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Review Article
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Fungal/Mycotic Diseases of Poultry-diagnosis, Treatment and Control: A Review |
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Kuldeep Dhama,
Sandip Chakraborty,
Amit Kumar Verma,
Ruchi Tiwari,
Rajamani Barathidasan,
Amit Kumar
and
Shambhu Dayal Singh
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ABSTRACT
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Fungal/mycotic diseases cause significant economic losses to the poultry industry either due to their direct infectious nature or due to production of mycotoxins, the secondary fungal metabolites produced in grains or poultry feed. Several fungi have created havoc in the poultry industry and some of them cause direct harm to human health due to their zoonotic implications. They are responsible for high morbidity and mortality, especially in young birds and cause stunted growth and diarrhea; and fatal encephalitis. Mycotic dermatitis is a possible health hazard associated with poultry houses. Mycotoxins are the leading cause of producing immunosuppression in birds, which makes them prone to several bacterial and viral infections leading to huge economic losses to the poultry industry. In comparison to bacterial and viral diseases, advances in diagnosis, treatment, prevention and control of fungal diseases in poultry has not taken much attention. Recently, molecular biological tools have been explored for rapid and accurate diagnosis of important fungal infections. Effective prevention and control measures include: appropriate hygiene, sanitation and disinfection, strict biosecurity programme and regular surveillance/monitoring of fungal infections as well as following judicious use of anti-fungal drugs. Precautionary measures during crop production, harvesting and storing and in feed mixing plants can help to check the fungal infections including health hazards of mycotoxins/mycotoxicosis. The present review describes the fungal pathogens causing diseases in poultry/birds, especially focusing to their diagnosis, prevention and control measures, which would help in formulating appropriate strategies to have a check and control on these unwanted troubles to the poultry producers/farmers.
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Received: January 09, 2013;
Accepted: March 03, 2013;
Published: May 16, 2013
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INTRODUCTION
Fungal/mycotic infections are common in all kinds of poultry birds but are
less prevalent as compared to bacterial and viral infections. Fungi are eukaryotic
organisms, comprising of both yeasts and molds. Fungal diseases of poultry include
Aspergillosis, Candidiasis, Dactylariosis, Cryptococcosis, Favus, Rhodotorulosis,
Torulopsis, Mucormycoses, Histoplasmosis and Cryptococcosis. Out of these, the
first two (Aspergillosis and Candidiasis) are having much importance and impact
and the last two (Histoplasmosis and Cryptococcosis) have some zoonotic significance.
Fungi produce disease in two ways viz. producing pathogenic signs and lesions
of disease by invading, harming and destroying body tissues of the host; and
by producing some toxins known as mycotoxins (aflatoxins, ochratoxins, ergot,
fusarium toxins etc.) in food grains and feed during crop production, harvesting
and storage steps, the intake, consumption and subsequent intoxication of which
produce disease, immunosuppressive condition and hampers production potential
(Hubalek, 1978; Jand and Singh,
1995; Jand et al., 2005; Kataria
et al., 2005; Dahlhausen, 2006; Dhama
et al., 2007, 2011a; Rai
et al., 2011; Singh et al., 2012).
Sporadic infections are common but sometimes they may take the form of outbreaks
(Kunkle, 2003a; Saif, 2003; Shivachandra
et al., 2004; Jand et al., 2005; Chauhan
and Roy, 2008; Dhama and Mahendran, 2008; Turner
et al., 2009; Dhama et al., 2011a;
Singh et al., 2012). Seasonal variation plays
important role in spread of fungal infections. Predominance of infection in
closed housing during summer and the presence of fungi in the poultry litter
material during autumn make the eradication difficult (Soliman
et al., 2009). Fungal diseases are assuming new importance because
of the inappropriate use of antibacterial that eliminate the natural beneficial
microflora which otherwise suppress the growth of fungi (De
Lucca, 2007). The fungal pathogens mainly target the respiratory and nervous
system of poultry and cause specific pathological changes in the host characterized
by inflammation, lesions and sickness leading to death (Shivachandra
et al., 2004). Chronic exposure to fungal spores produces allergic
responses in sensitized birds resulting in illness and decreased productivity.
Fungal infections require appropriate attention in terms of timely diagnosis
and effective treatment regimens to be followed. Advances in the treatment and
control of bacterial and viral diseases of poultry have been outstanding in
the recent years but the situation is not so good in case of fungal infections
and thus is a matter of concern (Jand and Singh, 1995;
Kunkle, 2003b; Dhama and Mahendran,
2008; Dhama et al., 2008, 2011a).
The discovery of mycophages (fungal viruses) have increased the expectation
as they can be exploited as a means of biological control and even to explain
the variation in antibiotic production and instability of fungal strains (Ghabrial,
1980). The present review describes the various fungal diseases of poultry
with emphasis on their diagnosis, treatment, prevention and control.
Aspergillosis (brooder's pneumonia): Aspergillosis, commonly known as
brooder's pneumonia, is caused mainly by Aspergillus fumigatus, most
pathogenic fungi affecting poultry (Arne et al., 2011)
but A. flavus has also been the culprit associated with many cases, Respiratory
infection by Aspergillus spp. has been reported in almost all
types of poultry birds viz., layer cockerels (Steinlage
et al., 2003), broilers (Martin et al.,
2007), growers (Zafra et al., 2008) and
turkey poults (Olias et al., 2010). Turkeys
are having higher susceptibility to aspergillosis when compared to chickens.
A. fumigatus infection occurs more frequently in poultry as the spores
of this pathogen species are smaller than those of other Aspergillus
spp. (Richard and Thurston, 1983; Arne
et al., 2011). Other Aspergillus spp. that may affect birds
adversely are A. terreus, A. glaucus, A. nidulans and A. niger
(Beernaert et al., 2010; Dhama
et al., 2012). Aspergilli can be isolated from environmental
samples and are worldwide in distribution. Spores of this fungal pathogen are
resistant in nature. Poultry birds coming in contact with the spores through
contaminated feed or litter gets affected after inhaling the spores. The predisposing
factors for flaring spore generation and dissemination in the air/environment
include warm environment, humidity, poor ventilation and sanitation along with
long term storage of feed (Tell, 2005; Khosravi
et al., 2008). The disease develops in brooder stages in chicks as
well as passerine birds, especially below three days of age (Pokras,
1988; Chauhan and Roy, 2008; McMillan
and Petrak, 1988). Exposure generally occurs by inhalation of spores, which
often originate from infected eggs that are opened. Chicks may get infection
in the hatcheries itself as by the release of large number of spores in the
environment and contaminate hatch mates (Oglesbee, 1997).
Aspergillosis is a necrotizing and granulomatous cavitory disease of the lungs
with hematogenous spread (Ganguly et al., 2011).
High humidity and moderate temperature conditions contributes significantly
towards the occurrence and spread of aspergillosis (Dhama
et al., 2008), thereby facilitating seasonal occurrence of the disease
in waterfowls with higher incidences in spring and autumn. Particularly, crippled
and malnourished captive birds suffer individually. Contaminant like lead acts
as a precipitating factor, especially in geese (Wobeser,
1997; Kapetanov et al., 2011).
Aspergillosis primarily causes high morbidity and mortality especially in young
chicks/birds (Redig, 2005; Arne
et al., 2011). The disease occurs in two main forms-acute and chronic.
Acute aspergillosis (brooder's pneumonia) occurs as a result of inhaling high
number of spores, wherein severe disease outbreaks in young birds are characteristically
observed. Morbidity and mortality are high (70-90%) in it and can be seen within
24-48 h of infection. Chronic form occurs sporadically and is the generally
observed in adult breeder birds (particularly turkeys) or occasionally in an
adult flock causing significant economic losses. This form is associated with
immune suppression (Vanderheyden, 1993). Proteases and
toxic secondary metabolites secreted by the fungus contribute to virulence (Tekaia
and Latge, 2005) along with gliotoxin, a highly immunosuppressive mycotoxin.
Air sacculitis is observed when concentrations of gliotoxins exceed 20- 70 μg
g-1 in poultry feedstuffs and in tissues of turkeys (Pena
et al., 2010). However, the true virulence factor is indecisive due
to environmental and clinical conditions inducing the disease condition in susceptible
birds. The pathogen clearance mechanisms in poultry rely on mucous-covered ciliated
epithelial cells lining the upper respiratory tract (Reese
et al., 2006). Lytic changes are observed in the epithelium of the
upper airway due to inhalation of conidia by even short duration of exposure
to contaminated environment (Nganpiep and Maina, 2002).
The initial physical barriers can be broken apart by the conidia of A. fumigatus
that are small enough (2-3 μm in diameter) to deeply penetrate the
respiratory system of birds.
Affected birds may show gasping along with fever, foetid diarrhea and rapid
loss of condition with convulsions occurring sometimes. Sub-acute form develops
within 8-10 days in birds upto 2 weeks of age with acute signs often present
in a milder form together with anemia. Respiratory rattle may be observed. Faeces
may also become yellowish (Reece et al., 1986;
Richard, 1997; Atasever and Gumussoy,
2004; Dhama et al., 2011a). Yellow coloured
pin point lesions are visible in lungs and air sacs, which can be seen through
naked eyes and may range from miliary to larger granulomatous foci. Sometimes
small yellow green granular fungus growth is observed in all the body cavities
with dry consistency of lungs. Walls of air sacs may thicken and bronchioles
may be filled with suppurates. Mycelial growth may extend into blood vessels
from where they disseminate. Granulomas can develop in multiple organs (Calnek
et al., 1997; Fraser et al., 1991; Shivaprasad,
2000). Distorted fruiting heads of the Aspergillus can be found in
air sacs. Turkeys suffer from chronic disease terminating in impedance of pulmonary
granulomas, causing right ventricular dilatation and ascites. Necrotic granulomatous
dermatitis and cutaneous aspergillosis are reported in chicken and pigeons,
respectively (Vanderheyden, 1993, Julian
and Goryo, 1990; Nardoni et al., 2006; Cacciuttolo
et al., 2009; Beernaert et al., 2010).
Large number of spores are found in wet litter and gets aerosolized when the
litter dries off. Biphasic mortality pattern is observed and in acute cases
mortality range between 5-50% during initial 1-3 weeks of age. Survivors often
develop chronic disease due to pulmonary insufficiency or neurological fungal
metastasis and may become lethargic and stunted (Calderone
and Fonzi, 2001; Beernaert et al., 2010).
Dyspnoea is common in neonates during first 3-5 days as evidenced by open mouth
breathing (gaspers) due to progressive airway obstruction. There may be affection
of eyes leading to deposition of cheesy materials (in turkeys) and blindness
along with Central Nervous System (CNS) abnormalities including torticollis
(Dyar et al., 1984; Jensen
et al., 1997; Throne Steinlage et al.,
2003; Dhama et al., 2011a).
Inhalation of large number of conidia from contaminated feed or litter affects
epithelium of conjunctiva and respiratory tract initiating granulomas which
causes pulmonary aspergillosis (Sauter et al., 1981;
Lugauskas et al., 2004). Opthalmitis occasionally
occur in turkeys upon experimental infection, wherein eyes may be cloudy with
retinitis, iridocyclitis and secondary involvement of the remainder of eye (Akan
et al., 2002). Necrotic foci in the cerebrum or cerebellum cause
encephalitis. Turkeys are quite sensitive to oral doses of necrotizing gliotoxin
(immunosuppressive and cytotoxic), produced by various isolates of A. fumigatus
and inhibit transformation of blood lymphocytes (Richard
et al., 1994, 1996; Richard
and DeBey, 1995; Peden and Rhoades, 1992). Birds
over 5 days of age show complete lesions along with uniform pinhead sized yellowish
nodules in lungs. Air sacs become thickened and cloudy having yellowish plaques.
Necrotic foci may occasionally be seen in the visceral organs (Okoye
et al, 1989; Steinlage et al., 2003;
Cacciuttolo et al., 2009; Singh
et al., 2009). Rapid death due to aspergillosis can flare up avian
influenza even though there is no connection between the two diseases and this
requires laboratory attention to distinguish these two diseases (Kradin
and Mark, 2008).
Diagnosis: Non-specific signs are common making diagnosis difficult
(Dahlhausen et al., 2004). Individual test does
not provide reliable diagnosis and therefore confirmatory diagnosis requires
disease history, clinical presentation, blood biochemical profile, serology,
radiographic changes along with endoscopy and cultural examination of the fungus
(Jones and Orosz, 2000). Stressful events are some adverse
environmental factors and/or an immunosuppressive condition or treatment (Jenkins,
1991). Chronic debilitation, voice change and exercise intolerance also
induce stress (Oglesbee, 1997). The clinical signs depend
on the form of the disease and involvement of organ (Jones
and Orosz, 2000), thereby requiring the disease to be differentiated from
other systemic diseases of respiratory tract (Jenkins 1991;
Jones and Orosz, 2000). Results of haematology and plasma
biochemistry are better diagnostic indicators (Jones and
Orosz, 2000). Serological tests include counter-immunoelectrophoresis, agar
gel immunodiffusion and enzyme-linked immunosorbent assays. However, negative
serological test results do not rule out aspergillosis; and positive tests must
be backed up by other disease evidences (Peden and Rhoades,
1992; Brown and Redig, 1994; Redig
et al., 1997; Le Loch et al., 2005;
Arca-Ruibal et al., 2006; Cray
et al., 2006, 2009a, b).
Although, radiographs may not be helpful, but lateral and ventrodorsal views
of a bird suspected for aspergillosis can give some indication and in absence
of anaesthesia, standing or perching lateral as well as dorsoventral views are
helpful (Jones and Orosz, 2000). Endoscopy of the abdominal
air sac can reveal a diffuse cloudiness or white or yellow plaques covered with
green gray pigmented mould. Samples for culture and cytology should be taken
directly with biopsy forceps or via air sac lavage (Jenkins
1991; Taylor, 1993; Oglesbee,
1997).
On necropsy, the granulomatous foci having varying degree of colour can be
noted in chronically ill patients (Jenkins 1991; Vanderheyden,
1993). Acute aspergillosis causes numerous miliary granulomatous foci (McMillan
and Petrak, 1989; Jenkins, 1991). Definitive diagnosis
requires demonstration of the organisms by cytology or histopathology and subsequent
identification by culture (Dahlhausen et al., 2004).
Isolation of the fungus alone does not confirm the infection status because
Aspergillus organisms are ubiquitous contaminants (Jensen
et al., 1997; Flammer and Orosz, 2008). However,
plentiful culturing from any organ should be considered for diagnosis, but a
negative culture also can not rule out Aspergillus infection (Redig,
2005; Jensen et al., 1997). Brain and heart
along with organs of respiratory system like larynx, trachea and lungs are important
for histopathological examination. Microscopic lesions can be suggestive but
not helpful in species identification because in vivo hyphae of hyaline
filamentous fungi are very similar and their in situ manifestations are
not pathognomonic (Kaufman et al., 1997; Tekaia
and Latge, 2005; Cray et al., 2009a). Thus,
immunohistochemistry usually can provide confirmatory diagnosis, although few
reports are only documented using monoclonal or polyclonal antibodies for diagnosing
aspergillosis in birds (Carrasco et al., 1993;
Jensen et al., 1997; Beytut
et al., 2004, Beytut, 2007).
Polymerase Chain Reaction (PCR) including real-time PCR assay is a valuable
diagnostic tool. PCR based cloning and sequencing of Internal Transcribed Spacer
(ITS) have been attempted successfully. Sophisticated techniques like Nucleic
Acid Sequence Based Amplification (NASBA) and Molecular Beacon (MB) technology
have increased the rapidity of diagnosis of this important pathogen (Calderone
and Fonzi, 2001; Dhama and Mahendran, 2008; Saleemi
et al., 2012; Dhama et al., 2011a,
2012; Zhao and Perlin, 2013).
Prevention and control: For prevention of aspergillosis, stress factors
and exposure to spores need to be minimized along with adopting strict hygiene
and sanitation measures in brooder and hatchery (Wright
et al., 1960; Chute and Richard, 1991, Beernaert
et al., 2010). Dirty, broken and potentially contaminated eggs must
be eliminated before setting in the incubator. An effective fungicide should
be applied inside the setter soon after transfer of hatching eggs is completed
(Wind and Yacowitz, 1960). Feed with low moisture content
should be given and the litter should be kept dry. Screened and elevated platforms
help to prevent turkeys from picking up molds from feed containers and waterer
fountains. Proper drainage is necessary to prevent water logging (Chute
and Richard, 1991). Maintain good ventilation, hygienic and stress-free
environmental conditions inside the poultry farm. A good litter management practice
needs to be followed and in between two flocks, treatment of new litter with
antifungal agent is mandatory to prevent the disease (Richard
et al., 1984; Shivachandra et al., 2004).
Feeders should be kept dry and clean to limit the fungal development (Powell
et al., 1994; Akan et al., 2002; Kunkle,
2003a). Affected and ill birds should be removed and culled/destroyed. Both
conventional and supportive treatment are required. In mild form of disease,
treatment is fruitful but when lesions are moderate to severe involving lungs
and air sacs, therapy is often not successful even after combination of drugs
are used. Various drugs like amphotericin-B, 5-fluorocytosine, ketoconazole
can be used to control the disease (Dhama et al.,
2012). Treating litter with Nystatin and Copper sulphate can reduce mold
content (Dyar et al., 1984). Copper sulphate
at 60 g quintal-1 of feed for 6 days is effective for treatment of
aspergillosis. In outbreaks, drinking water with 1:2000 aqueous solution of
copper sulphate needs to be provided. Tetracycline at 200 mg L-1
of drinking water should be given for 5 days to treat aspergillus infections
in chicks. Other drugs like eniconazole and fungicidin have also been tried
on experimental basis (Babras and Radhakrishnan, 1967;
Arne et al., 2011).
Candidiasis (moniliasis, thrush or sour crop): Candidiasis otherwise
known as thrush is a fungal disease caused by yeasts of the genus Candida
having nearly 200 species (Odds, 1994). Among them,
six are most frequently isolated. While C. albicans is the most abundant
and significant species, C. tropicalis, C. glabrata, C. parapsilosis,
C. krusei and C. lusitaniae have also been implicated as causative
agents. Susceptible hosts include domestic poultry, water fowls and wild birds
(Tiwari et al., 2011). Unhygienic atmosphere
and secondary debilitating conditions result in both superficial and deep infections.
Involvement of the digestive tract is common in young birds as compared to older
birds. Increased virulence of the fungus plays a vital role in establishing
the disease (Chute, 2001; Jungherr,
1933). C. albicans is an asporogenous and pseudomycelial dimorphic
yeast having fermentation capability. It grows on ordinary media over a wide
range of pH and temperature. Budding yeast forms (blastospores) are 3-4 μm
on epithelial surfaces whereas branching septate hyphae or pseudohyphae are
3-5 μm diameter in deeper tissues. It can utilize ammonia but not nitrate;
nitrogen and most strains need growth factor biotin to be supplemented for their
growth (Hubbard et al., 1986; Novak
et al., 2003). Endogenous form of the disease is most common due
to frequent presence of organisms in gastrointestinal tract and nature. C.
albicans is isolated from environmental sources less commonly than other
Candida species suggesting its adaptation to be parasitic, not saprophytic
way of life. Especially in psittacine birds, key entry and multiplication site
is the nasal cavity (Winner and Hurley, 1964; Balish
and Phillips, 1966; Tsai et al., 1992; Fulleringer
et al., 2006).
Transmission of Candida mainly occurs via fecal contaminated feed and water.
Candida spp. may become part of the inhabitant flora of the mouth, esophagus
and crop. Litter from poultry houses and game bird areas, waste and disposal
areas contaminated with human waste are suggested as potential sources for exposure
to Candida introduction (Bauck, 1994; Oglesby,
1997; Odds, 1988). Risk factors, which predisposes
to candidiasis and aggravate disease include malnutrition, vitamin D deficiency,
poor hygiene, prolonged use of antibiotic suppressing normal bacterial flora,
stress an immunosuppressive diseases (Campbell, 1986;
Kollias, 1986; Velasko, 2000)
Recognized virulent factors of C. albicans include adhesins having affinity
for the fibronectin on the cell surfaces, yeast forms cause tissue damage, phospholipase
concentrated in hyphal tips may enhance invasiveness, the mycelial phase of
C. albicans facilitates penetration of the fungus into tissues, cell
wall glycoprotein has an endotoxin like activity, (Meunier-Carpentier
et al., 1981, Ruchel et al., 1983;
Macdonald, 1984; Ruchel, 1984).
Neuraminidase, proteases, chitin, mannoprotein and lipids are other virulence
factors. Phenotypic switching in C. albicans may facilitate evasion of
host defense mechanisms. Systemic infection may occur when the fungus spread
via haematogenous route after vascular invasion by hyphae or pseudohyphae. Inflammatory
responses predominantly involve neutrophils and granulomatous lesions, but are
rare (Calderone and Fonzi, 2001; Schaller
et al., 2005; Antony et al., 2009).
Birds under 3 weeks of age are more susceptible to candidiasis. Affected poultry
show poor and stunted growth, depression diarrhea and dehydration which that
are altogether responsible for direct mortality (Chute, 1997;
O'Meara and Witter, 1971; Bauck, 1994).
Clinical signs are observed only in severely affected individual birds with
superficial oral or crop infections. In rare cases there is systemic invasion
wherein neurological, renal or intestinal involvements become evident (Chute,
1997; Oglesby, 1997; Velasko,
2000).
Lesions are usually confined to the upper digestive tract. Yeasts proliferate
on the surface and hyphae or pseudohyphae invade superficial epithelial layers,
causing hyperplasia and pseudomembrane or diphtheritic membrane formation, grossly
appearing as multifocal to confluent mats of cheesy material in the crop, but
less frequently in the esophagus and pharynx (Mayeda, 1961;
Velasko, 2000). Round raised ulcers or Turkish-towel
appearance in the mucosa are commonly observed (Bauck, 1994;
Schmidt et al., 2003). The membranous mass adhering
to the surface of the crop cannot be easily removed. Other areas of the upper
digestive tract develop false membranes resembling like that during diphtheria
and contain considerable necrotic tissue. Erosion of the lining of the proventriculus
and gizzard along with intestinal inflammation are commonly observed (O'Meara
and Witter, 1971; Odds, 1988; Chute,
1997; Bethea et al., 2010).
Diagnosis: For demonstrating hyphal forms of the yeast in the tissue,
diagnosis based on the lesions, histopathology and microscopic examination of
a digested smear are important. Cultural colonies of Candida appear as
white to ivory colour and smooth having a yeasty smell. C. albicans can
be isolated from faeces, crops, gizzards, lungs and livers. Isolation is done
by embryo inoculation test via chorioallantoic membrane (CAM). Fifty percent
of embryos may die between 48 and 72 h. Advent in molecular diagnostics generating
tools like PCR-RFLP has made the diagnosis easier and confirmatory (Ayatollahi
Mousavi et al., 2007; Tiwari et al.,
2011).
Treatment and control: As the organism has broad host range, cages,
equipments and other materials in contact with infected birds should be disinfected
without any delay. Cleanliness, adequate hygienic/disinfection measures, proper
managemental care and vitamin A supplementation are vital for disease prevention.
Excessive use of antibiotics and other stressors must be avoided (Bauck,
1994; Chute, 2001; Dhama et
al., 2003). Treatment of candidiasis should take care of the predisposing
conditions, risk factors or infections. Improved diet, husbandry and care can
minimise the severity of infection and subsequent lesions and losses. Addition
of vinegar to the drinking water acidifies the gastrointestinal contents, which
is unfavourable for fungal growth. Addition of chlorhexidine in the drinking
water helps to prevent overgrowth of Candida in poultry flocks or nurseries
(Underwood et al., 1956; Underwood,
1955; Smith, 1987). However, immune suppression
associated with the overuse of disinfectants need to be taken care of. Feeding
of birds with diets low in simple carbohydrates (i.e., grains and sprouts) is
advocated over seeds and sweets (Velasko, 2000). Invasive
and well established infections require the use of anti-fungal agents such as
Nystatin, azoles (fluconazole or itraconazole), or amphotericin B. For control,
nystatin (100 g ton-1) or copper sulphate (2-3 lbs ton-1)
to the feed for 7-10 days is prescribed (Flammer, 1993,
1994; Rupley, 1997; Velasko,
2000; Tiwari et al., 2011). Suboptimal management
conditions need to be avoided to prevent flaring up of the disease. Ideal practices
like continual use of mold inhibitors in the feed, proper feed storage and handling
practices, regular cleaning and sanitizing of the watering system and periodic
stirring and/or replacement of wet litter areas are essential elements for disease
prevention. Chlorine bleach added to the drinking water at 5 parts per million
(ppm) is quite effective (Janmaat and Morton, 2010).
Dactylariosis (mycotic encephalitis): Dactylariosis is caused by a dematiaceous
and thermophilic fungus-Dactylaria gallopava that affects young chicks
(Georg et al., 1964; Ranck
et al., 1974; Shane et al., 1985).
It grows well at 25-35°C with optimal temperature being 45°C and low
pH (<5) conditions. Spores spread after getting released into air (Tansey
and Brock, 1973; Waldrip et al., 1974; Randall
et al., 1981). Birds become infected on inhaling spores but the disease
is produced by angio-invasion and hematogenous spread to CNS. Birds between
initial 1-5 weeks of age are susceptible. Mortality during disease outbreak
ranges between 3-20%, mainly due to neurological disease (Blalock
et al., 1973; Waldrip et al., 1974;
Shane et al., 1985). Torticollis and in-coordination
due to paresis are observed in infected poults. Sometimes ocular lesions results
into blindness. In rare cases pulmonary granuloma causes dyspnoea as observed
in aspergillosis (Randall et al., 1981; Sonne
et al., 2012). Hematogenous spread of spores to brain leads to development
of lesions characterized by yellow or gray coloured meningeal or encephalitic
necrotic lesions that are more common in cerebellum or caudal cerebral cortex
(Blalock et al., 1973; Salkin
et al., 1990). Clinical signs and gross lesions are not specific,
making diagnosis difficult. Rapidly progressing nervous symptoms can also be
seen in young birds with vitamin E deficiency (encephalomalacia) or other bacterial
(meningitis) and viral (New castle disease) infections. Microscopically brain
lesions are reflected in pigmented 2 μm diameter hyphae and large number
of giant cells (Blalock et al., 1973; Ranck
et al, 1974). Saboraud Dextrose Agar (SDA) with suitable antibiotics
and incubation at 45°C is suitable for fungal isolation from brain samples.
Colonies produce brown color pigment diffusing into the surrounding medium and
have characteristic diploid conidia (Ranck et al.,
1974). Effective treatment does not exist for dactylariosis. So, avoiding
exposure to moldy litter, especially that with heat treatment is the only means
of prevention (Kunkle, 2003b).
Rhodotorulosis (mycotic dermatitis): Rhodotorulosis is caused by pink
yeast Rhodotorula, the yeast cells common contaminants and are infrequently
associated with disease conditions (Vazquez, 2011). The
fungus has been isolated from poultry litter and pigeon faecal droppings and
is of public health concern. R. glutinis produces dermatitis in broiler
chicken, while R. mucilaginosa cause dermatitis of feathers (Beemer
et al., 1970; Hubalek, 1978; Chauhan
and Roy, 1996; Alvarez-Perez et al., 2010).
This yeast predominantly associated with trachea of fowls and has even been
isolated from digestive organs (crop) along with Aspergillus fumigatus
and A. flavus. Birds die suddenly with crop highly distended and filled
with feed. Columbia agar with sheep blood (5%) or SDA with chloramphenicol supplementation
are ideal for Rhodotorula isolation. Fungal isolates can be identified
according to substrate accumulation profile and can be further confirmed by
skin biopsy (Page et al., 1980; Aruo,
1980; Grewal and Brar, 1987; Zaas
et al., 2003; Serena et al., 2004;
Tuon and Costa, 2008).
Favus (white comb): Favus is caused by Microsporum gallinae (Megnin)
(Trichophyton gallinae), Trichophyton simii, Microsporum gypseum
(Fonesca and Mendoza, 1984; Hubalek,
2000; Grunder et al., 2005). This
disease is not of much economical importance, occurs sporadically and as is
seen associated to demographic poverty. The fungal spores enter via unbroken
cutaneous surface during initial phase of infection, germinates in and around
the hair follicle and shaft (seldom) (Kane et al.,
1997). Lesions are observed on featherless skin areas like comb, wattle
and shanks; initially appearing as few grayish/yellowish cup like spots. They
increase in size and coalesce to make a wrinkled crust, which is mostly dry
and scaly appearing like honeycomb about the size of a pea. Feathered skin may
develop lesions of depression around follicles (favus cup), systemic signs are
not observed. Spread of infection occurs in birds by direct contact or via contaminated
fomites (Londero et al., 1969; Droual
et al., 1991; Saif, 2003). Favus is diagnosed
by demonstration of the fungi in the smears. Trichophyton gets easily
cultured on Saborauds glucose agar. Skin scrapings should be washed in
70% alcohol prior to attempting for cultural isolation (Bradley
et al., 1993; Saif, 2003). Microscopic examination
is performed with the skin scab examination on a glass slide with potassium
hydroxide solution (20%) and heated until appearance of a few bubbles; subsequently
it is examined for presence of fungi. Staining of the fungus can also be done
with 10% Parker Superchrome 51 pen ink in sodium hydroxide which demonstrates
the presence of fungus. Replacement of the birds with new stock need to be made
with disease free birds (symptom/lesion free). Proper segregation isolation
procedures need to be followed to avoid introducing the disease into a healthy
flock and to have check on its spread amongst the birds. If necessary, birds
should be culled and slaughtered. Dipping of the birds in 0.5% pentachlorophenol
or 5-bromosalicyl-4-chloranilide, a multi-fungine ointment, or Ayurvedic Himax
ointment (Indian Herbs Research and supply Co.) is useful in external application
(Chauhan and Roy, 2008).
Torulopsis infection: Torulopsis glabrata, the fungus
responsible for causing torulopsis in poultry, is a haploid and non-dimorphic
yeast. The disease is rarely seen in poultry and often is a problem in immunocompromised
birds. Pathogenicity of the fungal agent is dependent on the epithelial adhesion
genes, characteristically related with biofilm formation (Turner
et al., 2009). Liver gets enlarged in this disease condition and
reveals yellowish-white and well defined nodules of variable size. Round fungal
bodies with characteristic budding are observed in smear or section examination
on fungal staining. Clinical signs observed are dullness, loss of appetite,
ruffled feathers, etc. Identification is based on isolation of the causative
fungus but cultural isolation may take much time. Azoles viz. fluconazole, ketoconazole
etc are effective drugs of choice (Walker and Ayres, 1959;
Fidel et al., 1999; Chauhan
and Roy, 2008).
Mucormycosis: Chickens are less susceptible to mucor infection; pneumonic
lesions may be caused by Mucor resimosus or M. chorimbifer, while
few species may cause infection in the eyes and vertebrae (Migaki
et al., 1970; Chauhan and Roy, 2008). Mucor,
Penicillium and Aspergillus infections can occur through contaminated
litter. In advanced cases, there is frequent involvement of the sinuses, brain
and lungs. The infection can spread to gastrointestinal tract, skin and other
organs. The most common types of the disease conditions are oral and cerebral
mucormycosis (Spellberg et al., 2005; Auluck,
2007). Contaminated litter need to be removed to effectively control the
disease. Presumptive diagnosis requires biopsy examination of the affected tissue,
while examination of swabs of tissue or discharges is generally untrustworthy.
Administration of one table-spoonful of 33% potassium iodide solution in drinking
water per nearly 200 birds or antifungal drugs is helpful (Dawson
et al., 1976; Steinlage et al., 2003;
Dahlhausen, 2006).
Avian associated zoonotic fungal diseases
Cryptococcosis: Cryptococcosis, also known as Torulosis, Yeast meningitis,
Busse-Buschkes disease and European blastomycosis), is caused by Cryptococcus
neoformans that affects animals including poultry and humans (Singh
and Dash, 2008; Dhama et al., 2011b). Strains
differ in virulence and immune status of the host is crucial (Velasko,
2000). The incubation period is probably in weeks. Cryptococcosis is not
considered as true zoonoses. Fungus grows in soils with avian manures and thus
causes an indirect public health implication. Infections in birds are rare.
C. neoformans has been isolated from the feces of canaries (26%), carrier
pigeons (18%), budgerigars (2%) and psittacine birds (1%), apart from domestic
poultry (Saremi et al., 2004; Singh
and Dash, 2008). Humans acquire cryptococcosis from exposure to old pigeon
nests or droppings (Cafarchia et al., 2006;
Abdel-Razik, 2007; Rosario et
al., 2008). Transmission occurs via inhalation and occasionally by ingestion
and disease is usually chronic (Nosanchuk et al.,
2000; Malik et al., 2003; Rosario
et al., 2008; Dhama et al., 2011b).
Clinic signs in humans include meningitis pulmonary infection, dyspnoea, chest
pain, coughing, fever and malaise. Diarrhoea, weight loss, anaemia, headache,
paralysis, stiff neck and visual disturbances may occur (Velasko,
2000; Dhama et al., 2011b). Diagnosis requires
culturing the organism in SDA media, observing histopathological changes, but
with absence of inflammatory reaction. Cryptococci capsule develop a deep red
colour in Mucicarmine staining Specific treatment is not known and so the porognosis
is very grave (Mamidi et al., 2002; Nunez
et al., 2000). Necessary and appropriate cleanliness, disinfection,
hygiene and sanitation measures need to be followed to prevent the infection
and its zoonotic concerns (Aiello and Mays, 1998; King
and Markenday, 2004; Dhama et al., 2011b).
Histoplasmosis: Histoplasma capsulatum is a dimorphic fungus
commonly isolated from zoo birds and less frequently affects chicken and turkeys.
The soil enriched by decaying bird or bat droppings and litter with moisture
flourishes the growth of this fungi (Benenson, 1995;
Subramanian et al., 2005; Dhama
et al., 2011b). This disease is also not considered a true zoonotic
disease because the reservoir is soil and not the birds. Humans acquire infection
through inhalation of airborne spores. Histoplasmosis is not a contagious disease.
Humans are mostly affected by asymptomatic form, but acute pulmonary influenza-like
form can also be observed. The chronic form resembles tuberculosis and occurs
in people above 40 years of age. The disseminated form occurs in the very young
or the elderly and may have fatal consequences (Johnson
and Sarosi, 1987; Wheat et al., 1990; Mitchell,
1992; Dhama et al., 2011b). Patients should
be treated with an amphotericin B formulation and azole formulations (Dismukes
et al., 1992; Wheat et al., 1995;
Kaufman et al., 1997). The diagnosis requires
cultural identification of organism and histopathological examination revealing
extensive proliferation of reticuloendotlielial cells containing yeast forms.
Methenamine silver or periodic acid-schiff staining and histoplasmin sensitivity
testing are also useful for is detection. Conventional diagnostic tests like
Complement Fixation Test (CFT) and immunodiffusion along with those clinical
immunoassays that uses well characterized recombinant antigens (like Enzyme
immunoassays) have been given much emphasis. Recently, Polymerase Chain Reaction
(PCR) targeting the ribosomal RNA gene complex and real-time PCR targeting the
M specific protein are have been implied for diagnosis of H. capsulatum
(Guimaraes et al., 2006; Kauffman,
2006; Dhama et al., 2011b; Highland
et al., 2011). Biosecurity measures and appropriate precautions along
with following good hygiene and disinfection practices are necessary to prevent
the disease spread within birds and to prevent the transmission to humans (Mahvi,
1970; Dhama et al., 2003; Jacob
et al., 2011).
Discussion about fungal diseases remain incomplete without mentioning about
mycotoxins that appear in the food chain of both poultry and human as a result
of fungal infection of crops or poultry meat as they can resist degradation.
However, many international agencies are trying to achieve universal standardization
of regulatory limits for mycotoxins in poultry feed as well as products (Hussein
and Brasel, 2001). Moreover, the use of probiotics require a special mention
instead of relying on simply antifungals, as they can efficiently attack and
disrupt the fungal cell wall structures and poison their metabolic pathways,
thereby inhibiting their growth. If we consider the practical application of
probiotics, they significantly reduce the level of mycotoxins (Trias
et al., 2008). Advances in molecular diagnostic tools and techniques
along with novel emerging therapies need to be exploited to their full potential
for timely diagnosis and effective treatment of fungal diseases in poultry (Dhama
et al., 2010, 2013).
CONCLUSION AND FUTURE PERSPECTIVES
Prevention is better than cure
is the best policy to be adopted for fungal/mycotic disease problems in poultry.
No vaccine exist for any of the fungal diseases of poultry, therefore the timely
adoption of good management practices, strict biosecurity, effective disease
diagnosis and suitable preventive measures along with necessary treatment with
appropriate chemotherapeutic agents are the only elements to have a check and
control the fungal diseases of poultry. Apart from the fungal infections, mycotoxins
are also of major concern as they are the leading cause of immunosuppression
in birds, lowering their resistance level to various viral and bacterial diseases
and increased mortality. Thus, a holistic approach is required to combat the
adverse effects of mycotoxins and alleviate their adverse effects on high economic
returns from the poultry production. This requires regular surveillance and
monitoring of important mycotoxins with the use of conventional as well as modern
diagnostics. Emphasis is also need to be given for implementing suitable prevention
and control strategies for checking mycotoxin development and contamination
in feeds and their intoxication in poultry along judicious approach in the use
of probiotics.
|
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