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Neonatal haemorrhagic diathesis in Belgium: gross pathology and cytology of blood and the haematopoietic system

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Abstract
Between September 2008 and September 2009, 25 calves with neonatal haemorrhagic diathesis (NHD) were autopsied. Breed distribution was 52% Belgian Blue and 48% Holstein Friesian. 47% was female and 53% male. The animals were on average 13 days old (7-21 days). Grossly, all cases showed generalized haemorrhages (petechiae, ecchymoses or haemorrhages) with variable lymphadenopathy. All calves had an anemic appearance. In 45% of the cases intestinal haemorrhage was present. Less frequent lesions were bacterial bronchopneumonia (23%), abomasal ulceration (32%), intestinal intussusception (4%) or icterus (4%). Histopathology of skin, oral mucosa, skeletal muscle, and hemorrhagic lesions in internal organs was performed and revealed multifocal extravasation of red blood cells (hemorrhage) with preservation of tissue architecture and absence of other abnormalities. Histopathological examination of the spleen and lymph nodes consistently showed severe lymphoid depletion in which T- and B-compartments were equally affected. Findings in the spleens were consistent with lymphocyte loss in the periarteriolar lymphoid sheets (PALS) and the lymphoid follicles. The red pulp was hypocellular with histiocytosis in a single case. Lymphnodes had a striking cortical and paracortical depletion of lymphocytes. In several cases, histiocytosis of the medullary sinuses was found. In 18% of the calves the thymus was examined and lymphoid depletion was present. The bone marrow was characterized by depletion of hematopoietic tissue in which all cell lines were affected with reduction of stem cells and progenitor cells including megakaryocytes (total aplasia). In 10 live calves bone marrow aspiration and cytology was performed at different stages of the disease. In three calves, the bone marrow punction was unsuccessful as only fat or subcortical tissue was present. A total aplastic bone marrow was seen in two calves in the terminal phase of the disease, with absence of megakaryocytes, lymphoid, myeloid and erythroid precursor cells. In the other 5 calves various amounts of cellular islands of precursor cells were still present and in two calves only a few megakaryocytes could be detected. In two animals the eosinophilic component was proportionally increased. In 5 animals groups of macrophages in close association with lymphocytes could be found in an aplastic bone marrow. The macrophages had an oval to round excentric nucleus with prominent nucleolus and fine stippled basophilic chromatin. The cytoplasm was abundant and vacuolated in two cases (consistent with activated macrophages). Hemophagocytosis was seen in two calves. In one calf, which eventually died of the disease, serial bone marrow aspiration was performed. On the onset of clinical symptoms (10 days old) the bone marrow was hypoplastic and myeloid precursor cells and megakaryocytes were still present. The eosinophilic component in the bone marrow was increased and a low number of macrophages was seen. In a second aspiration at an age of 14 days (the day before natural death) the bone marrow was totally aplastic with a sporadic presence of a myeloid precursor cell or macrophage. Cytology of peripheral blood showed stimulated lymphocytes, thrombocytopenia and granulocytopenia in most cases. In clinical cases no signs of regenerative anemia were seen. In a surviving calf normoblastosis and signs of regenerative anemia (Howel-Jolly bodies and reticulocytes) were seen 8 days after onset of clinical symptoms. Impression smears of the spleen were made in three calves, immediately after euthanasia. In all calves the lymphocytes were strongly activated (immunoblasts) and plasma cells were present. In one calf activated macrophages were seen. In another calf granulocytes were phagocytosing bacteria and in the third calf a sporadic eosinophil was seen. The findings are suggestive for an immune-mediated mechanism, of which the primary cause could be both toxic (natural toxins, medication, vaccination) or infectious.

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Chicago
Pardon, Bart, Veronique Saey, Jan Dierick, Griet Vercauteren, Kris De Clercq, Richard Ducatelle, Lennert Steukers, and Piet Deprez. 2009. “Neonatal Haemorrhagic Diathesis in Belgium: Gross Pathology and Cytology of Blood and the Haematopoietic System.” In Haemorrhagic Diathesis in Calves, Abstracts, ed. Claire Espinasse, 23–24. French Buiatrics Association.
APA
Pardon, B., Saey, V., Dierick, J., Vercauteren, G., De Clercq, K., Ducatelle, R., Steukers, L., et al. (2009). Neonatal haemorrhagic diathesis in Belgium: gross pathology and cytology of blood and the haematopoietic system. In C. Espinasse (Ed.), Haemorrhagic Diathesis in Calves, Abstracts (pp. 23–24). Presented at the Haemorrhagic Diathesis in Calves : satellite symposium of the 1st European Buiatrics Conference, French Buiatrics Association.
Vancouver
1.
Pardon B, Saey V, Dierick J, Vercauteren G, De Clercq K, Ducatelle R, et al. Neonatal haemorrhagic diathesis in Belgium: gross pathology and cytology of blood and the haematopoietic system. In: Espinasse C, editor. Haemorrhagic Diathesis in Calves, Abstracts. French Buiatrics Association; 2009. p. 23–4.
MLA
Pardon, Bart, Veronique Saey, Jan Dierick, et al. “Neonatal Haemorrhagic Diathesis in Belgium: Gross Pathology and Cytology of Blood and the Haematopoietic System.” Haemorrhagic Diathesis in Calves, Abstracts. Ed. Claire Espinasse. French Buiatrics Association, 2009. 23–24. Print.
@inproceedings{802069,
  abstract     = {Between September 2008 and September 2009, 25 calves with neonatal haemorrhagic diathesis (NHD) were autopsied. Breed distribution was 52% Belgian Blue and 48% Holstein Friesian. 47% was female and 53% male. The animals were on average 13 days old (7-21 days). Grossly, all cases showed generalized haemorrhages (petechiae, ecchymoses or haemorrhages) with variable lymphadenopathy. All calves had an anemic appearance. In 45% of the cases intestinal haemorrhage was present. Less frequent lesions were bacterial bronchopneumonia (23%), abomasal ulceration (32%), intestinal intussusception (4%) or icterus (4%). Histopathology of skin, oral mucosa, skeletal muscle, and hemorrhagic lesions in internal organs was performed and revealed multifocal extravasation of red blood cells (hemorrhage) with preservation of tissue architecture and absence of other abnormalities. Histopathological examination of the spleen and lymph nodes consistently showed severe lymphoid depletion in which T- and B-compartments were equally affected. Findings in the spleens were consistent with lymphocyte loss in the periarteriolar lymphoid sheets (PALS) and the lymphoid follicles. The red pulp was hypocellular with histiocytosis in a single case. Lymphnodes had a striking cortical and paracortical depletion of lymphocytes. In several cases, histiocytosis of the medullary sinuses was found. In 18% of the calves the thymus was examined and lymphoid depletion was present. The bone marrow was characterized by depletion of hematopoietic tissue in which all cell lines were affected with reduction of stem cells and progenitor cells including megakaryocytes (total aplasia). 
In 10 live calves bone marrow aspiration and cytology was performed at different stages of the disease. In three calves, the bone marrow punction was unsuccessful as only fat or subcortical tissue was present. A total aplastic bone marrow was seen in two calves in the terminal phase of the disease, with absence of megakaryocytes, lymphoid, myeloid and erythroid precursor cells. In the other 5 calves various amounts of cellular islands of precursor cells were still present and in two calves only a few megakaryocytes could be detected. In two animals the eosinophilic component was proportionally increased. In 5 animals groups of macrophages in close association with lymphocytes could be found in an aplastic bone marrow. The macrophages had an oval to round excentric nucleus with prominent nucleolus and fine stippled basophilic chromatin. The cytoplasm was abundant and vacuolated in two cases (consistent with activated macrophages). Hemophagocytosis was seen in two calves. In one calf, which eventually died of the disease, serial bone marrow aspiration was performed. On the onset of clinical symptoms (10 days old) the bone marrow was hypoplastic and myeloid precursor cells and megakaryocytes were still present. The eosinophilic component in the bone marrow was increased and a low number of macrophages was seen. In a second aspiration at an age of 14 days (the day before natural death) the bone marrow was totally aplastic with a sporadic presence of a myeloid precursor cell or macrophage. Cytology of peripheral blood showed stimulated lymphocytes, thrombocytopenia and granulocytopenia in most cases. In clinical cases no signs of regenerative anemia were seen. In a surviving calf normoblastosis and signs of regenerative anemia (Howel-Jolly bodies and reticulocytes) were seen 8 days after onset of clinical symptoms. Impression smears of the spleen were made in three calves, immediately after euthanasia. In all calves the lymphocytes were strongly activated (immunoblasts) and plasma cells were present. In one calf activated macrophages were seen. In another calf granulocytes were phagocytosing bacteria and in the third calf a sporadic eosinophil was seen. The findings are suggestive for an immune-mediated mechanism, of which the primary cause could be both toxic (natural toxins, medication, vaccination) or infectious.},
  author       = {Pardon, Bart and Saey, Veronique and Dierick, Jan  and Vercauteren, Griet and De Clercq, Kris and Ducatelle, Richard and Steukers, Lennert and Deprez, Piet},
  booktitle    = {Haemorrhagic Diathesis in Calves, Abstracts},
  editor       = {Espinasse, Claire},
  language     = {eng},
  location     = {Marseille, France},
  pages        = {23--24},
  publisher    = {French Buiatrics Association},
  title        = {Neonatal haemorrhagic diathesis in Belgium: gross pathology and cytology of blood and the haematopoietic system},
  year         = {2009},
}