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Differential Cytokine Production in Neonatal Ovine Lung in Response to Experimental Mannheimia haemolytica Infection
106
ESVP and ECVP Proceedings 2016
J. Comp. Path. 2017, Vol. 156, 54e141
BOID INCLUSION BODY DISEASE AND MYCOBACTERIOSIS CO-MORBIDITY IN A BOA CONSTRICTOR C. Jelinek *, U. Friedel y, E. Dervas *, A. Kipar * and U. Hetzel* *Institute of Veterinary Pathology and yInstitute of Bacteriology, Vetsuisse Faculty, University of Zurich, Switzerland Introduction: Boid inclusion body disease (BIBD) is a debilitating disease of captive boid snakes, associated with reptarenavirus infection. Affected animals often die from secondary bacterial infections, which are considered a consequence of immunosuppression due to the functional impairment of leucocytes that also contain the inclusion bodies (IBs). Here we present the case of a Boa constrictor with BIBD that also suffered from mycobacteriosis. Materials and Methods: An adult female Boa constrictor was humanely destroyed after the diagnosis of BIBD made on blood smears. A full post-mortem examination with histology and immunohistology for reptarenavirus was performed. A frozen liver sample was used for mycobacterial culture and identification by 16S sequencing. Results: The post-mortem examination revealed no gross findings apart from a mummified fetus. Histology identified the BIBD intracytoplasmic IBs in almost all cell types, and a multifocal granulomatous splenitis and hepatitis. Ziehl-Neelsen staining revealed low numbers of acid-fast bacteria within macrophages in the granulomatous infiltrates; these also exhibited IBs and were found to be reptarenavirus infected. The causative bacteria were identified as a member of the Mycobacterium simiae complex. Conclusions: M. simiae complex are environmental, non-tuberculous mycobacteria. In man, they generally induce lesions in patients with (infectious) co-morbidities, a scenario that might also apply to the present case, providing further evidence that BIBD is indeed immunosuppressive. Apparently, however, BIBD does not substantially affect basic leucocyte functions such as phagocytosis. No direct cause was found for the fetal death, but general malaise in the course of mycobacteriosis must be considered.
CYTOKINE RESPONSES IN OVINE LUNG FOLLOWING EXPOSURE TO BOVINE RESPIRATORY SYNCYTIAL VIRUS A.J. Masot *, A. G azquez *, A. Franco y and E. Redondo* *Laboratory of Histology and Pathology and yLaboratory of Anatomy, Animal Medicine Department, Caceres Veterinary Faculty, University of Extremadura, Spain Introduction: This study sought to determine the immunohistochemical expression of interleukin-1 beta (IL-1b), tumour necrosis factor alpha (TNF-a), interferon gamma (INF-g), IL- 4, IL-6, IL8, IL-10 and IL-12 and to measure the levels of these cytokines in lung tissue from lambs infected experimentally with BRSV. Materials and Methods: Lambs (n 5 15) were inoculated at 2 days of age with 20 ml of viral inoculum (1.26 106 TCID50/ml) or sterile media (n 5 15). Vital signs (i.e. rectal temperature, pulse and respiratory rates) were monitored daily in control and infected lambs. Lambs were killed and subjected to necropsy examination at 1, 3, 5, 7 and 15 days post inoculation. Results: Findings demonstrated a temporal association between pulmonary expression of these cytokines and lung pathology in BRSVinfected lambs. IL-4 and IL-10 were not primarily involved in the pathogenesis of BRSV infection in neonatal lambs. A significant increase in IL-1b, TNF-a, INF-g and IL-6 proteins and labelled cells was found, suggesting that these cytokines may play a major role in enhancing the biological response to BRSV, contributing to the development of lung lesions in BRSV-infected lambs. A significant increase of concentrations and number of immunolabelled cells for IL-8 and IL-12 was observed in infected lamb lungs throughout the study. Conclusions: Given the marked induction of IL-8 and IL-12, anticytokine agents targeting these inflammatory cytokines may be useful in the prevention and treatment of BRSV, in conjunction with measures to combat the causative pathogen and prophylactic methods aimed at preventing infection.
DIFFERENTIAL CYTOKINE PRODUCTION IN NEONATAL OVINE LUNG IN RESPONSE TO EXPERIMENTAL MANNHEIMIA HAEMOLYTICA INFECTION E. Redondo *, A. G azquez *, A. Franco y and A.J. Masot* *Department of Histology and Pathology and yAnimal Medicine Department, Veterinary Faculty, University of Extremadura, Spain Introduction: The immunohistochemical expression and the lung extract concentrations of interleukin (IL)-1b, tumour necrosis factor (TNF)-a and IL-8 in the lung of lambs infected experimentally with Mannheimia haemolytica (Mh) were investigated. Materials and Methods: The lambs were assigned randomly to two groups: infected and uninfected controls. The inoculum in each lamb of the infected group was 1.5 109 colony-forming units of Mh in 5 ml sterile nutrient broth. The control lambs were inoculated with 5 ml of sterile nutrient broth. The control and infected animals were killed from 1 to 15 days post infection (dpi). Results: The findings demonstrated a temporal association between pulmonary expression of these cytokines and lung pathology in ovine pulmonary pasteurellosis. Expression of IL-8 was much greater than that of TNF-a and IL-1b. Conclusions: The results of this study suggest that IL-1b, TNF-a and IL-8 inflammatory cytokines may play an important role in enhancing the biological response to Mh and contribute to the development of the lung lesions in ovine pulmonary pasteurellosis. IL-8 was the dominant inflammatory cytokine expressed within the lungs in ovine pasteurellosis; accordingly, anti-cytokine agents targeting this mediator may be most useful in the prevention and treatment of this disease, provided as a complementary measure to combat the causative pathogen agent, together with prophylactic measures to prevent the infections.
PATHOGENESIS OF PASTEURELLA MULTOCIDA IN RABBITS: LIGHT AND ELECTRON MICROSCOPICAL STUDIES S.H. Afifi *, S.K. Abd-Elghaffar * and G.M. Badry *Department of Pathology, Faculty of Veterinary Medicine, Assiut University, Assiut and yAnimal research Institute, Dokki, Cairo, Egypt Introduction: The variability in clinical signs of pasterurellosis in rabbits may be influenced by different Pasteurella multocida virulence factors. The objective of the present study was to describe the pathological changes associated with a virulent strain of P. multocida in rabbits by light and scanning electron microscopy. Materials and Methods: Sixteen rabbits (8e9 weeks old) were injected with 1 ml of 106 cfu/ml of bacteria in PBS. Two rabbits were inoculated with l ml PBS and used as a control. Samples from the nasal mucosa (both sides), middle part of trachea and from all the lobes of the lungs from both exposed and control groups were taken at 1, 2, 3, 4, 5, 6, 7 and 14 days post inoculation (dpi) and processed for light microscopy. The nasal mucosa at 14 dpi was processed for scanning electron microscopy (SEM). Results: The nasal mucosal changes were characterized by erosion of the nasal mucosa and hypertrophy of mucus glands at the 4th dpi and the submucosa had focal aggregation of lymphocytes. The tracheal changes included hyperplasia and hypertrophy of mucus cells and disorganization of cilia. The lungs showed features of fibrinous pneumonia and the presence of inflammatory cells during infection. Desquamation and attachment of bacteria to the nasal epithelium were observed by SEM. Conclusions: The capsule of P. multocida identified by SEM could be associated with the virulence of the bacterium. Acute fibrinous pneumonia and inflammatory cells were evident. Desquamation in the nasal cavity and attachment of bacteria were observed even at 14 dpi by SEM, suggesting rhinitis.
ESVP and ECVP Proceedings 2016
J. Comp. Path. 2017, Vol. 156, 54e141
BOID INCLUSION BODY DISEASE AND MYCOBACTERIOSIS CO-MORBIDITY IN A BOA CONSTRICTOR C. Jelinek *, U. Friedel y, E. Dervas *, A. Kipar * and U. Hetzel* *Institute of Veterinary Pathology and yInstitute of Bacteriology, Vetsuisse Faculty, University of Zurich, Switzerland Introduction: Boid inclusion body disease (BIBD) is a debilitating disease of captive boid snakes, associated with reptarenavirus infection. Affected animals often die from secondary bacterial infections, which are considered a consequence of immunosuppression due to the functional impairment of leucocytes that also contain the inclusion bodies (IBs). Here we present the case of a Boa constrictor with BIBD that also suffered from mycobacteriosis. Materials and Methods: An adult female Boa constrictor was humanely destroyed after the diagnosis of BIBD made on blood smears. A full post-mortem examination with histology and immunohistology for reptarenavirus was performed. A frozen liver sample was used for mycobacterial culture and identification by 16S sequencing. Results: The post-mortem examination revealed no gross findings apart from a mummified fetus. Histology identified the BIBD intracytoplasmic IBs in almost all cell types, and a multifocal granulomatous splenitis and hepatitis. Ziehl-Neelsen staining revealed low numbers of acid-fast bacteria within macrophages in the granulomatous infiltrates; these also exhibited IBs and were found to be reptarenavirus infected. The causative bacteria were identified as a member of the Mycobacterium simiae complex. Conclusions: M. simiae complex are environmental, non-tuberculous mycobacteria. In man, they generally induce lesions in patients with (infectious) co-morbidities, a scenario that might also apply to the present case, providing further evidence that BIBD is indeed immunosuppressive. Apparently, however, BIBD does not substantially affect basic leucocyte functions such as phagocytosis. No direct cause was found for the fetal death, but general malaise in the course of mycobacteriosis must be considered.
CYTOKINE RESPONSES IN OVINE LUNG FOLLOWING EXPOSURE TO BOVINE RESPIRATORY SYNCYTIAL VIRUS A.J. Masot *, A. G azquez *, A. Franco y and E. Redondo* *Laboratory of Histology and Pathology and yLaboratory of Anatomy, Animal Medicine Department, Caceres Veterinary Faculty, University of Extremadura, Spain Introduction: This study sought to determine the immunohistochemical expression of interleukin-1 beta (IL-1b), tumour necrosis factor alpha (TNF-a), interferon gamma (INF-g), IL- 4, IL-6, IL8, IL-10 and IL-12 and to measure the levels of these cytokines in lung tissue from lambs infected experimentally with BRSV. Materials and Methods: Lambs (n 5 15) were inoculated at 2 days of age with 20 ml of viral inoculum (1.26 106 TCID50/ml) or sterile media (n 5 15). Vital signs (i.e. rectal temperature, pulse and respiratory rates) were monitored daily in control and infected lambs. Lambs were killed and subjected to necropsy examination at 1, 3, 5, 7 and 15 days post inoculation. Results: Findings demonstrated a temporal association between pulmonary expression of these cytokines and lung pathology in BRSVinfected lambs. IL-4 and IL-10 were not primarily involved in the pathogenesis of BRSV infection in neonatal lambs. A significant increase in IL-1b, TNF-a, INF-g and IL-6 proteins and labelled cells was found, suggesting that these cytokines may play a major role in enhancing the biological response to BRSV, contributing to the development of lung lesions in BRSV-infected lambs. A significant increase of concentrations and number of immunolabelled cells for IL-8 and IL-12 was observed in infected lamb lungs throughout the study. Conclusions: Given the marked induction of IL-8 and IL-12, anticytokine agents targeting these inflammatory cytokines may be useful in the prevention and treatment of BRSV, in conjunction with measures to combat the causative pathogen and prophylactic methods aimed at preventing infection.
DIFFERENTIAL CYTOKINE PRODUCTION IN NEONATAL OVINE LUNG IN RESPONSE TO EXPERIMENTAL MANNHEIMIA HAEMOLYTICA INFECTION E. Redondo *, A. G azquez *, A. Franco y and A.J. Masot* *Department of Histology and Pathology and yAnimal Medicine Department, Veterinary Faculty, University of Extremadura, Spain Introduction: The immunohistochemical expression and the lung extract concentrations of interleukin (IL)-1b, tumour necrosis factor (TNF)-a and IL-8 in the lung of lambs infected experimentally with Mannheimia haemolytica (Mh) were investigated. Materials and Methods: The lambs were assigned randomly to two groups: infected and uninfected controls. The inoculum in each lamb of the infected group was 1.5 109 colony-forming units of Mh in 5 ml sterile nutrient broth. The control lambs were inoculated with 5 ml of sterile nutrient broth. The control and infected animals were killed from 1 to 15 days post infection (dpi). Results: The findings demonstrated a temporal association between pulmonary expression of these cytokines and lung pathology in ovine pulmonary pasteurellosis. Expression of IL-8 was much greater than that of TNF-a and IL-1b. Conclusions: The results of this study suggest that IL-1b, TNF-a and IL-8 inflammatory cytokines may play an important role in enhancing the biological response to Mh and contribute to the development of the lung lesions in ovine pulmonary pasteurellosis. IL-8 was the dominant inflammatory cytokine expressed within the lungs in ovine pasteurellosis; accordingly, anti-cytokine agents targeting this mediator may be most useful in the prevention and treatment of this disease, provided as a complementary measure to combat the causative pathogen agent, together with prophylactic measures to prevent the infections.
PATHOGENESIS OF PASTEURELLA MULTOCIDA IN RABBITS: LIGHT AND ELECTRON MICROSCOPICAL STUDIES S.H. Afifi *, S.K. Abd-Elghaffar * and G.M. Badry *Department of Pathology, Faculty of Veterinary Medicine, Assiut University, Assiut and yAnimal research Institute, Dokki, Cairo, Egypt Introduction: The variability in clinical signs of pasterurellosis in rabbits may be influenced by different Pasteurella multocida virulence factors. The objective of the present study was to describe the pathological changes associated with a virulent strain of P. multocida in rabbits by light and scanning electron microscopy. Materials and Methods: Sixteen rabbits (8e9 weeks old) were injected with 1 ml of 106 cfu/ml of bacteria in PBS. Two rabbits were inoculated with l ml PBS and used as a control. Samples from the nasal mucosa (both sides), middle part of trachea and from all the lobes of the lungs from both exposed and control groups were taken at 1, 2, 3, 4, 5, 6, 7 and 14 days post inoculation (dpi) and processed for light microscopy. The nasal mucosa at 14 dpi was processed for scanning electron microscopy (SEM). Results: The nasal mucosal changes were characterized by erosion of the nasal mucosa and hypertrophy of mucus glands at the 4th dpi and the submucosa had focal aggregation of lymphocytes. The tracheal changes included hyperplasia and hypertrophy of mucus cells and disorganization of cilia. The lungs showed features of fibrinous pneumonia and the presence of inflammatory cells during infection. Desquamation and attachment of bacteria to the nasal epithelium were observed by SEM. Conclusions: The capsule of P. multocida identified by SEM could be associated with the virulence of the bacterium. Acute fibrinous pneumonia and inflammatory cells were evident. Desquamation in the nasal cavity and attachment of bacteria were observed even at 14 dpi by SEM, suggesting rhinitis.