African Journal of Agricultural Science and Technology (AJAST) Vol. 2, Issue 4, pp.103-107. April, 2014 http://www.oceanicjournals.org/ajast ISSN 2311-5882 2014 Oceanic Journals Research Paper Study of fungi associated with farmers skin diseases in Sokoto Metropolis Shehu K 1, Muhammad S 1, Shinkafi SA 2, Salau IA 3* and Kasarawa AB 4 1 Department of Biological Sciences, Usmanu Danfodiyo University, Sokoto State, Nigeria. 2 Department of Microbiology, Usmanu Danfodiyo University, Sokoto State, Nigeria. 3 Department of Biology, Shehu Shagari College of Education, Sokoto State, Nigeria. 4 Department of Natural Sciences, Sokoto State Polytechnic, Sokoto State, Nigeria. *Corresponding author. E-mail: ibrahimasalau@yahoo.com or ibrahimasalau@gmail.com Tel: +234(0) 8053569551. Accepted 29 th April, 2014 A survey of fungi associated with skin diseases was carried out in three Local Government areas of Sokoto state (Wamakko, Sokoto North and Sokoto South) in 2012. Skin scrapped samples were collected from two hundred and ten (210) infected farmers using a multistage sampling technique. Mycological examination of the samples showed 66.3% prevalence of Dermatophytes. Most frequently isolated fungal pathogens from farmers skin were Trichophyton rubrum (38.4%), followed by Trichophyton mentagrophyte (25%) and Microsporum audouinii (20.8%). Microsporum gypseum was the least encounted dermatophyte with a prevalent rate of 15.5%. It is recommended that regular monitoring of the prevalence and distribution of Dermatophytes among farmers in Sokoto state and Nigeria in general can help strategise control options involving the use of chemical fungicides, wearing of protective gear such as rain boot and hand gloves and treatment of sewage prior to application in agricultural fields. Key words: Fungi, dermatophytes, farmers, Fadama and diseases. INTRODUCTION Fungi that cause skin diseases are generally referred to as dermatophytes (Agrios, 2005; Carlile et al., 2001). Many fungal species were associated with human skin diseases including ring worm, scalp, tinea, nails infection among others (Ashbee, 2000). The incidence of fungal disease has increased markedly over the last decade. Infections of skin, hair, nail and mucous membranes affect up to 20% of the population. Potentially fatal deep-seated infections have increased, mainly because of the larger numbers of people with a defective immune system (Stevens et al., 2003). Some fungal infections occur mainly in tropical or sub-tropical parts of the world and, in general, people from temperate zones, particularly Europe, have little immunity to them. Although, there is extensive literature on mycotic diseases affecting plants growing in fadama land as well as those found associated with human skin diseases from countries with farming practices similar to those being practice in Nigeria, the prevalence of these mycotic diseases of fadama plant cultivated in Nigeria and those associated with human skin diseases has not been extensively studied including those cultivated in Sokoto metropolis. Scanty information exists on the fungal pathogens associated with the fadama soil that has been responsible for vegetable and human skin diseases. The degree of pathological effects of diseases caused by the pathogens need be investigated in the study area. There is therefore, the need to isolate and identify the fungal pathogens and determine the prevalence of the diseases especially those associated with farmers in the Fadama land areas of Sokoto metropolis. About 76% of the dermatophytes species isolated
Afri. J. Agric. Sci. Technol. 104 are T. rubrum, 27% are T. mentagrophyte, 7% are T. verrucosum, 3% are T. tonsurans. Infrequently isolated (less than 1%) are E. floccosum, M. audouinii, M. canis,m. equinum, M. nanum, M. versicolor, T. equinum, T. kanei, T. raubitschekii, and T. violaceum (Demis, 1999). The present paper reports on the prevalence of fungi associated with farmers skin diseases in Sokoto, north- western Nigeria. MATERIALS AND METHODS Study of fungi associated with farmer s skin diseases was carried out in Sokoto, Sokoto is the capital city of the State, lies between latitude 13 3 490N, longitude 5 14 890E and at an altitude of 272 m above the sea level. It is located in the extreme North Western part of Sokoto North and South local government areas and also some parts of Wamakko LGA to the West. Sokoto metropolis is estimated to have a population of 427,760 people (NPC/FRN, 2007). Occupation of city inhabitants includes Farming, trading, commerce, with a reasonable proportion of the population working in private and public sectors (MOI, 2008). Sokoto is located in the Sudan savanna vegetation zone, where grass cover is more or less intersperse by short trees and shrubs. The soil is predominantly ferruginous tropical type, texturally sandy and ph of the soil ranges between 6 and 7. Rainfall starts in June and ends in September but may sometimes extend into October. The average annual rainfall is 550 mm with peak in the month of August. The highest temperatures of 45 C during the hot season are experienced in the months of March and April. Harmattan, a dry cold and dusty condition is experienced between the months of November and February (Udo and Mamman, 1993). Modern Sokoto city is a major commerce centre in leather crafts and agricultural products (MOI, 2008). In this study, a multistage sampling techniques was used where at the initial sampling, sixteen wards (61.5%) of the twenty six (26) wards within sokoto metropolis were selected. Then 60% of the total number of the wards selected, were randomly selected using systematic sampling method K= N/n (Thrushfied, 1995). From each selected area, clinically infected farmer with fungal skin disease were sampled. The sampling and laboratory analysis of the samples were completed within the period of six (6) months From May 2012 to October 2012. A total of two hundred and ten (210) samples of infected skin were collected from infected patients with clinical manifestation of dermatophytoses within Sokoto Metropolis (Fadama farms and Amanawa general hospital). The site of infection were first cleaned with methylated spirit, scales from the skin lesion were collected by scraping outwards with a blunt of scalpel from the edge of the lesion. Specimen from scalp was collected using forceps to pluck from the scalp. Samples were collected with the help of laboratory personnel from Specialist Hospital Sokoto and Amanawa General Hospital Dange-Shuni Local Government of Sokoto State. The samples were aseptically collected in paper envelop (5cm square). The samples were transported to Veterinary Microbiology Laboratory of the Faculty of Veterinary Medicine, Usmanu Danfodiyo University Sokoto for analysis. Samples collected and transported were processes within 6 h of collection. The media used were Sabouraud Dextrose Agar (SDA) and Potato Dextrose Agar (PDA), they were prepared according to the manufacturers instruction, Sixty two (62) g of SDA and forty two (42) g of PDA were weighed by using a digital weighing balance and dispersed into 1000 ml of distilled water into 1500ml conical flask, then plugged with cotton wool and wrapped with aluminium foil and then heated for complete dissolution. The dissolved media was then sterilized by autoclaving at 121 C for 15 min to ensure sterilization. The media was allowed to cool to 45 C and 1.3 ml of bacteria cell wall synthesis inhibitors (Gentamycin) was added to prevent bacterial contamination. This was then poured into sterilized Petri-dishes 15-20 ml, allowed to solidify for 24 h and sterilization, before inoculation of the samples. Skin scraped samples were inoculated on the surface of selected medium that is Sabouraud Dextrose Agar; the culture media were incubated at 30 C for up to 21 days. After the isolation the cultures were transferred to freshly prepared SDA to obtain the pure isolates. Species sporulate within seven (7) days, colors and conidia head were observed under the microscope the isolates were identified by their cultural morphology and microscopic characteristics. Identification was done to specie level by comparing their differentiating characteristics under microscope with diagrams in text book of mycological atlas (Chessbrough, 2003; Laura et al., 1998; Hartman and Rohde, 1980). Results were analyzed by three-factor analysis of variance with interactions. The P values and resulting conclusions were similar for all forms of analysis and P value = 0.05. Results were reported only for the untransformed data. Analyses by x 2 (using general linear model procedure) were performed using Minitab for Windows, release 14.2 (Minitab Inc., State College, PA). RESULTS A total of two hundred and ten (210) samples scraped human skins were collected from s farmers with symptoms of skin diseases. Of these, one hundred and sixty two (77.1%) were found to be positive for at least one species of fungal pathogens. Ninety 90 (42.9%) of
Salau et al. 105 the positive isolates were samples collected from infected males and seventy two 72 (34.3%) from females. The frequency of fungal skin pathogens isolation from infected males and females P<0.05 was presented in Table 1. In Sokoto South LGA. Tricophyton. rubrum (27) occurred more frequently with fourteen (14) in males and thirteen (13) in females. This was followed by T. mentagrophyte twenty five (25) with fourteen (14) in males and eleven (11) in females, M. gypseum eighteen (18) with nine (9) in males and nine (9) in females, A. niger fourteen (14) with six (6) in male and eight (8) in female, A. flavus has ten (10) with six (6) in males and four (4) in females. Occurrence of the predominant fungal pathogens isolated was less differences in males than females. The Chi-square results in Table 3 shows that at P>0.05, there was no significant difference between males and females. The least in occurrences were isolates of Aspergillus fumigatus six (6) with four (4) in males and two (2) in Table 1. Prevalence of fungal infection on human skin. Source +ve for pathogens - ve for pathogens Total Male 90 15 105 Female 72 33 105 Total 77.1% 48 (22.9%) 210 Chi-Sq = 8.750, DF = 1, P-value = 0.003. Table 2. Fungi associated with human skin infections in Sokoto South LGA. Isolates Sample from Male (n=53) Sample from Female (n=52) Total (n=105) A. niger 06 08 14 A. flavus 06 04 10 A. fumigatus 04 02 06 T. rubrum 14 13 27 T. mentagrophyte 14 11 25 M. gypseum 09 09 18 M. audouinii 00 05 05 Chi-Sq = 6.741, DF = 6, P-Value = 0.346. 5 cells with expected counts less than 5. Table 3. Fungi isolated from infected human skin in Sokoto North LGA. Isolates Sample from Male (n=76) Sample from Female (n=50) Total (n=126) A. niger 19 12 31 A. flavus 07 05 12 A. fumigatus 02 08 10 T. rubrum 18 07 25 T. mentagrophyte 07 05 12 M. gypseum 06 03 09 M. audouinii 17 10 27 Chi-Sq = 8.499, DF = 6, P-Value = 0.204. 4 cells with expected counts less than 5. Table 4. Fungi isolated from infected human skin in Wamakko LGA. Isolates Sample from Male (n=74) Sample from Female (n=59) Total (n=133) A. niger 11 08 19 A. flavus 08 O6 14 A. fumigatus 04 04 08 T. rubrum 26 14 40 T. mentagrophyte 13 10 23 M. gypseum 05 06 11 M. audouinii 07 11 18 Chi-Sq = 4.091, DF = 6, P-Value = 0.664. 3 cells with expected counts less than 5. females, and Microsporum audouinii with five (5) isolates all in females. The frequencies of various types of fungal pathogens isolated from Sokoto South ward are presented in Table 2.
Afri. J. Agric. Sci. Technol. 106 This was followed by M. audouinii (27) with seventeen (17) in males and ten (10) in females, T. rubrum (25) with eighteen (18) in males and seven (7) in females, then A. flavus and T. mentagrophyte (12) each with seven (7) each in males and five (5) in females. Occurrence of the predominant fungal pathogens isolated was more in males than females P>0.05. The least in occurrences were isolates of Aspergillus fumigatus and Microsporum gypseum ten (10) and nine (9) isolates respectively. The frequency of various types of fungal pathogens isolated from humans in sokoto north is presented in Table 3. In Wamakko, Trichophyton rubrum (40) occurred more frequently with twenty six (26) in male and fourteen (14) in female. This was followed by Trichophyton mentagrophyte (23) with thirteen (13) in male and ten (10) in female, Aspergillus niger (19) with eleven (11) in male and eight (8) in female, Microsporum audouinii eighteen (18) with seven (7) in male and eleven (11) in female, Aspergillus flavus fourteen (14) with eight (8) in males and six (6) in females. Occurrences of the predominant fungal pathogens isolated were more in males than females P>0.05. The least in occurrences were isolates of Microsporum gypseum and Aspergillus fumigatus eleven (11) and eight (8) isolates respectively. The frequency of various types of fungal pathogens isolated from humans in Wamakko wards is presented in Table 4. DISCUSSION Results obtained from this study, indicate that seven (7) fungal pathogens were isolated they include: A. niger, A. fumigatus, A. flavus, T. rubrum, T. mentagrophyte, M. Gypseum and M. audouinii, were found to be the causes of human mycotic skin infection. Is in agreement with the findings of Mahmoudabadi (2008) who reported T. rubrum, T. mentagrophyte, M. Gypseum and M. audouinii as fungal pathogens associated with human skin diseases. The finding was also in conformity with the report of Hasegawa (2000) who reported species of dermatophytes and candidas are responsible for human skin diseases. The frequency of occurrence of these species indicated that Dermatophytes have the highest percentage of prevalence of (77.1%) this agreed with the findings of Summerbell (1997) where dermatophytes account for over 90% of skin toenail infections and the majority of fingernail infections. The most commonly isolated species in Europe and North America as reported by Blake et al. (1991) are T. rubrum and T. mentagrophyte and Epidermophyton floccosum as less predominant specie. Similarly in this study T. rubrum occurred more frequently than other dermatophytes followed by Trichophyton mentagrophyte and Microsporum audouinii then Microsporum gypseum. This agree with the findings of Hartman and Rohde (1980) who reported T. rubrum as the most common pathogenic fungi of man and counts for 58% of the dermatophytes species isolated Shinkafi (2011). Also is in agreement with the report in Ellis et al. (1997) who isolated similar pathogens as the most common dermatophytes worldwide responsible for human skin diseases. According to Elewski and Hazen (1989), and Rippon (1988), people in the endemic areas do not wear footcover and the dermatophyte species contact with the exposed skin can invade and cause tinea corporis Worldwide, the anthropophilic species. The next organisms in terms of percentage prevalence is T. mentagrophyte according to laura et al. (1980) about 28% percentage of the Dermatophytes species isolated is T. mentagrophyte. 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