Bacillary Hemoglobinuria Research Articles

Bacillary hemoglobinuria in beef cattle infected with Fascioloides magna in Missouri.

Bacillary hemoglobinuria (BH) is an infectious disease, mostly affecting cattle, caused by Clostridium haemolyticum (C. novyi type D), with acute hepatic necrosis and intravascular hemolysis. Cattle are typically predisposed to BH by liver injury caused by Fasciola hepatica, although cases have been reported in cattle without evidence of this parasite. Here we describe a cluster of 14 BH cases from 7 counties in north-central to central Missouri submitted to a veterinary diagnostic laboratory between December 2020 and April 2023. Postmortem examination in all cases revealed hemoglobinuria and acute hepatic necrosis with large numbers of gram-positive bacilli with terminal-to-subterminal spores. Flukes, fluke ova, and/or fluke pigment consistent with Fascioloides magna were identified in 12 of 14 cases. Sequences of the nuclear ribosomal internal transcribed spacer 1 (ITS1) from one fluke had 100% identity to F. magna. C. novyi was detected by fluorescent antibody testing of liver impression smears (11 of 12 cases) and by immunohistochemistry of liver sections (7 of 7 cases). PCR on formalin-fixed, paraffin-embedded tissues amplified the C. haemolyticum beta toxin gene in each of the 7 cases tested. To our knowledge, a confirmed cluster of BH associated with F. magna has not been reported previously in cattle.

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

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. This study was designed to investigate the clinical, etiological, hematobiochemical, ultrasonographic, and pathological findings in rams and bucks with red urine syndrome. 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). 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). 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.

Bacillary hemoglobinuria in a cow

Bacillary hemoglobinuria in a cow

Fist description of bacillary hemoglobinuria in a cattle in Minas Gerais, Brazil

Bacillary hemoglobinuria (BH) is a histotoxic infection caused by Clostridium haemolyticum that affects mostly cattle parasitized with trematodes. The aim of present study was to describe an atypical case of BH in a cow without access to flooded areas and without parasitism by Fasciola hepatica. After eight days of sternal decubitus and apathy, a Nellore cow was euthanized and necropsied. During postmortem examination, mild jaundice, black urine and multifocal lesions in the liver were observed. Histopathology revealed multifocal coagulation foci of necrosis in liver and basophilic bacillary structures, which were confirmed as C. haemolyticum by bacterial isolation, PCR and sequencing techniques. This is the first description of BH in cattle in Minas Gerais state, Brazil and highlight the need for of inclusion of BH as differential diagnosis even in animals not parasitized by trematodes.

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.

First report of bacillary hemoglobinuria in buffaloes / Primeiro relato de hemoglobinúria bacilar em búfalos

We herein describe a typical episode of bacillary hemoglobinuria in buffalloes, triggered by liver fluke invasion proved by pathognomonic macro and micro lesions of fasciolosis in the biliary ducts and the nephrosis and hepatic focal necrosis characterizing the toxic-infectious process caused by Clostridium haemolyticum. This report is quite similar to another one described by AHOURAI et al. (1990) in cattle following liver fluke invasion and C. haemolyticum isolation and we both agree with the fact that foci of ischemic of toxic necrosis serve as a focus in which clostridial spores might vegetate and cause the disease by producing toxins. Vaccination with bacterin or toxoids is the most effective measure for preventing these histotoxic clostridial diseases since the endospores of the microorganism persist in the environment. Also, as the process is triggered by fluke liver migration, the prevention should cover the control of this trematode as well. Although the pathogenicity of bacillary hemoglobinuria is well known, there are only a few reports currently available, and to our knowledge this is the first case of the disease described in buffaloes.

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.

Clostridial disease in cattle

Clostridial disease affects cattle across the UK throughout the year but particularly in the grazing season. All clostridial disease occurs when conditions in the muscles or organs become anaerobic allowing for bacterial proliferation and toxin production. The most common pathogenic clostridium is Clostridium chauvoei which causes blackleg and has both skeletal and cardiac forms. Clostridium perfringens is the second most commonly identified clostridia with type A causing enterotoxaemia and type D causing pulpy kidney. Clostridium novyi is the third most common clostridia — type B causes black disease and type D causes bacillary haemoglobinuria. Cattle are most at risk during the grazing season meaning that control and prevention of disease should be particularly targeted at this time of year. The cost of vaccination is so low compared with the cost of losing even one animal that an appropriate protocol should be implemented on all farms.

Clostridial diseases diagnosed in cattle from the South of Rio Grande do Sul, Brazil. A forty-year survey (1978-2018) and a brief review of the literature

ABSTRACT: Clostridial diseases are important causes of livestock losses in the southern Rio Grande do Sul. Since 1978 annual surveys conducted at the “Laboratório Regional de Diagnóstico” of the “Universidade Federal de Pelotas” (LRD-UFPel) have shown that clostridial diseases represent 10.40% of the bacterial diseases diagnosed in cattle and 1.65% of all diseases diagnosis in cattle over a 40-year period. The purpose of this study is to review the clinical, epidemiological and pathological aspects of the clostridial diseases diagnosed in cattle from January 1978 to December 2018 at the LRD-UFPel in the hopes that it will constitute a useful guide for field veterinary practitioners and interested farmers. We assessed and review the necropsy protocols of 6,736 cattle; these necropsies were performed either by LRD-UFPel faculty or by field veterinary practitioners; 111 outbreaks (1.65%) were diagnosed as clostridial disease, distributed as follows: 35 outbreaks of tetanus, 34 of blackleg, 23 of bacillary hemoglobinuria, 11 of malignant edema (gas gangrene), and eight of botulism. Approximately 904, from a total of 42,480 cattle at risk, died in these outbreaks.

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.

The Use of Next-Generation Sequencing in the Identification of a Fastidious Pathogen: A Lesson From a Clinical Setup.

Clostridium haemolyticum is the causal agent of bacillary hemoglobinuria in cattle, goat, sheep, and ruminants. In this study, we report the first recorded human-infecting C. haemolyticum strain collected from an 18-year-old woman diagnosed with acute lymphoblastic leukemia. After failure of traditional techniques, only next-generation sequencing (NGS) technology in combination with bioinformatics, phylogenetic, and pathogenomics analyses revealed that our King Faisal Specialist Hospital and Research Center (KFSHRC) bacterial isolate belongs to C. haemolyticum species. KFSHRC isolate is composed of 1 chromosome and 4 plasmids. The total genome size is estimated to be 2.7 Mbp with a low GC content of 28.02%. Comparative pathogenomics analysis showed that C. haemolyticum KFSHRC isolate is a potential virulent pathogenic bacterium as it possesses the virulence factors necessary to establish an infection, acquire essential nutrients, resist antimicrobial agents, and tolerate hostile conditions both in the human host and in its surrounding environment. These factors are included in the main chromosome in addition to novel recombination of the plasmids, and they could be the reason for the incidence of that human infection. This work demonstrated the importance of using NGS in medical microbiology for pathogen identification. It also demonstrates the importance of sequencing more microbial samples and sharing this information in public databases to facilitate the identification of pathogenic microbes with better accuracy.

Morte súbita em bovinos no Sul do Rio Grande do Sul: epidemiologia e diagnóstico

Resumo: Foi realizado um levantamento nos protocolos de necropsias de bovinos com histórico de morte súbita ou superaguda recebidos no Laboratório Regional de Diagnóstico da Faculdade de Veterinária da Universidade Federal de Pelotas (LRD/UFPel) entre 2000 e 2014. Para o estudo foram considerados os casos em que os animais tinham morrido inesperadamente sem apresentação de uma doença prévia ou sinal clínico no intervalo de 24 horas antes da observação do cadáver (Categoria 1) ou bovinos movimentados ou que sofreram algum tipo de manejo e morreram após quedas ou tremores com observação destes sinais (Categoria 2). Foram identificados 72 casos ou surtos incluídos nestas duas categorias de um total de 2.031 cadáveres/materiais de bovinos recebidos no LRD/UFPel no período, representando 3,5% do total. Os casos ocorreram em todas as épocas do ano e em 34 casos (47,2%) os bovinos afetados eram adultos, em 23 casos (31,9%) tinham entre dois e três anos e em 11 (15,3%) tinham até um ano de idade. Em quatro protocolos (5,6%) a idade não foi informada. Em 62 casos (86,1%) a forma de criação era extensiva, em sete (9,7%) a forma era semi-intensiva e em três (4,2%) a forma era intensiva. Dos 72 casos/surtos observados 52 (72,2%) foram classificados na Categoria 1 e 20 (27,8%) na Categoria 2. As enfermidades que mais causaram morte súbita ou superaguda foram: babesiose cerebral (10/72), intoxicação por organofosforados (10/72), carbúnculo hemático (7/72), hemoglobinúria bacilar (5/72) e fulguração (3/72). Dos 18 casos inconclusivos em apenas cinco foi realizada a necropsia completa e nos 15 casos negativos a Bacillus anthracis o material remetido não permitiu a pesquisa de outras enfermidades. Os resultados obtidos, permitiram concluir que as mortes súbitas na região Sul do Rio Grande do Sul são causadas por doenças, na sua grande maioria, bem conhecidas e endêmicas da região e que podem ser controladas ou evitadas por vacinação e manejo adequados. O envio de órgãos ao laboratório, quando não é possível enviar o cadáver completo, pode limitar o diagnóstico conclusivo em casos de morte súbita/ superaguda. O número expressivo de casos negativos a Bacillus anthracis ocorre em consequência de carbúnculo hemático ser uma suspeita frequente quando bovinos aparecem mortos sem a apresentação de sinais clínicos resultando no envio de material inadequado para a realização de outros diagnósticos.

Suspected Bacillary Haemoglobinuria in a Calf: A Case Report

Suspected Bacillary Haemoglobinuria in a Calf: A Case Report

Plasmidome interchange between Clostridium botulinum, Clostridium novyi and Clostridium haemolyticum converts strains of independent lineages into distinctly different pathogens.

Clostridium botulinum (group III), Clostridium novyi and Clostridium haemolyticum are well-known pathogens causing animal botulism, gas gangrene/black disease, and bacillary hemoglobinuria, respectively. A close genetic relationship exists between the species, which has resulted in the collective term C. novyi sensu lato. The pathogenic traits in these species, e.g., the botulinum neurotoxin and the novyi alpha toxin, are mainly linked to a large plasmidome consisting of plasmids and circular prophages. The plasmidome of C. novyi sensu lato has so far been poorly characterized. In this study we explored the genomic relationship of a wide range of strains of C. novyi sensu lato with a special focus on the dynamics of the plasmidome. Twenty-four genomes were sequenced from strains selected to represent as much as possible the genetic diversity in C. novyi sensu lato. Sixty-one plasmids were identified in these genomes and 28 of them were completed. The genomic comparisons revealed four separate lineages, which did not strictly correlate with the species designations. The plasmids were categorized into 13 different plasmid groups on the basis of their similarity and conservation of plasmid replication or partitioning genes. The plasmid groups, lineages and species were to a large extent entwined because plasmids and toxin genes had moved across the lineage boundaries. This dynamic process appears to be primarily driven by phages. We here present a comprehensive characterization of the complex species group C. novyi sensu lato, explaining the intermixed genetic properties. This study also provides examples how the reorganization of the botulinum toxin and the novyi alpha toxin genes within the plasmidome has affected the pathogenesis of the strains.

Bacillary hemoglobinuria in dairy cows: clinical, hematological, biochemical, and pathological alterations

The present study was designed to describe the clinical, hematological, and biochemical changes as well as the pathological findings in dairy cows with bacillary hemoglobinuria. This study was conducted on 11 multiparous Friesian cows. Clinically, diseased cows exhibited severe fever, hemoglobinuria, and anorexia with variable degrees of respiratory distress. Hematological analyses revealed severe anemia, and biochemical analyses indicated disruption of hepatic function as serum activities of γ-glutamyl transferase and aspartate aminotransferase, and concentration of total bilirubin were significantly increased in diseased cows (P 0.05). Histopathological examination of liver specimens revealed multiple necrotic foci containing a large number of bacterial bacilli with accumulation of bilirubin within hepatic canaliculi. The lungs of dead cows were congested and hemorrhagic, and most blood vessels were impacted with bacilli. In the intestine and kidneys of dead cows, severe enteritis and severe nephrosis were observed, respectively. It can be concluded that bacillary hemoglobinuria is a fatal disease associated with many changes including clinical, hematological, biochemical, and pathological alterations that could be helpful for a rapid preliminary diagnosis of such disease.

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.

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.

An outbreak of bacillary haemoglobinuria in sheep in India.

An outbreak of bacillary haemoglobinuria was recorded in 60 out of 110 sheep in Ludhiana, Punjab, India. The condition was clinically characterised by fever, haemoglobinuria, constipation, weakness of hind quarters followed by recumbency, respiratory distress and death in 16 sheep. Haematological studies revealed moderate to severe degrees of anaemia associated with leucocytosis. Plasma gamma-glutamyl transferase, alkaline phosphatase and creatinine phosphokinase activities were significantly higher in haemoglobinuric sheep. Babesiosis and copper poisoning were ruled out on stained blood film examination and from blood mineral profiles, respectively. Post-mortem examination of affected sheep revealed no gross changes. Pure cultures of Clostridium haemolyticum isolated from heart blood, liver, kidney and spleen of freshly killed sheep confirmed the disease. Parenteral administration of procaine penicillin was effective in the treatment of affected sheep.

Bacillary haemoglobinuria diagnosis by the peroxidase-antiperoxidase (PAP) technique.

A peroxidase-antiperoxidase (PAP) technique was used to diagnose bacillary haemoglobinuria in formalin-fixed, paraffin-embedded liver tissues of cattle. The PAP method revealed Clostridium haemolyticum in the zone of liver necrosis characteristic of the disease and also in culture smears of this microorganism, but C. novyi type B, C. chauvoei, C. septicum and C. perfringens types B and C remained unstained by the PAP reaction. The PAP technique performed provides a specific, simple and rapid method to diagnose bacillary haemoglobinuria.

Bovine Bacillary Hemoglobinuria (Clostridium Haemolyticum) in Iran

Bacillary hemoglobinuria (redwater disease) in cattle is caused by Clostridium haemolyticum. The disease was first described in 1916 as an enzootic disease in areas of the western United States. Since then, the disease has been reported from Mexico, Peru, England, Chile, Turkey, New Zealand, and Australia. The toxicity of the organism for cattle, sheep, rabbits, mice, and guinea pigs has been reported. Investigations have been done on the biochemical, toxigenic, and immunologic aspects of the organism. For a decade starting in 1978, we investigated cases of icterohemoglobinuria in adult bovids at the Razi Institute, Tehran, Iran. The great majority of cases were leptospirosis and a few were either babesiosis or postparturient hemoglobinuria. Although Clostridium oedematiens type D had been isolated from necrotic liver lesions of a sheep, to the best of our knowledge there is no documented report on either bacillary hemoglobinuria or the isolation of Clostridium novyi type D from cattle in Iran. This is the first report on the occurrence of bacillary hemoglobinuria in Iran. In September 1988, a putrefied carcass of a dairy cow was submitted to the Pathology Department at the Razi Institute. The owner had 30 Holstein-native cross-bred cows on his farm, located 60 km west of Tehran near the Razi Institute. According to the owner, the cow had been ill for the previous 48 hours with anorexia and hemoglobinuria. A ne-