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Adhesive tape preparation from air sac culture. A single typical conidiophore terminating in a flask-shaped vesicle bearing uniseriate phialides characteristic of Aspergillus fumigatus . Bar=20 μm. Inset: Tease mount preparation from air sac culture. Uniseriate phialides and smooth spores are appreciated. Bar=20 μm.
Source publication
We present the first case of angioinvasive pulmonary aspergillosis caused by Aspergillus fumigatus in a
Himalayan Griffon vulture (Gyps himalayensis). We describe in detail the clinical, gross, histopathological
and mycological findings. Clinical signs of weakness, emaciation, dyspnea, incoordination, and inability to fly
were observed in this bird...
Context in source publication
Context 1
... trachea, pericardium, aorta, pulmonary artery and kidneys. The nodules were of various sizes ranging between 1 and 3 mm in diameter, and the surface was smooth, shiny and convex. Nodules were also seen embedded in the parenchyma of the lungs, and occasional foci of necrosis and consolidation of lung tissue were also found. Abdominal air sacs were highly thickened wrinkled and the luminal surface was fuzzy and green in colour ( Figure 1). The tracheal lumen contained thick yellowish exudate in the anterior portion. Multifocal patchy areas of necrosis were found on the surface of the liver. Nodules were crushed, treated with 10% KOH and stained with lactophenol cotton blue stain; segments of branching fungal hyphae with dichotomous branching were observed under the microscope indicative of Aspergillus spp. The same could be observed from the scrapings of the luminal wall of affected air sacs. Nodules and tissue samples were taken for histopathology and fungal culture. A tentative diagnosis of Aspergillus fumigatus infection (aspergillosis) was made. Nodules, lung tissue triturate and air sac smears were inoculated into Sabouraud dextrose agar (SDA, HiMedia) with chloramphenicol (0.05 mg mL -1 ) and incubated at 37°C for seven days. The grown colonies were identified based on their macro- and micro-morphological features. Tease mount and adhesive tape wet preparations with or without lactophenol cotton blue were made for microscopic observation as described previously (Quinn et al ., 2004; McClenny, 2005; Johnson and Borman, 2010). The tissue samples were fixed in 10% neutral buffered formalin for histopathology. Briefly, tissues were washed overnight in tap water, and dehydrated in ascending grades of alcohol before passing through three changes of xylene. Dehydrated tissues were embedded in paraffin wax. Formalin-fixed paraffin-embedded tissues were cut to get sections with a thickness of 5 μm, and stained with haematoxylin and eosin. Microscopic examination of thickened air sacs revealed granulomatous airsacculitis with severe fibrosis and thickening of air sac wall. Multifocal non-encapsulated granulomas were observed in the lung parenchyma (superficial or deep) which had central necrotic cellular debris, surrounded by a zone of mixed inflammatory cell population including heterophils, lymphocytes and histiocytes, and radiating fungal hyphae having parallel walls at the periphery of the granuloma . Giant cells could not be observed in any of the lesions. Granulomas were also present on the surface of trachea, aorta, pulmonary artery and kidneys causing architectural disturbance. Radially arranged fungal hyphae without structured granuloma were also noticed (Figure 2) in lung parenchyma and the lumen of parabronchioles, as described by Nardoni et al . (2006) and Cacciuttolo et al . (2009). There was vascular invasion by fungal hyphae involving numerous small to large veins of lungs and air sacs (Figure 3). Frequently, fungal hyphae could be observed on the wall and/or in the lumen of many blood vessels (Figure 4). A mixed inflammatory cell reaction was noticed intramurally in affected blood vessels resulting in a thrombus obstructing the blood vessel lumen. Penetrating fungal hyphae caused severe damage to both alveolar epithelium and the blood vessel wall. Numerous thrombosed vessels were observed as a result of fungal hyphae invasion and intramural host reaction. The liver showed focal areas of hepatocyte degeneration and necrosis, and the Kupffer cells were prominent with abundant hemosiderin. Lymphoid depletion and reticular cell hyperplasia were characteristic in the spleen. Inoculation of samples in SDA resulted in exuberant growth of fungi at 37°C; a single colony covered the entire standard petri dish in five days. The fungal colony had bluish green or dark green centre with white periphery, and the reverse side of the colony was cream in colour. The texture was floccose initially, and turned granular after five days of incubation. Microscopically, tease mount or adhesive tape wet preparations revealed upright smooth conidiophores resulting in flask-shaped vesicles bearing uniseriate-phialides producing smooth round spores characteristic of Aspergillus fumigatus (Figure 5) (Quinn et al ., 2004; Johnson and Borman, ...
Citations
... Aspergillus spp. are known to have intense angiotropism so vascular lesions such as aneurysms and thromboembolism are often observed in these cases (Barathidasan et al., 2013;Veiga et al., 2021). Intravascular hyphae in the lung and rib with vasculitis demonstrate this tropism, and if this bird had survived, disseminated aspergillosis could have developed later in course of infection. ...
Aspergillosis is a fungal disease with high morbidity and mortality in wild and exotic bird species. The aim of this report is to describe a case of acute aspergillosis caused by Aspergillus fumigatus in a young African grey parrot (Psittacus erithacus). A three-month-old male African grey parrot from a commercial breeding site presented serous nasal discharge, inappetence, and wheezing on pulmonary auscultation. The parrot died three days after the onset of clinical signs. Postmortem evaluation demonstrated multiple smooth gray plaques in air sacs and left lung with dark red areas and adhesions to the ribs. Microscopically, there were intralesional hyphae and conidiophores in the lungs, air sacs, and ribs, which were associated with pneumonia, airsacculitis, and osteomyelitis, respectively. DNA samples were extracted from paraffinized tissues and subjected to PCR targeting the ITS-2 region, followed by sequencing, which yielded a sequence with 100% coverage and 100% identity to Aspergillus fumigatus sequences. Although A. fumigatus infection is quite common in birds, a particular aspect of interest in this case was the finding of conidiophores in the bone marrow, which may occur in birds due to air circulation through pneumatic bones.
Keywords:
Aspergillus sp.; mycoses; fungus; Psittacidae; paraffinized tissue; sequencing
... Breeding birds are at an increased risk of bacterial infections (Sonia González-Braojos et al., 2012). For example, Aspergillus fumigatus, isolated from nests, cause fungal infections in birds' lungs and air sacs, and birds are particularly susceptible to these infections (Barathidasan et al., 2013). The factors affecting the microbial colonization in the nest are not clear; nest structure, nesting materials, and the adult bird's growth may affect the richness of the microbial communities (Kornillowicz and Kitowski, 2018;Costanzo et al., 2022), and ectoparasites may also affect the bacterial environment of bird nests (Tomás et al., 2018). ...
... Costanzo et al. (2022) found that the microbiota in the nest may affect the gut microbes of the lesser kestrel (Falco naumanni). A variety of potentially pathogenic microbiota were also isolated in the nests of alpine vulture (Gyps himalayensis), warbler (Troglodytes aedon), and some wetland birds, which may affect chick development (David et al., 1998;Barathidasan et al., 2013;Korniłłowicz and Kitowski, 2013). In addition, the bacterial communities on the eggshell may also influence the hatching success and the status of the chicks (Peralta-Sánchez et al., 2012;Song et al., 2023). ...
... Additionally Pseudomonas was also found in the feathers of the eastern blue bird (Sialia sialis) and the feces of other wild birds (Brittingham et al., 1998;Shawkey et al., 2005). Furthermore, we detected a small amount of Aspergillus (1.79%) in used nests, a common avian pathogenic bacterium that caused fungal infection in the lungs and airbags (Barathidasan et al., 2013). Aspergillus had been found in the intestines and feces of birds such as Japanese quail (Coturnix japonica) (Huff et al., 1992), wild turkey (Meleagris gallopavo) (Quist et al., 2000), and house sparrow (Passer domesticus) (Lawson et al., 2006). ...
Nest microbiota plays a vital role in the breeding and development of birds, which not only provides protection to bird hosts but also negatively affects the host. At present, it is unclear whether the composition of the microbes in the nests is affected by nesting. For this reason, we hung artificial nest boxes to simulate the natural nesting environment and combined 16S rRNA and ITS high-throughput sequencing technology to further study the differences in microbial composition and richness between used nests and control nests of Japanese tits (Parus minor). The study found that the bacteria in used nests and control nests showed significant differences at the phylum level (p < 0.05). It is also worth noting that the predominant bacteria in used nests were Proteobacteria (51.37%), Actinobacteria (29.72%), Bacteroidetes (6.59%), and Firmicutes (3.82%), while the predominant bacteria in control nests were Proteobacteria (93.70%), Bacteroidetes (2.33%), and Acidobacteria (2.06%). Both used nests and control nests showed similar fungi at the phylum level, which consisted mainly of Ascomycota and Basidiomycota, although significant differences were found in their relative abundance between both groups. The results of alpha diversity analysis showed significant differences in bacteria between the two groups and not in fungi. However, the beta diversity analysis showed significant differences between both bacteria and fungi. In summary, our results showed that the used nests had a higher abundance of beneficial microbiota and a lower presence of pathogenic microbiota. Therefore, we speculate that birds will change the characteristics of the nest microbial composition in the process of nest breeding to ensure their smooth reproductive development.
... Fungi of the genus Aspergillus sp. and Candida sp. were found concomitantly associated with avian pox lesions, characterizing themselves as superficial cutaneous mycoses. The clinical presentation as a superficial cutaneous mycosis differs from what is usually observed in cases of infection by these agents due to its angioinvasive character, which contributes to systemic involvement in birds (Ceolin et al. 2012, Barathidasan et al. 2013. Secondary fungal infections caused by Candida sp. can induce serious lesions in Lovebirds (Agapornis roseicollis) and Golden Eagles (Aquila chrysaetos), leading to pulmonary granulomas and encephalitis (Tsai et al. 1997, Shrubsolecockwill et al. 2010. ...
This study describes the epidemiological and clinical-pathological aspects of outbreaks of avian pox diagnosed in poultry associated with fungal co-infections. A retrospective study was carried out, and cases of avian pox with suspected associated fungal infection were selected. The slides were subjected to routine and special histochemical staining in cases of suspected fungal infection, in addition to performing the immunohistochemical technique. Macroscopically, there were nodular lesions with crust formation, and histologically, the lesions were characterized by hyperplasia of the spinous layer associated with multiple eosinophilic intracytoplasmic inclusions. In seven cases, morphologically compatible structures with fungi were verified through black impregnation with GMS and strongly stained in pink with PAS. The morphotintorial aspects were suggestive of fungi belonging to the genus Aspergillus sp. and Candida sp. The diagnosis of fungal co-infections was confirmed through immunohistochemistry, with positive immunostaining for fungi of the genus Aspergillus sp. (five cases) and Candida sp. (two cases). It is concluded that the occurrence of fungal co-infections secondary to the cutaneous lesions of avian pox represents a complicating factor of the disease, favoring the weakness of these animals and death. Therefore, the investigation of associated secondary agents is necessary.
... FaĴ y changes, cloudy swelling, necrosis and infi ltration of red blood cells are seen in aspergillosis aff ecting liver 256 . In angio-invasive pulmonary, vascular invasion by fungal hyphae involving numerous small to large veins of lungs and air sacs and numerous thrombosed vessels are characteristic 257 . ...
... Ducks are reported to be the most susceptible poultry species to AF, followed by turkeys, broilers and laying hens, with ducks being approximately 200 times more 257 Punnoose et al. sensitive than chicks, especially for acute hepatotoxic eff ects 260,261 . Ducklings exhibit 100 percent mortality at 1 ppm AF 262 , and ducks are the only poultry species that develop hepatocellular carcinoma upon AFB1 exposure 260 . ...
... Zmiany patologiczne w przebiegu aspergilozy są zróżnicowane w zależności od postaci choroby (23,31,32,37,42). Ptaki z ostrą aspergilozą są na ogół w ogólnej dobrej kondycji zdrowotnej (43,44), podczas gdy wyniszczenie z zanikiem mięśni piersiowych i znikomą warstwą tłuszczu podskórnego oraz odwodnienie są cechami zakażenia przewlekłego (45,46,47). Horowitz i wsp. ...
... konidioforów, zarodników i strzępek, wysięk zapalny i liczne komórki odpornościowe w drogach oddechowych i wokół nich to wspólne cechy wszystkich postaci aspergilozy (38,44). Inwazja grzybicza w płucach powoduje krwotoki, zakrzepicę naczyń, zawał tkanek i przypuszczalne rozprzestrzenianie Aspergillus do odległych narządów (47,54). Ściany worków powietrznych są gęsto pokryte przez nacieki zawierające fibrynę, heterofile i makrofagi, natomiast ich powierzchnia jest skolonizowana przez grzybnię ze sporadycznie występującymi konidiami (50,55). ...
Aspergillosis holds a very special place in veterinary and human medicine,
because it is the main type of mycoses, affecting birds and mammals, including
humans. The objective of this review was to synthesize the current knowledge
of aspergillosis in wild avifauna in the context of the pathogenesis, diagnosis,
and treatment of this infection. Aspergillosis typically occurs after inhalation of
ubiquitously available spores, but localized infections of the eye or skin are also
possible. Acute disease may occur following exposure to a substantial number
of spores from a point source. The more chronic forms are slowly progressing
infections that affect birds showing some degree of immunodeficiency and may
result from regular inhalation of spores. Aspergillosis usually presents with the
development of progressive and severe dyspnea with gasping, accelerated openmouth breathing, tail bobbing, and sometimes a non-productive cough. Gurgle,
rales, or wheezy sounds and a change in the voice may be heard in cases of
mycotic tracheitis. Treatment of avian aspergillosis, when possible, is not always
successful because of the often advanced stage of the disease when the diagnosis
is confirmed, the lack of pharmacokinetic data on antifungal drugs in most avian
species, the failure of drugs to penetrate target tissues, and the frequent presence
of concurrent diseases and immunosuppression. An oral solution of itraconazole
has recently been registered as the first antifungal product for ornamental birds
in Europe. Moreover, aspergillosis prevention measures are based on two main
axes: controlling the level of exposure and minimizing the stressors
... In the recent past, we have recorded sporadic mortality due to acute as well as chronic form of aspergillosis in several captive and free ranging pigeons, peafowls and Griffon vultures particularly during winter periods. Despite the potential significance of Aspergillus infection and associated mortality, only one case report of IPA in Himalayan vulture has been reported from India (26). To the author's best knowledge, this communication is the first recorded evidence from India that describes anatomopathological features of acute and chronic invasive pulmonary aspergillosis (AIPA and CIPA) in Pavo cristatus (pea fowls), Columba livia (pigeons), and Gyps fulvus (Griffon vultures) with an attempt to grade the lesions based on gross and histopathology. ...
... In the recent past, we have recorded cases of aspergillosis domestic birds including poultry, Japanese quails, emu and turkeys (unpublished) and now in few wild birds such as vultures, pigeons and peafowls (26,52). Although the fungal species A. fumigatus was consistently isolated from all the birds, the anatomopathological features were distinct among different kinds of avian host. ...
Aspergillosis, caused by Aspergillus fumigatus, a fungal pathogen, can vary from localized infection to severe life threatening invasive or disseminated systemic diseases in birds. The objective of this disease investigation is to evaluate and grade the anatomopathological disparity in the cases of invasive pulmonary aspergillosis (IPA) in Columba livia (pigeons) (3), Pavo cristatus (pea fowls) (2) and Gyps fulvus (Griffon vultures) (2). Necropsy gross lesions varied from mere congestion of lungs in P. cristatus, congestion and large necrotizing masses surrounded by zone of hyperemia (10 mm dia) in lungs of C. livia and typical disseminated granuloma in the lungs, airsacs and organs of other serous membranes in G. fulvus. Histopathology varied from extensive parenchymal necrosis amidst exuberant fungal invasion in P. cristatus, multifocal to focally extensive tissue necrosis with colonies of fungal hyphae surrounded by heterophils and lymphocytes in C. livia, and typical mycotic granuloma embedded in the lungs, airsacs and thoraco-abdominal serous membranes with angio-invasion in G. fulvus. Based on gross and histopathological findings, we diagnosed the cases as Acute Invasive Pulmonary Aspergillosis (AIPA) in peafowls and pigeons, and Chronic Invasive Pulmonary Aspergillosis (CIPA) in Griffon vultures. Very few case reports are available on aspergillosis in wild avian species, and this report is to document cases of IPA in peafowls, pigeons and vultures. This is the first report of its kind which evaluates anatomopathological disparity of IPA in pigeons, peafowls and vultures with a proposed anatomopathological grading system which would help to understand and investigate the nature of aspergillosis in different avian hosts.
... Birds that succumb to acute aspergillosis are generally in good flesh condition [170,171], while wasting (pectoral muscle atrophy and negligible subcutaneous and internal fat) and dehydration are common features of chronic infection [97,98,134,172]. Emaciation has been correlated with the progression of the disease in common loons sheltered for rehabilitation [140]. ...
... The severity and the degree of development of the disease determine both the morphology and extension of macroscopic lesions along the coelomic serosa [102]. Gross lesions, either alone or in association (invasive aspergillosis) with others, have been observed in/on the brain, kidneys, liver, spleen, intestine, testis, bones, pericardium, and aorta [13,35,36,44,88,103,128,166,167,172,[175][176][177][178][179]. The initial phase is characterized by hemorrhagic and edematous lesions that progress to a granulomatous type of inflammation [140]. ...
... The accumulation of various fungal elements (conidiophores, spores, hyphae), inflammatory exudate, and cells in and around airways are common features [28,44,171]. Fungal angioinvasion results in hemorrhages, vascular thrombosis, tissue infarction, and putative dissemination of Aspergillus to distant organs [23,172,182]. Air sac walls are diffusely thickened by infiltrates containing fibrin, heterophils, and macrophages, whereas their surface is colonized by mycelium with occasional conidial heads [17,98,131]. ...
The ubiquitous fungi belonging to the genus Aspergillus are able to proliferate in a large number of environments on organic substrates. The spores of these opportunistic pathogens, when inhaled, can cause serious and often fatal infections in a wide variety of captive and free-roaming wild birds. The relative importance of innate immunity and the level of exposure in the development of the disease can vary considerably between avian species and epidemiological situations. Given the low efficacy of therapeutic treatments, it is essential that breeders or avian practitioners know the conditions that favor the emergence of Aspergillosis in order to put adequate preventive measures in place.
... While aspergillosis is a common problem in captive individuals (Chege et al. 2013), some studies in wild Cinereous and Himalayan Gyps himalayensis Vultures have reported cases of pneumonia due to Aspergillus spp. probably associated with concomitant pathologies that produce an immune alteration, suggesting that this fungus could be an opportunistic pathogen for vultures that should be evaluated (Jung et al. 2009, Barathidasan et al. 2013. Although there is little information about isolations of mycotic microorganisms in wild vultures, the few articles we found suggest that opportunistic mycosis could be a potential threat for vulture populations producing disease, in some cases accompanied by severe lesions such as oral ulcers or pneumonia (Jung et al. 2009, Barathidasan et al. 2013, Pitarch et al. 2017. ...
... probably associated with concomitant pathologies that produce an immune alteration, suggesting that this fungus could be an opportunistic pathogen for vultures that should be evaluated (Jung et al. 2009, Barathidasan et al. 2013. Although there is little information about isolations of mycotic microorganisms in wild vultures, the few articles we found suggest that opportunistic mycosis could be a potential threat for vulture populations producing disease, in some cases accompanied by severe lesions such as oral ulcers or pneumonia (Jung et al. 2009, Barathidasan et al. 2013, Pitarch et al. 2017. ...
The effects that microorganisms (bacteria, viruses and fungi) have on their hosts remain unexplored for most vulture species. This is especially relevant for vultures, as their diet consists of carcasses in various stages of decomposition, which are breeding grounds for potentially pathogenic microorganisms. Here we review current knowledge of bacterial, viral and mycotic microorganisms present in wild vultures. We consider their potential to cause disease in vultures and whether this poses any population‐level threats. Furthermore, we address the question of whether vultures may act as disease spreaders or mitigators. We found 76 articles concerning bacterial, viral and mycotic microorganisms present in 13 vulture species, 57 evaluating bacteria, 13 evaluating viruses and six evaluating mycotic microorganisms. These studies come from all continents where vultures are present, but mainly from Europe and North America, and the most studied species was the Griffon Vulture Gyps fulvus. We found that vultures are colonized by zoonotic pathogens, and even host‐specific human pathogens. Some recorded bacteria showed multi‐antibiotic resistance, especially those that can be associated with anthropogenic food subsides such as supplementary feeding stations. We found evidence that vulture health can be affected by some microorganisms, producing a wide array of clinical alterations that have the potential to influence mortality risk and fitness. We did not find clear scientific evidence that vultures play an epidemiological role spreading microorganisms to humans and other species. However, there is evidence that vultures could prevent the spread of infectious diseases through their removal of decomposing organic material. The evaluation of vulture exposure to microorganisms is of fundamental importance to design better conservation policies for this threatened group, which may serve a key role as ecosystem cleaners.
... These lesions had been mainly associated with chronic courses of avian aspergillosis (Redig, 1993(Redig, , 2007, but should be considered in acute and subacute courses as well. The involvement of these organs may hint at an invasive form of aspergillosis with spreading per continuitatem, per contiguitatem, and haematogenous spreading as reported in other bird species (Richard & Thurston, 1983;Cacciuttolo et al., 2009;Barathidasan et al., 2013). In general, the occurrence of pathological lesions and positive results in histology and mycology correlated significantly with the ID. ...
Aspergillosis is a common and life-threatening respiratory disease in raptors with acute and chronic courses. Among raptors, gyr falcons (Falco rusticolus) and their hybrids are often declared to be highly susceptible with juvenile individuals being the most susceptible. However, species and age specific experimental studies are lacking and minimal infective doses for Aspergillus spp. conidia are unknown.
Therefore, 8-week old, healthy gyr hybrid falcons (F. rusticolus X F. cherrug) (N=18) were experimentally infected with Aspergillus fumigatus using a single intratracheal inoculation with varying dosages of conidia (10² to 10⁷ conidia).
Over 28 days, clinical signs were monitored as well as hematological and serological parameters. Following euthanasia, necropsy, histopathology, bacteriology and mycology were performed. Re-isolated fungi were compared to the inoculum using microsatellite length polymorphisms.
During the trial, clinical signs and dyspnea correlated significantly with the infective dose. Necropsy revealed fungal lesions in the upper and lower airways of 10/18 inoculated falcons, but not in the control birds. In 9/18 inoculated falcons fungal granulomas were confirmed in histopathology and A. fumigatus was re-isolated from these granulomas. Except one nasal isolate all re-isolated fungal strains were identical to the inoculum strain. Based on mycology and histopathology a minimal infective dose 50 percent was calculated to be MID50% (±S.E.) = 104.52±0.67 for a single tracheal inoculation of A. fumigatus conidia.
This study demonstrates for the first time that a single exposure is able to cause acute aspergillosis in juvenile falcons.
... Within zoopathogenic fungi the most dangerous for bird's nests is Aspergillus fumigatus (Hubalek, 2000;Kornillowicz-Kowalska and Kitowski, 2013). The strains of this species cause fungal infections of the lungs and air sacs of birds (aspergillosis) (Barathidasan et al., 2013). Potentially pathogenic fungi isolated from the nests also includes keratinophilic fungi representing geophilic dermatophytes and related species considered as Chrysosporium ''group" (Kornillowicz-Kowalska and Bohacz, 2011). ...
Birds’ nests may be refuges for various species of fungi including that which are potentially phytopathogenic and zoopathogenic. Among the 2449 isolates of fungi obtained from nests of Marsh harriers 96.8% belonged to filamentous fungi. In total, 37 genera were identified from 63 fungi species. Within the mycobiotas of the examined nests populations of fungi which are potentially pathogenic for humans, homoiothermous animals and plants dominated. Among 63 species, 46 (72%) were potentially pathogenic fungi of which 18 species were potentially phytopathogenic and 32 species were pathogenic for homoiothermous animals. Inter alia species of fungi were found in the Marsh harriers nests: Aspergillus fumigatus, Aspergillus flavus, Scopulariopsis brevicaulis, Chrysosporium keratinophilum and Fusarium poae, Fusarium sporotrichioides. In terms of numbers, dominant in Marsh harrier nests were fungi pathogenic to birds, other homoiothermous animals and humans. On that basis it was concluded that Marsh harrier nests are both a source of fungal infections for that species and one of the links in the epidemiological cycle of opportunistic fungi for humans.
