hemolysis and hematuria in a dog is uncommon; these conditions are commonly reported separately in human athletes.

Bacillary hemoglobinuria in Japanese black cattle in Hiroshima, Japan: a case study.

Three Japanese Black cows housed with 6 other cows exhibited main clinical symptoms of severe hemoglobinuria. Hematological analyses conducted after antibiotic therapy demonstrated severe anemia, and biochemical analyses indicated both severe hemolysis and disruption of hepatic function. One of the three cows died. Based on the above analyses and observation of typical clinical symptoms, a speculative diagnosis of bacillary hemoglobinuria was made, and immediate high-dose antibiotic treatment improved the general conditions of the surviving animals. Blood samples from the other 2 cows were collected sequentially after antibacterial therapy. Clostridium haemolyticum was detected by a nested polymerase chain reaction analysis of the blood samples. The cows were diagnosed with the second recorded occurrence of bacillary hemoglobinuria in Japan.

Copper Toxicity in Horses: Does it Exist?

Copper toxicity is thought to be a rare condition in horses. However, the number of cases diagnosed in Brazil is growing. This article aims to describe cases of copper toxicity involving horses from different geographic locations and discuss findings of physical examinations, differential diagnoses and potential causes. Five cases referred from 4 different properties where at least 15 other horses were affected were described. Hemolytic anemia and hemoglobinuria, presence of Heinz bodies and elevated aspartate aminotransferase and gamaglutamil transferase levels were detected in all cases. The diagnosis was based on clinical history and signs, laboratory tests results, copper level determination in feed and/or soil and histopathological findings. Two horses progressed to acute death; remaining horses responded to clinical management with or without blood transfusion, depending on disease severity. However, one of these horses, after several returns to the veterinary hospital, was euthanized due to complications. One horse was treated with ammonium tetrathiomolybdate. Two horses had several recurring episodes over the course of several months, an uncommon presentation in ruminants suffering from copper toxicity. Excess copper was associated with soil fertilization with poultry litter or treatment of previous or neighbor crops with copper-containing products. It can be concluded that copper toxicity does occur in horses and may arise from several sources and/or be associated with predisposing dietary factors. Given the growing number of cases, the condition should be included in the differential diagnosis list and proper preventive dietary and pasture fertilization measures adopted.

Pathobiology and diagnosis of clostridial hepatitis in animals.

Clostridia can cause hepatic damage in domestic livestock, and wild and laboratory animals. Clostridium novyi type B causes infectious necrotic hepatitis (INH) in sheep and less frequently in other species. Spores of C. novyi type B can be present in soil; after ingestion, they reach the liver via portal circulation where they persist in phagocytic cells. Following liver damage, frequently caused by migrating parasites, local anaerobic conditions allow germination of the clostridial spores and production of toxins. C. novyi type B alpha toxin causes necrotizing hepatitis and extensive edema, congestion, and hemorrhage in multiple organs. Clostridium haemolyticum causes bacillary hemoglobinuria (BH) in cattle, sheep, and rarely, horses. Beta toxin is the main virulence factor of C. haemolyticum, causing hepatic necrosis and hemolysis. Clostridium piliforme, the causal agent of Tyzzer disease (TD), is the only gram-negative and obligate intracellular pathogenic clostridia. TD occurs in multiple species, but it is more frequent in foals, lagomorphs, and laboratory animals. The mode of transmission is fecal-oral, with ingestion of spores from a fecal-contaminated environment. In affected animals, C. piliforme proliferates in the intestinal mucosa, resulting in necrosis, and then disseminates to the liver and other organs. Virulence factors for this microorganism have not been identified, to date. Given the peracute or acute nature of clostridial hepatitis in animals, treatment is rarely effective. However, INH and BH can be prevented, and should be controlled by vaccination and control of liver flukes. To date, no vaccine is available to prevent TD.

Copper toxicosis with hemolysis and hemoglobinuric nephrosis in three adult Boer goats.

Acute and, particularly, chronic copper exposures, along with defects in hepatic copper metabolism, altered excretion of copper, and/or nutritional imbalances between copper and other trace elements, can lead to hepatic accumulation of copper and primary copper toxicosis. There is interspecies variation in susceptibility to copper toxicosis, with sheep being the species most likely to develop this condition. Adult dairy goats and Boer crosses are generally considered resistant to chronic copper toxicosis, especially the hemolytic stage of this disease. The current report is rather unusual in that it describes instances of naturally occurring copper toxicosis with hemolysis and hemoglobinuric nephrosis in 3 adult Boer goats. In 2 of these goats, a possible source of excessive dietary copper was investigated but not definitively identified. In the third goat, the etiologic factors associated with the copper toxicosis were not determined. It appears that mature Boer goats are susceptible to the hemolytic stage of chronic copper toxicosis, which was not observed in a recent, large-scale copper intoxication involving lactating dairy goats. Copper analyses on both liver and kidney samples were necessary to confirm the diagnosis of copper toxicosis in all 3 goats. All feedstuffs associated with instances of copper toxicosis should be analyzed for iron, molybdenum, sulphur, and zinc as well as copper to determine what nutritional factors are contributing to the pathogenesis of this disease. Consideration also should be given to the ingestion of hepatotoxic plants and other toxic exposures, which could predispose an animal to secondary chronic copper toxicosis.

Successful treatment of bacillary hemoglobinuria in Japanese Black cows.

Four pasture-fed Japanese Black cows showed the main clinical symptoms of severe hemoglobinuria at different periods between 2003 and 2007. Hematological analyses at the first consultation revealed severe anemia, and biochemical analyses indicated both severe hemolysis and disruption of hepatic function. Although the first 2 patients died, the hemoglobinuria and general condition of the remaining 2 cows, who were immediately initiated on large doses of antibiotics, improved within 3 days. Clostridium haemolyticum was detected by polymerase chain reaction (PCR) analysis of the blood sample of 1 of the infected cows. Anti-fascioliasis medicine is administered, and since then, no case of hemoglobinuria has been observed. The cows were diagnosed with bacillary hemoglobinuria, and they represent the first few cases in Japan.

Recovery with a regular dose of antibiotics from bacillary hemoglobinuria in a Holstein cow.

One Holstein cow housed with 21 other cows exhibited clinical signs of pyrexia, anorexia and diarrhea along with severe hemoglobinuria. Hematological and biochemical analyses conducted before and after antibiotic therapy indicated severe hemolytic anemia and disruption of hepatic function. A general improvement in conditions was observed after an 11-day program of treatment comprising a regular dose of antibiotics and prescribed supportive therapies. A tentative diagnosis of bacillary hemoglobinuria was made based on the clinical and clinico-pathologic features on day 7. A molecular diagnosis was made by a PCR amplification of the flagellin gene of Clostridium haemolyticum using DNA extracted from the whole blood. The cow was diagnosed with the first recorded occurrence of bacillary hemoglobinuria of Holstein cattle in Japan.

Acute post-parturient haemoglobinuria in dairy cows and phosphorus status.

During the conduct of an experiment designed to examine the nutritional management of dairy cows in late pregnancy, four cows out of 72 suffered from acute haemoglobinuria two to four weeks after calving. Thirty-six thin and 36 fat cows were individually fed one of three diets based on a total mixed ration with different energy or protein concentrations during the last 3 to 4 weeks before expected calving date. After calving, cows grazed pasture and were offered 6 kg dry matter of pelleted concentrates daily. The P concentrations of the feeds offered suggested that the cows' diets were marginally deficient in P relative to requirements. Plasma P concentrations were significantly (P < 0.05) lower in fat cows than in thin cows during the first 6 weeks of lactation (0.87 versus 1.12 mmol/L), but precalving diet had no effect (P > 0.05). Concentrations of plasma inorganic P of the four fat cows that developed acute haemoglobinuria were less than 0.3 mmol/L. However, plasma P concentrations in another 12 cows, none of which displayed overt symptoms, declined to similar levels. It appeared that inadequate dietary P may have predisposed cows to acute haemoglobinuria, but the precipitating cause was not readily obvious.

Variation of serum inorganic phosphorus and association with haemoglobinuria and osteomalacia in female water buffaloes in Pakistan.

Data from an animal health service in the Punjab of Pakistan showed that 39 adult female buffaloes with haemoglobinuria were 21 times more likely to have serum inorganic phosphorus (serum P) levels < 0.97 mmol/l than 24 controls sampled during the period of case occurrence (December 1984-March 1985). Age > 7 years or early lactation (1-60 days post partum) were unrelated to the disease. Similarly, symptoms of osteomalacia in 19 multiparous buffaloes were associated with low P (OR = 14.3) but not with age. Subsequently, a serum survey was carried out from February 1985 to July 1987 to investigate serum P variations with season and host factors. Data from 139 farms (426 adult female buffaloes, 468 lactations) indicated strong farm and seasonal effects on serum P. Serum P declined during the study period and was lowest during December-March 1985/1986 and again 1986/1987. Calving season, parity > 1, high pregnancy > 6 months, or daily milk production were not related to serum P in the final model. Seasonal effects were interpreted as soil borne and related to feed changes from maize to berseem in December.

Hemoglobinuric nephrosis in a rhea (Rhea americana).

An eighteen-month-old female rhea (Rhea americana) was presented dead for necropsy. The owner reported having observed blood in the droppings. Gross examination revealed a rhea in good body condition with a copious amount of frank blood in the cloaca. Large masses of matted fescue grass (Festuca spp.) distended the ventriculus and jejunum. No hemorrhage was evident in the digestive tract, which was otherwise void of ingesta. The kidneys were dark brown in color. Renal histopathology revealed a severe accumulation of eosinophilic pigment in the tubular epithelial cells of the proximal convoluted tubules, with tubular nephrosis and eosinophilic casts in the collecting tubules. It was concluded that the renal pigment was hemoglobin and not myoglobin, based on lack of evidence of muscle injury and the severe erythrophagocytosis evident in hepatic macrophages. The renal pigment also stained positive with the hemoglobin-specific Okajima stain. Hemoglobinuric nephrosis has not previously been reported in an avian species.

The experimental infection of calves with a British leptospire of the Pomona serogroup.

Two calves were infected with a British leptospire of the Pomona serogroup. Both showed symptoms of anorexia and fever and one had marked haemoglobinurea and leptospiruria. The serum antibody titres rose rapidly in both calves.

Bovine post-parturient haemoglobinuria: effect of inorganic phosphate on red cell metabolism.

Washed red blood cells from normal cows were incubated as 10 and 20 per cent suspensions in media containing 0, 2.5 and 25 mM phosphate. The results showed that the rate of glycolysis was dependent on the inorganic phosphate concentration. In the absence of phosphate, the consumption of glucose and the production of 2,3-diphosphoglycerate, lactate and adenosine 5'-triphosphate (ATP) were decreased. Incubation without added phosphate also greatly increased the production of fructose-1,6-diphosphate, glyceraldehyde-3-phosphate and dihydroxyacetone phosphate. Moderate hypophosphataemia was induced in two pre-ruminant calves. Washed red cells from the blood of these animals showed a depletion of ATP when compared with red cells from a control calf. The results indicate that phosphorus deficiency, leading to hypophosphataemia, may be a mechanism of post parturient and related syndromes of haemoglobinuria by decreasing red cell glycolysis and resultant ATP synthesis. Subnormal concentrations of ATP would predispose red cells to altered structure and function, a loss of normal deformability, and an increase in fragility and haemolysis with resultant haemoglobinuria.