
Pathology and molecular characterization of recent Leucocytozoon caulleryi cases in layer flocks
Hae Rim Lee, Bon-Sang Koo, Eun-Ok Jeon, Moo-Sung Han, Kyung-Cheol Min, Seung Baek Lee, Yeonji Bae, In-Pil Mo
Journal of Biomedical Research ›› 2016, Vol. 30 ›› Issue (6) : 517-524.
Pathology and molecular characterization of recent Leucocytozoon caulleryi cases in layer flocks
Leucocytozoonosis was found in three layer farms in chickens with suspected fatty liver or fatty liver hemorrhagic syndrome in Korea between 2009 and 2011. These layer chicken flocks showed both mortality and decreased egg production for one or two weeks when they were between 59 and 82 weeks old. At the necropsy, the most prominent gross lesions were found in the liver, which was enlarged, had a fragile texture, exhibited yellowish discolorations, and had various hemorrhagic lesions. Tissue reactions associated with megaloschizonts specific for Leucocytozoon caulleryi were prominent upon microscopic examination of the liver without significant lipidosis. In addition, the ovaries and uterus were the most affected organs for Leucocytozoon caulleryi multiplication, which led to decreased egg productions. Molecular studies with formalin-fixed, paraffin-embedded tissues were performed in search of a partial region of the cytochrome b gene for hemosporidian parasites. Based on these results, the causal agent was determined to be closely related to Leucocytozoon caulleryi reported in Japan and Malaysia. In this study, we describe recently re-occurring leucocytozoonosis in layer chickens, which required histopathology for disease diagnosis. To prevent outbreaks and maintain chicken health and egg production, layer chickens need to be monitored for symptoms of leucocytozoonosis.
chicken leucocytozoonosis / Leucocytozoon caulleryi / layer chicken flocks / histopathology / megaloschizont
Fig.1 Gross lesions of chicken leucocytozoonosis cases suspected of fatty liver or fatty liver hemorrhagic syndrome in three chicken layer flocks between 2009 and 2011.A: The liver lobes from farm A were covered with blood clots (arrows). B: Liver from farm B was yellowish in color, and a subcapsular hemorrhagic region was present (arrow). C: Liver lobes from farm C were enlarged and had multifocal petechiae (arrows). D: The spleen from three flocks commonly showed enlargement and white mottling (arrows). E: The ovarian follicles were misshapen, atrophied, and congested (arrows). F: The kidney had multiple pin-point hemorrhages (arrows). |
Fig.2 Different stages of Leucocytozoon caulleryi megaloschizont degeneration observed in the heart.A: The intact forms of L. caulleryi megaloschizonts were observed as round, unilocular structures filled with numerous basophilic schizonts and surrounded by an eosinophilic capsular wall. These were observed in an aggregated form. B: Here, degenerating megaloschizonts had started to rupture, and merozoites were liberated from the defective schizonts (arrow). C: Partially or mostly depleted schizonts exhibited irregular capsules, and induced inflammatory cells and foreign-body giant cells (arrow) surrounded the defective structures. D: Capsular structures became more amorphous (arrow) and induced severe inflammatory reactions caused by lymphocytes, heterophils, foreign-body giant cells (open arrow), and macrophages infiltrating the periphery of the schizonts. HE staining. Bar= 50 mm (A), 100 mm (B), 50 mm (C), and 100 mm (D). |
Fig.3 Histopathological findings in the liver, spleen, lung, heart, kidney, pancreas, oviduct, and ovary from three cases of chicken leucocytozoonosis in different chicken layer farms between 2009 and 2011.A: Liver from farm A showed severe coagulative necrosis and eosinophilic amorphous residues of megaloschizont capsules of Leucocytozoon caulleryi surrounded by multi-nucleated giant cells (arrow). B: Liver from farm B showed a group of megaloschizonts specific for L. caulleryi, illustrating different degenerate stages (arrow) and pleomorphic lymphocytes infiltrated around hepatic veins (open arrow). C: Liver from flock C showed multifocal hemorrhages (arrows). D: The spleen showed lymphocytic depletion and eosinophilic residues of degenerating megaloschizonts (arrows). E: In the lung, megaloschizonts were found mostly in solitary forms in both the respiratory lobules and interlobular connective tissues (arrows), and defective schizonts induced mild pneumonitis (open arrow). F: In the kidney, there were hemorrhages (arrows) and interstitial nephritis associated with leucocytozoonosis. G: In the pancreas, the large areas of zymogen depletion in the exocrine cells were found around the groups of megaloschizonts (arrows) and were marginated from unaffected exocrine glands (open arrows). H: In the ovary, a considerable number of schizonts (arrow) multiplied and replaced the normal structure of the ovary. I: In the oviduct, numerous developing megaloschizonts were observed within the lamina propria of the gland, some of which invaded the muscularis layers. HE staining. Bar= 200 mm (A) and 500 mm (from (B) to (I)). |
Fig.4 A maximum-likelihood phylogenetic tree based on the partial Cytochrome b gene of various Leucocytozoon species in this study (asterisk) and those obtained from GenBank.All strains are indicated by the Leucocytozoon species name, the accession number, and the country where the agent was reported. Bootstrap values (>50%) are listed as percentages after 1,000 replications, and the genetic distances were calculated by the Kimura-two method. The scale bar indicates the number of substitutions per site. |
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