Isolation and Identification of Some Opportunistic Fungi from Camels in Wasit Governorate of Iraq

The aim of this study was to investigate the percentage of opportunistic fungi and evaluate the diversity of yeasts and moulds associated with pulmonary diseases in camels in Wasit, Iraq. A total of 200 nasal cavity swabs (two swabs for each camel) were taken with sterile cotton swabs from 100 camels of different ages, sexes and areas. The results showed that 60 (60%) samples from 100 camels were positive for the occurrence of moulds and yeasts isolates, which classified into (16) species (37%) of moulds and (9) species (23%) of yeasts. In this study which revealed most frequent molds isolates were Asperigellius spp. specific A.fumigatus at percentage 7 (7%) followed by A.niger 5 (5%), 3(3%) for each A.flavus and A.terrus and Aspergillus spp 1(1%). On the other hand other molds Penicillum rubrum 3(3%), Penicillum spp.1(1%) followed by 2(2%) for Alternaria alternata ; Cladosporium; Mucor circinelloides; Mucor hiemalis and Mucor spp. and 1(1%) for Rizopus spp; Alternerria spp; Fusarium solani and Fusarium spp. were identifying. Also through this study shown the total isolation of yeasts were 23(%23) out of 100 camels revealed mainly frequent isolate were Candida spp. particular C.albicans at percentage 6 (6%) followed by 3(3%) for C.krusei; C. parapsilosis; C. tropicals and C. glabrata 1(1%). Other yeasts also can be identified such as Cryptococcus neoformans 2(2%); Geotricum candidum 2(2%) and 1(1%) for Malassezia spp. This study showed increases of moulds and yeasts isolation in camels with the increase age of animals. In conclusion: showed wide diversity of moulds and yeasts species isolated from camels, the most common molds isolate were Asperigellius spp. particular A.fumigatus while generally common yeasts isolate were Candida spp. specific C.albicans and increases of moulds and yeasts isolation in camels with the increase age of animals.

Fungi are eukaryotes organisms and everywhere in the environment, and can cooperate with plants, animals or humans, establish symbiotic, commensal, latent or pathogenic relationships 1 . Infection can be considered as an inequity between the host defenses and the infectious agent, with the host incapable to control the propagation of the contagious agent 2 . Fungal diseases will show if the immune system of the host is delicate 3 . The diagnosing is not easy since clinical beginning is diverse and depends on the host, treatment is difficult since number of available drugs is limited, and prevention is accessible for some fungi and only for some animal species 4 . Camels compared to other animals have been reported to be less subject to several diseases, thorough information on several aspects of the health status of camels are not well recognized. In spite of this, it has been reported that camels are subjected to various types microbial as well as fungal diseases 5,6 . Respiratory diseases in camels are consider to be one of the main rising troubles that lead to production loss and death 7 . However, only few studies are available on mycosis in the camels and their etiology in Iraq. Therefore, the aim of this study was to investigated the percentage of opportunistic fungi and evaluate the diversity of yeasts and moulds associated with pulmonary diseases in camels in Wasit, Iraq.

Study Area and Animals
A total of 200 nasal cavity swabs (two swabs for each camel) were taken with sterile cotton swabs from 100 camels of different ages, sexes and areas, during the period of November 2017 to April 2018. These areas include: Al-Mufqiya (40 camels), Al-Sheikh Saad (30 camels), Al-Bashair (20 camels) and villages of Hay city (10 camels) in Wasit province, Iraq.

Sampling and culturing
Two hundred samples of nasal cavity swabs have been collected from 100 camels. by sterile cotton swab, this swab were transferred to the laboratory of veterinary medicine, Wasit University for diagnosis after addition a small amount of sterile distilled water and these swabs directly were inoculated on Sabouraud dextrose agar and Cornmeal Agar plates with chlormphenicol, and incubated duplicated of culture at 30 °C and 37°C for two weeks 8 .

Molds Identification
Molds isolates were diagnosed according to cultural characteristics on Sabouraud dextrose agar, morphology of hyphae cells, spores and kind of fruiting bodies after staining with lacto phenol cotton blue 9 .

Yeast Identification
Yeasts isolates were diagnosed depending on cultural description on SDA that include color, shape and size to establish the morphology of the yeast cells. The following tests were used for the identification of the isolated yeast, Germ tube test 10 ; Dalmau plate technique on Cornmeal Agar 11 ; and API 20C AUX system (BioMerieux-France) were also performed according to the manufacturer's directions.

RESULTS
The study showed that 60 (60%) samples from 100 camels were positive for the occurrence of moulds and yeasts isolates, which classified into (16) species (37%) of mould and (9) species (23%) of yeast ( Table 1). The present study shown that incidence of fungal infection caused by moulds was more than yeasts those showed wide diversity of mould and yeast species. In this study which revealed most frequent molds isolates    Table 2). Also through this study shown the total isolation of yeasts were 23(%23) out of 100 camels revealed most frequent isolate were Candida spp. specific C.albicans at percentage 6 (6%) followed by 3(3%) for C.krusei; C. parapsilosis; C. tropicals and C. glabrata 1(1%). Other yeasts also can be identified such as Cryptococcus neoformans 2(2%), Geotricum candidum 2(2%) and 1(1%) for Malassezia spp. (Table 3). This study showed raise number of moulds and yeasts isolation in camels with the increase age of animals between 6 years to more than 10 years. While less isolation fungi were in age between 6 months to 3 years (Table  4).

DISCUSSION
The present study indicated a large range of fungal from nasal cavity swabs of camels were showed 60 (60%) out of 100 camels as positive for fungi isolation which included 37 (37%) of moulds and 23 (23%) of yeasts. These results corresponded with these mentioned by AL-Bashan and AL-Banki 12 who were able to isolate six different fungal species from nasal cavity swabs of Apparently Healthy Camels such as Aspergillus flavus, Aspergillus nidulans, Aspergillus niger, Aspergillus fumigatus, Penicillium sp. and Candida sp. Results of the current study is also in consistent with Gobrial et al. 13 who identification of moulds and yeasts fungi from nasal cavity swabs of the Egyptian camels it was found that Aspergillus sp.; Penicillium sp. and Candida sp. as well as other fungi. The current study found that most frequent molds isolates were Asperigellius spp.exacting A.fumigatus at percentage 7 (7%) compared with other Aspergillus sp. isolated was agreement with the finding of Lacey 14 found that a wide spread fungi around the world is genus Aspergillus and mainly pathogenic type causing disease in humans and animals its types; A. fumigatus belong to numerous factors; it's aptitude to grow more rapidly than other types in a broad range of temperature (20-50) ºC and it is extremely sporelating fungus. Other studies originate that Aspergillus fumigatus more common than other moulds when isolated fungi from other animals Ali and Khan 15 they found that the highest fungus related with abortion in cattle and buffaloes was A. fumigatus which has been recorded from over 60% of cases, also observations no scientific symptoms have been observed in the dam also before or after abortion.
The present study shown that incidence of fungal infection caused by moulds was more than yeasts. The nasal cavity as a part of respiratory system which contact immediately with outside system location led to simply entry of the spores moulds to the respiratory system by inhalation and accessible environment stander from temperature and humidity create from this system more contact to the fungi infection, it was supposed that housing of animals in relatively restricted spaces predispose them to infection due to the incidence of higher concentration of fungal spores in the air of cowsheds than that of its surrounding 15,16 . This was in agreement with previous report by Fekadu and Esayas 17 as it was mentioned above, pneumonia was among the mainly significant and frequently encountered disease of the camel. Although low mortality and morbidity rates, the improvement period was relatively long having negative contact on generally productivity.
In this study, it has been found that the total isolation of yeasts were 23(%23) out of 100 camels revealed most common isolate were Candida spp. particular C.albicans at percentage 6 (6%) and Other yeasts also can be identified such as Cryptococcus neoformans 2(2%); Geotricum candidum 2(2%) and1(1%) for Malassezia spp. The current result is in consistent with Osman et al. 18 who isolated Candida albicans from the nasopharyngeal cavity of apparently healthy camels at Shalateen, Halaieb and Abou-Ramad areas. suffering from different respiratory manifestations. Present results also agree with Al-Maadidhi 19 it was found C.albicans more common isolated from ewe of Iraq, this result in line with our result about C.albicans which was dominant isolated in nasal cavity swabs of camels. C. albicans was the species most commonly causes superficial and invasive infection were ability to adhere to diverse mucosa and epithelia, dimorphism, with assembly of pseudohyphae helping tissue invasion, thermotolerance, and exoenzymes like proteinase and phospholipase and germ tube configuration with subsequent advance of the filamentous form 20 . The mannan (glycoprotein present on the cell surface of C.albicans), adhesion responsible for the attachment of C.albicans to host cells more strong than other species of Candida 21 .
The current study found that increases fungal isolation in camels with the increase age of animals in age between 6 years to more than 10 years. While less isolation fungi were in age between 6 months to 3 years. Our result was in agreement with the finding of Wiserman et al. 22 they found that the percentage of fungal isolated increase with age of the animal. The current result also consistent with Al-Maadidhi 19 who studied the fungal infection in reproductive system of ewes, he establish that the percentage of fungal infection increase in the age animal. The causes possibly will due to the animals in this ages enhance facility to environment contact and also the animals are sexually active in this age which may be contamination through coating, parturition and abortion with other microorganism .

CONCLUSION
Showed wide diversity of moulds and yeasts species isolated from nasal cavity of camels, the most common molds isolate were Asperigellius spp. particular A.fumigatus while most common yeasts isolate were Candida spp. particular C.albicans and increases of moulds and yeasts isolation in camels with the increase age of animals.

ACKNOWLEDGEMENT
The author is grateful to all staff member of Microbiology Department College of Veterinary Medicine of Wasit University, for their help and cooperation.