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Histopathological Identification of Aspergillosis in Animals at Dhaka Zoo

Dr. Emdadul Haque Chowdhury
Professor
Department of Pathology, Bangladesh Agricultural University, Mymensingh-2202Bangladesh

S. A. Ahasan
Department of Pathology, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh

M. A. Rahman
Department of Livestock Services, Bangladesh; Krishi Khamar Sarak, Farmgate, Dhaka -1215, Bangladesh,

M. M. Rahman
Department of Livestock Services, Bangladesh; Krishi Khamar Sarak, Farmgate, Dhaka -1215, Bangladesh,

Abstract/ Executive Summary:

This study was conducted to investigate aspergillosis in animals at Dhaka Zoo to ascertain animal health, welfare and public health safety standard. One hundred and two necropsied tissue samples preserved in 10% neutral buffered formalin at necropsy from 36 animals of 25 different species were collected from Dhaka Zoo. Twenty five out of 36 study animals were suffering from granulomatous diseases. Among them 13 animals were suffering from Aspergillosis. Clinical history, nodular lesions from necropsy findings, granulomatous reactions along with fungal spores and characteristic radiating club on histopathology; dichotomously branching septate hyphae and mycelial conidiophore on special staining were revealed Aspergillosis in 13 animals of nine species that includes four rhesus macaques (Macaca mulatta), two samber deer (Cervus unicolor) and one of each species were nilgai (Boselaphus tragocamelus), horse (Equus caballus), stripped hyena (Hyena hyena), gayal (Bos frontalis), beisa oryx (Oryx beisa beisa), water buck (Kobus L. leche) and greater kudu (Tragelaphus strepsiceros). Present study provides evidence of existing Aspergillosis and similar long standing zoonotic diseases in majority of rest of the animals with health risk that shades health safety standard at Dhaka Zoo.

Keywords: Granuloma, Aspergillosis, Hyphae, Zoo animals, Histopathology

Location: Dhaka and Mymensingh

Start Date: 00-00-2011

End Date: 00-00-2012

Program Area: Animal Health and Management

Commodity: Animal

Objectives:

To investigate aspergillosis in animals at Dhaka Zoo to ascertain animal health, welfare and public health safety standard

Methodology:

The research work was conducted in the Department of Pathology, Bangladesh Agricultural University (BAU), Mymensingh-2202 and Dhaka Zoo, Bangladesh in 2011-2012. A total of 102 necropsied formalin-fixed tissue specimens from 36 animals of 25 different species at necropsy were investigated clinico-histopathologically and special staining technique was followed. The study comprised of eight rhesus macaques (Macaca mulatta), four spotted deers (Cervus axis/Axis axis), two samber deers (Cervus unicolor), two golden pheasant (Chrysolophus pictus), one of each species were guineapig (Cavia porcellus), wildebeest (Connochaetes taurinus), striped hyena (Hyena hyena), Indian/Asiatic lion (Panthera leo persica), gayal (Bos frontalis), American rhea (Rhea americana), Australian terrier dog (Canis lupus familiarizes), zebra (Equus zebra hartmannae), nilgai (Boselaphus tragocamelus), horse (Equus caballus), barking deer (Muntiacus muntjak), ostrich (Struthio camelus), crested serpent-eagle (Spilorins cheela), common languor (Presbytis entellus), fishing cat (Felis viverrina), beisa oryx (Oryx beisa beisa), reticulated python (Python molurus), water buck (Kobus L. leche), greater kudu (Tragelaphus strepsiceros), and olive baboon (Papio anubis). The animal belongs to non-human primates (n=10), carnivores (n=4), herbivores (n=16), reptiles (n=1) and birds (n=5). At necropsy, tissue changes were recorded; photographed and lesioned tissues of all vital organs with special attention to nodular lesions were preserved in 10% neutral buffered formalin. Formalin fixed samples were processed for paraffin embedding, sectioning and staining as routine hematoxylin and eosin staining (Luna, 1968), periodic acid Schiff (PAS) for staining fungus according to standard method of histopathological study (Mallory, 1968). In PAS staining, 0.5% Periodic acid solution and 0.2% Light Green solution (Stock) were used. Briefly tissue sections were deparaffinized and hydrated to distilled water before oxidized in periodic solution for 5 minutes followed by rinsing in distilled water then dipped in Schiff reagent solution for 15 minutes. It was then washed in running tap water for 10 minutes until a pink colour to develop followed by counter staining in light green for few seconds. Again washed in running tap water for 10 minutes then dehydrated in 95% alcohol, absolute alcohol, and cleared in xylene, two changes each followed by mounting with permount (McManus, 1948). Photomicrograph was taken using photomicrographic camera (Olympus PM-C 35 Model) and Digital Camera Mounted Photomicrographic device (Diffrential Interference Contrast – DIC) (Olympus, Nizol FC, E-5000, 8.4V, 0.9A, CE N 150), respectively.

Source of Information: J. Bangladesh Agril. Univ. 11(2): 265–270, 2013, ISSN 1810-3030

Article Link (Online Journal):

Funding Source:

1. Government

Budget:

Total Budget (Tk.) :

Achievement/Findings:

Pesent study revealed complex form of Aspergillosis concurrent with other granulomatous diseases in eight cases. It was noticed complexed with blastomycosis, coccidioidomycosis and cryptococcosis in each of single case along with concomitant with blastomycosis and coccidioidomycosis while four aspergillosis cases were noticed and complexed with coccidioidomycosis, cryptococcosis and histoplasmosis. Five cases of aspergillosis was identified complexed with mycobacteriosis. Without unnoticeable degree of variation, clinical history noted was almost similar to other works, nonetheless, it was difficult to differentiate among other granulomatous infections. Frequently observed clinical signs of coughing and anorexia were prominent; gradual emaciation followed by death while coughing and anorexia in cases of Aspergillosis was comparable to many other studies.Infrequently, non-correspondance to granulomatous reaction with nodular lesions was also observed. Most literature suggests the presence of foreign body giant cells in case of mycosis with the exception of findings by Jones et al. (1997) that suggests formation of both Langhan’s and foreign body giant cells in case of blastomycosis while present investigation showed presence of both type of giant cells in case of aspergillosis with huge number of Langhan’s type giant cells than foreign body. Aspergillosis in rhesus macaque, Samber deer and horse was reported, those are similar with the present study but data were not available incase of striped hyena, gayal, nilgai, beisa oryx, water buck and greater kudu to compare this study. PAS in this study showed characteristic dichotomously branching septate hyphae, occasional radiating clubs and mycelial conidiophore. Dhaka zoo animal might possess the infection from animal importation that were infected, dirty damp environment, contaminated or damp foods, and malnourishment or due to compromised immunity. This disease in Zoo animals can be confused with some other fungal diseases in particular blastomycosis, cryptococcosis, histoplasmosis, coccidioidomycosis, and mycobacteriosis by both their clinical manifestation and necropsy findings.However, further studies should be focused on typing and molecular characterization of aspergillus organism. Aspergillus organism possesses serious public health consequences and therefore, prevention and control of these diseases in Dhaka zoo should brought under notice to ensure health safety standard.

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