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Laboratory diagnosis of an Aspergillus fumigatus outbreak with concurrent secondary bacterial infection in a poultry farm in Jos, Plateau State, Nigeria

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Aspergillus fumigatus is recognized as a significant respiratory pathogen in poultry, causing substantial economic losses and severely impacting farmers’ livelihoods. This case report details the laboratory diagnosis of a clinical outbreak of A. fumigatus on a poultry farm in Jos, Plateau State, Nigeria. The diagnosis was made using a combination of historical data, clinical signs, postmortem examination, histopathology, and microbial culture, and identification. Notable findings included nodular lesions on the lungs and peritoneum, with fungal hyphae embedded within the lung tissue. Microbial culture revealed characteristic macroscopic features of A. fumigatus and the presence of Escherichia coli, indicating a potential secondary bacterial infection. Treatment involved administering 500,000 IU oral nystatin at 320 tablets per day for 14 days (1 tablet per 10 kg body weight) and Florum® (20% florfenicol) at 1mL per 2L of drinking water for 5 days. The farmer was advised to implement measures to enhance biosecurity, sanitation, and hygiene on the farm. Two weeks after treatment, the farmer recorded zero mortality even though he could not keep track of the actual number of dead birds, he however, estimated about 100 mortality. The farmer also acknowledged that the birds started laying at 17 weeks. A second farm visit when the birds were 19 weeks old also showed that they were thriving well and no mortality was recorded. This case report underscores the importance of accurate diagnosis for the early detection and prevention of disease agents and ensuring rational antimicrobial use for therapeutic purposes.
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Contact Asinamai Athliamai Bitrus bitrusaa@unijos.edu.ng Department of Veterinary Microbiology and Pathology, Faculty of
Veterinary Medicine, University Jos, Jos, Nigeria.


 Open Access
Laboratory diagnosis of an Aspergillus fumigatus outbreak with concurrent
secondary bacterial infecon in a poultry farm in Jos, Plateau State, Nigeria
Asinamai Athliamai Bitrus1,23,4, Oludotun Olubusola Oladele2,44,
1, Negedu Onogu Ameji4
1Department of Veterinary Microbiology and Pathology, Faculty of Veterinary Medicine, University of Jos, Jos, Nigeria
2
3
4
ABSTRACT
Aspergillus fumigatus         
        
case report details the laboratory diagnosis of a clinical outbreak of A. fumigatus on
             -
        
     

A. fumigatus and the presence of
Escherichia coli

®

       

          




ARTICLE HISTORY

Accepted June 02, 2024

KEYWORDS
Aspergillosis; diagnosis;
Escherichia coli; laboratory;
poultry; Plateau
Introducon

      -

     

     

-
 
     
     
fungus Aspergillus fumigatus   
A. fumigatus,A.
niger, A. nidulans, A. flavus
      
   Aspergillus fumigatus -
     




  -
-




23 jrvs

      

-

      
        

-
      
     
        -
    

 
     
       -
      -

Escherichia coli is
      
     
-

-
A. fumigatus 
       


9° 53’ 47.4972’’ N latitude and 8° 51’
29.9916’’ E longitude
      
      
-
  


-



      


Case Details and Diagnosc Plans
Clinical history



       
  

    ®   
-
  


®      




Necropsy
      

      
       
   
    
     

      
     

Histopathology
-
        
     
      
        -
     

      
      
      
     
     -
      
-
     
    
  
      

Microbial culture and suscepbility test

      
-
       
      
     
   
jrvs 24
A. A. Bitrus, A. R. Jambalang, O. O. Oladele, Y. J. Isa’ac, G. Y. Gurumyen, N. O. Ameji




      
     

      

-



as E. coli
      
 
        
     
-

A.
fumigatus-
       
      
         
          

     
-
     
E. coli and A. fumi-
gatus
   

     

       

     
      

A. fumigatus
-

-

    
  -
-

-
E. coliE. coli



Figure 1.            

Figure 2. -
   


25 jrvs

Treatment
    
  A. fumigatus   -
     E. coli 



®
     


     -
-

        
Figure 3.           
     

Figure 4. 
velvety colony morphology of A. fumigatus
Figure 5.   in vitro  -
      

jrvs 
A. A. Bitrus, A. R. Jambalang, O. O. Oladele, Y. J. Isa’ac, G. Y. Gurumyen, N. O. Ameji
        
        



Discussion

     
-
       

     
Aspergillus

     
-
Aspergillus 
Aspergillus
 A. fumigatus   -
Aspergillus fumigatus


  


Aspergillus     
    


       

    

     
-
 
     
      

   
        -

       

     
     
      
      
    

      
-
     -

-
        
     -
      
      
A. fumigatus
     


®
      


-
  -
      
   

       
       
    
 -

 
 
         
 -
     
      
       -
       
      -
     

Conclusion
     -
       
   E. coli   

   -
      
-
    -

Acknowledgment
     
     
27 jrvs

     
       
    
-


        

Conict of interest
-

Authors contribuon
        
     -
      

     
-


References
        
       


 

     
     
      

 
       

      
    
Aspergillus fumigatus
       

         
     Aspergillus fumigatus in
     

      

 

        
   
       
      

         
      

       
       

 
  
 

 
 
Escherichia coli    

        
      
    Escherichia
coli

       
       
  
     
   

       
   
     

   
    
Escherichia coli
     

 
        
    

      


        
      
    
  

 

Escherichia coli
      

         
 
     
       

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The present study was carried out to investigate the pathology of avian aspergillosis in commercial broiler chickens at Chittagong district. A total of 912 sick and dead chickens were collected from 20 commercial broiler farms and diagnosed for avian aspergillosis on the basis of clinical signs, symptoms and postmortem findings. The suspected birds were collected for necropsy examination and mycological culture. Gross lesions of multiple hard creamy to yellow colored, circumscribe plaques throughout the lungs surface and consolidated lung with necrotic areas were observed. Microscopically, the typical nodules consisted of caseous necrotic center were present. Identified the Aspergillus spp according to their color of colony growth on Potato Dextrose Agar media. The overall incidence of avian aspergillosis was found 6.14%. Among five Upazilla, significantly (p<0.007) higher and lower incidence was found in Patenga and Sitakunda that were 9.25% and 3.43% respectively. It was observed that highest incidence (8.22%) in rainy and lowest (3.16%) in winter but moderate (5.16%) in summer season. The disease was significantly (p<0.050) higher (8.27%) in age between 6-10 days and lower (4.11%) in age between 0-5 days. It was also found that incidence of avian aspergillosis was significantly (p<0.042) higher in flocks reared on sawdust litter (7.69%) as compared to rice husk litter (3. 46%).
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High and early mortalities of chicks often result in colossal economic losses. Aspergillosis, a fungal infection in chicks often occurs in the first 2 weeks of life and may be misdiagnosed as a common respiratory problem like Mycoplasmosis. Some Veterinarians are of the opinion that there is no effective way of treating Aspergillosis. This clinical case report describes the management of acute aspergillosis in chicks. Necropsies were done thrice. Grossly, congested livers, hemorrhagic nephritis, and congested lungs with whitish nodules were observed. Histologically, there were numerous multinuclear giant cells, macrophages and heterophils in the liver. The lung showed fungal hyphae and inflammatory cells. In the kidney, there was severe tubular epithelial necrosis indicated by the pyknotic nuclei, mononuclear infiltrates and haemorrhages. Considering the mortality rate, necropsy findings, microbial isolation and histopathologic observations, systemic Aspergillosis was diagnosed. The secondary bacterial infection was caused by Escherichia coli which was resistant to 8 antibiotics and susceptible only to Penstrep® (Penicillin +streptomycin). Initial resistance to copper sulphate necessitated a change to Nystatin which was used for 12 days via drinking water. Cumulative mortality was 757 (34.37%) in a flock of 2,202 chicks. Better control of systemic fungal infections may be enhanced by early application of anti-fungal susceptibility test.
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Confirmatory diagnosis of invasive aspergillosis is paramount to ensure proper treatment and effective management of the disease in food and companion animals. Suspected invasive aspergilllosis in chickens was encountered at post-mortem. Morphological and molecular methods were employed to identify Aspergillus from samples collected from dead chickens at post-mortem. Morphologically, two species of Aspergillus were identified, namely Aspergillus fumigatus and Aspergillus flavus. Molecular identification based on polymerase chain reaction (PCR) and sequence analysis of the partial 5.8 S rRNA, complete internal transcribed spacer-2 and partial 28S rRNA sequences bolstered morphological identification to arrive at the confirmatory diagnosis of the disease. Various hotspots that differentiate A. flavus from A. fumigatus and from other Aspergillus species were identified based on multiple sequence analysis. Maximum likelihood phylogenetic tree showed that isolates from the same species were grouped in the same clade. It is important to correctly identify the Aspergillus species in order to efficiently manage the disease.
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Avian colibacillosis is an acute and globally occurring infectious disease of domestic and wild birds caused by Escherichia coli, and it is associated with considerable economic losses mainly due to the morbidity and mortality associated. The present study aimed to describe the pathological, bacteriological and immunohistochemical aspects of avian colibacillosis in broiler chickens of Mozambique. Forty-nine broiler chicken presented anorexia, decreased weight gain, ataxia, diarrhea, dyspnea, and death in a clinical course of 3-5 days. The birds were raised in five farms (small, medium and large farms) with manual and automatic breeding system, with flocks ranging from 100 to 20,000 birds. At the necropsy, all birds had poor body condition, and the pericardium and the Glisson’s capsule of all avian exhibited different degrees of adherence often associated with severe fibrin deposition. The thoracic and abdominal air sacs were thickened and adhered to the costal wall. Mild, moderate or marked hepatomegaly associated with white pinpoint multifocal areas (100%, 49/49) and mild to moderate splenomegaly in 75.5% (37/49) with a mottled surface were observed. The lungs and kidney were enlarged and reddish. Histologically, a multiorgan fibrinoheterophilic polyserositis was observed in 75.5% of the cases (37/49), which were characterized by inflammatory infiltrates composed mainly of degenerative heterophils, macrophages and plasma cells, associated with fibrin deposits and intermixed by coccobacillary bacterial basophilic aggregates. These affected mainly the pericardium (28.6%, 14/49), the pleura (18.4%, 9/49), the Glisson’s capsule (10.2%, 5/49), the ventriculus (10.2, 5/33), and the proventriculus (8.2%, 4/49) serosa. Multifocal to coalescing areas of coagulative necrosis associated with similar inflammatory cells were observed mainly in the spleen (28.6%, 14/49), liver (24.5%, 12/49), and intestines (22.4%, 11/49). A similar infiltrate was also observed affecting the the lungs (16.3%, 8/49), the kidney (16.3%, 8/49) and the myocardium (14.3%, 7/49). Isolation and identification of E. coli was obtained in 12 cases through bacterial culture. Some organs (2 cases of each farms) were selected and submitted to immunohistochemistry anti-E. coli, and a positive stain was observed in all tested cases in liver (3/3), heart (4/4), spleen (1/1), lungs (4/4), intestines (4/4), bursa of Fabricius (1/1), ventriculus (1/1), and proventriculus (1/1) tissue sections. These results demonstrate that E. coli was the cause of mortality in these birds. Therefore, biosecurity and management measures should be employed to prevent and control the disease occurrence in Mozambique’s poultry farming.
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In this study, we aimed to determine the presence of pulmonary aspergillosis by histopathological and microbiological methods in geese that are economically grown in Kars region of Turkey. Totally 150 lung tissue samples of geese, an average age of 9 weeks, which died between 2013 and 2020 and were brought to our department were included in the study. Following the systemic necropsy paraffin blocks were prepared from the tissue samples and stained with Hematoxylin & Eosin (H&E) in order to detect histopathological changes. In order to reveal the presence of Aspergillus fungi, Periodic acid–Schiff (PAS) staining was applied to the sections as suggested by Facepath company. The microbiological examination of the tissue samples was carried out by the standard mycological culture technique on Sabouraud Dextrose Agar and by the phenotypical characterization of the emerged cultures. Various clinical symptoms such as difficulties in breathing, wheezing, weakness, anorexia, depression, diarrhea and nervous signs were recorded in geese. We observed large and small multifocal yellowish-white nodular structures in the lungs and air sacs macroscopically. In the histopathological examination of the lung tissues, we detected granulomatous structures with varying numbers and sizes. We diagnosed the Aspergillus agents in 20 (13.33%) of the tissue samples by detecting structures resembling typical tree branches in the middle of granulomatous structures with PAS staining. An identical positivity was obtained by the microbiological method and the emerged agent was solely identified as Aspergillus fumigatus with the growth pattern and macroscopic and microscopic morphological features. In conclusion, we found the presence of aspergillosis as 13.33% by histopathological and microbiological methods in geese which were brought to our department between 2013-2020. Based on these data, we concluded that aspergillosis is one of the most important infectious diseases among the goose deaths in Kars region of Turkey
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Aspergillosis is important in poultry production and human health but difficulty in its ante mortem diagnosis in addition to the unsuccessful outcome of most management options has made the disease a lingering nightmare to farmers. This study presents the clinicopathological features and management of aspergillosis in some poultry farms in Jos metropolis, Nigeria. Thirteen cases of aspergillosis from 12 poultry farms were confirmed and managed from April, 2019 to March, 2020 with 7.6% cases in a mixed turkey-broiler farm, 15.4% in broilers and 76.9% in layers. The ages affected ranged from 3 weeks to 21 weeks with 69.2% cases in pullet grower birds of 10 – 21 weeks compared to young chicks, 3 – 9 weeks with 30.8%. The rainy season accounted for 61.8% cases while mortality ranged from 1.0% to 17.4%. The clinical features were consistent and included dyspnoea; inappetence; retarded growth and mortality. While necropsy showed consolidated lungs with caseous nodules; liver with raised grayish or yellowish irregular nodules; peritonitis with the peritoneum and air sacs laced with caseous nodules; diphtheritic membrane and caseous nodules on intestinal mucosa; haemorrhagic bursitis with granulomata; caseous nodules on osseous tissues as well as nodules on the skin and skeletal muscles. Confirmatory diagnoses of aspergillosis were made by culturing nodules on Sabouraud’s Dextrose Agar. Antifungal susceptibility testing showed CuSO4 to be more susceptible which was used at the dosage of 3g/10L of drinking water for at least 7 days in all the cases. Concurrent bacterial infections were seen in some of the cases and were treated concurrently with antibiotics based on susceptibility testing while farmers using moldy feeds were asked to withdraw such feeds. Conclusively, aspergillosis is prevalent in Jos metropolis with severe impact on production and may be a silent killer disease in poultry due to constraints in diagnosis and its management. Keywords: Aspergillosis, clinicopathological features, Jos, management, Nigeria, poultry
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This paper describes a study carried out on people of different ages with various skin diseases who submitted themselves for examination and diagnoses at the Dermatophylosis Research Centre of the National Veterinary Research Institute, Vom, Plateau State, Nigeria. Samples of skin scrapping were collected and cultured on Sabouraud Dextrose Agar (SDA) while some were examined under the microscope for possible morphological identification of the fungi. The results showed that out of the thirty samples analyzed, nine genera of fungi were isolated comprising of Trichophyton 10 (33.3%), Aspergillus 9 (30%), Penicillum 4 (13.32%), Microsporum 2 (6.70%), Epidermorphyton 1 (3.33%), Trichosporum 1 (3.33%), Cryptococcus 1 (3.33%), Mucor 1 (3.33%), and Cephalosporium 1 (3.33). The study revealed high incidence of human fungal diseases, a situation that calls for good hygiene practices in the study area to mitigate and possibly eradicate the prevalence of human skin diseases in the rural area of Plateau State.
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Aim: The study was carried out to isolate and identify Aspergillus species from commercial birds with suspected aspergillosis in the poultry farms within Ado Ekiti metropolis Nigeria. Place and Period of Study: The study was carried out in the Department of Microbiology, Faculty of Science, Ekiti State University Ado Ekiti, Nigeria in August 2016. Methodology: A total of 35 sick/suspected birds were collected randomly from three poultry farms. At Ago-Aduloju poultry farms, 15 samples were randomly collected from 1000 birds while at Ekiti State University poultry farms, 10 samples were randomly collected from 500 birds. At Federal Polytechnic Ado Ekiti poultry farms, 10 samples were randomly collected from 700 birds. The bird’s selection was on the basis of clinical signs and symptoms such as difficulty in breathing, weight loss, drooping of wings and exercise intolerance. Swab samples were collected from each suspected/sick bird for mycological culture and molecular characterization of the isolates from each bird was carried out. The isolates were identified based on the color of the culture on Potato Dextrose Agar and microscopic examination. Molecular identification was done using 23S Ribosomal RNA Gene and Partial Sequence. Results: Six fungal strains that showed similar morphological and cultural characteristics of Aspergillus species were isolated. The isolates were coded ASP 1, ASP 2, ASP 3, ASP 4, ASP 5, and ASP 6. The identified organisms were; Aspergillus fumigatus qH 107 (ASP 1), Aspergillus fumigatus qH 107 (ASP 2), Aspergillus flavus M09 (ASP 3), Aspergillus flavus UOMS6 (ASP 4), Aspergillus fumigatus qH 107 (ASP 5), Aspergillus flavus qH 107 (ASP 6). Conclusion: It is evident that Aspergillus species were predominant in poultry farms selected in this study. Necessary precaution should be put in place to prevent the spread of aspergillosis. Poultry farmers are advised to avoid damp environments, moldy feeds, dry and dusty litters. Adequate ventilation should always be provided in poultry farms to prevent Aspergillosis.