Livestock Research for Rural Development 32 (4) 2020 LRRD Search LRRD Misssion Guide for preparation of papers LRRD Newsletter

Citation of this paper

Survey of aflatoxin B1 contamination in broiler feed from small-scale farms in Special Region of Yogyakarta, Indonesia

Muhsin Al Anas, Lies Mira Yusiati, Cuk Tri Noviandi and Ali Agus

Department of Animal Nutrition and Feed Science, Faculty of Animal Science, Universitas Gadjah Mada, Fauna 03, Bulaksumur, Yogyakarta, Indonesia 55821


This study aimed to determine aflatoxin B1 (AFB1) level contaminating broiler feed in Special Region of Yogyakarta, Indonesia. A total of 40 feed samples were collected from four districts in Special Region of Yogyakarta to measure AFB1 level by ELISA. The result showed that all feed samples in Special Region of Yogyakarta were contaminated by AFB1 approximately 169 g/kg. The lowest level of AFB1 contamination was in Sleman District, 79.59 g/kg. The level of AFB1 contamination in Kulon Progo District, Gunung Kidul District, and Sleman District did not differ;  AFB1 ranged from 332 to 474 g/kg. In conclusion, AFB1 contamination in the feed samples exceeded the Indonesian Standard (50 g/kg).

Keywords: Aspergillus, fungi, industry, mycotoxin, poultry


Aflatoxins are toxic secondary metabolites of Aspergillus flavus which contaminating many agricultural products and feeds. In the poultry industry, aflatoxin decreases productivity, increased mortality and impact on economic losses. Also, it caused immune suppression, liver damage, and metabolism inhibition (Zhang et al 2011; Kasmani et al 2012). Aflatoxin becomes a residue in the meat through the hydroxylated derivative, thus dangerous if consumed by humans (Hussain et al 2010). Aflatoxins are carcinogenic compound in humans and animals caused the ability of aflatoxins to bind deoxyribonucleic acid (DNA) (IARC 2002).

Tropical countries have a high potential for aflatoxins contamination in agriculture products (Daniel et al 2011; Probst et al 2014; Oliveira et al 2017). Indonesia is a tropical country that has an ideal environment for the growth of Aspergillus flavus (Ginting and Rahmianna 2015; Nurtjahja et al 2019). Besides, low levels of farmers' knowledge and awareness influence increasing in aflatoxin contamination (Udomkun et al 2018).

This study aimed to determine the aflatoxin contamination in broiler feed at small scale broiler farms in the Special Region of Yogyakarta. The results can be used to increase understanding and awareness related to aflatoxin contamination that is harmful to humans and animals.

Material and methods

Samples collection

This study was undertaken in four districts in Special Region of Yogyakarta (Figure 1). A total 40 broiler feed samples were collected for analyzing AFB1. The feeds ware collected from broiler farms to ensure feeds condition managed by farmers during the broiler period. Feed samples collected for analysis ranged from 1-2 kg. At the same time during farm visitation, the temperature and humidity were measured using a thermos-hygrometer (HTC-2 Thermometer Hygrometer). Districts geographical in Special Region of Yogyakarta are shown in Table 1.

Figure 1. Four districts of Special Region of Yogyakarta for feed sampling

Table 1. Districts geographical in Special Region of Yogyakarta (BPS-Statistics 2013)


Geographical Region


Latitude 7 34’ - 7 47’
longitude 110 33’ - 110 13’
Area 575 km2
Height 80 – 2,911 m.

Kulon progo

Latitude 70o 38’– 70o 59’
Longitude 110o 1’ – 110o 16’
Area 586 km2
Height 0 – 572 m.


Latitude 7 53’ – 17o 02’
Longitude 110 19' – 44o 00’
Area 507 km2
Height 0 – 80 m

Gunung Kidul

Latitude 7O 46’ - 8O 09’
Longitude 110o 21’ – 110o 50’
Area 1.485,36 km2
Height 150 – 700 m.

Laboratory analysis of aflatoxin level

Aflatoxin B1 contamination in poultry feed was analyzed by an Enzyme-linked immunosorbent assay (ELISA; Romer Labs Singapore Pte. Ltd.). A total of 1 kg samples were ground and kept in 2oC until analysis. Aflatoxin B1 was extracted from the feed following the manufacturer’s instruction by blending 50 g feed with 250 ml of 70% methanol. The sample was shaken or blend for 3 minutes. The supernatant was filtered through Whiteman No.1 filter paper. The absorbance of substrate samples was measured at 450 nm, using an ELISA reader.

Data analysis

Analysis of AFB1 contamination in broiler feed samples was compared with the maximum limit of AFB1 in broiler feed based on Indonesian National Standard (SNI), European Commission Recommendation (EU), and the Food and Drug Administration Regulatory Guidance for Toxins and Contamination of USA. Within SNI, the tolerance of AFB1 in broiler feed is 50 g/kg (starter and grower phases), 20 g/kg EU and USA standard. The difference in AFB1 contamination levels between districts in Special Region of Yogyakarta was analyzed with One-Way ANOVA.

Result and discussion

Temperature and humidity condition

The temperature and humidity in four districts in Special Region of Yogyakarta is shown in Table 2. The temperature and humidity did not differ in four districts. Indonesia is a tropical country with high temperature and humidity, thus supporting the growth of Aspergillus flavus in producing aflatoxin in many agricultural products (Noviandi et al 2001; Gruber-Dorninger et al 2019; Norlia et al 2019).

Table 2. Temperature dan humidity in four districts in Special Region of Yogyakarta


Temperature (oC)
(min and max)

Humidity (%)
(min and max)

Gunung Kidul









Kulon Progo



A. flavus could develop well in a proper environment to produce AFB1. Ideal temperature and humidity were main factors that would induce the enhancement of genes expression involved in the aflatoxin biosynthesis process (Abdel-Hadi et al 2011; Casquete et al 2017; Yunes et al 2019). Aflatoxin B1 can be produced optimally at a temperature of 20-30 oC and 90% humidity (Pertiwi et al 2015). The environment of Special Region of Yogykarta is perfect condition for Aspergillus growth in aflatoxins production.

Level of Aflatoxin B1 contamination in feed

The level of AFB1 contamination in feed on broiler farms in Special Region of Yogyakarta is presented in Table 3. Broiler feeds from several farms in Kulon Progo contained the highest AFB1 contamination level. However, these contaminations were not significantly different from Gunung Kidul and Bantul District. While the lowest AFB 1 contamination (p<0.05) was in Sleman District.

Table 3. Aflatoxin B1 contamination level (g/kg) in broiler feed in Special Region of Yogyakarta


of samples



95% Confidence Interval for Mean



Lower Bound

Upper Bound

Kulon Progo








Gunung Kidul
































a.b Different superscripts within a column indicate differences at p <0.05

The results of AFB1 analysis on feed samples in Special Region of Yogyakarta showed the level of AFB1 contamination exceeded the maximum level determined by SNI 50 g/kg (SNI 2009) and the United States standard (20 g/kg; Chen et al 2013). Contamination of AFB 1 in feed reaching levels above 100 g/kg caused a decrease in body weight of 7-17% (Denli et al 2009; Neeff et al 2013; Chen et al 2014), utilization of nutrients by livestock were not optimum, and mortality of poultry was increasing (Cravens et al 2013). Moreover, AFB1 consumed by poultry could be a residue in meat and liver (Hussain et al 2010; Fan et al 2013). These food products would be harmful to humans if it was consumed because aflatoxin was accumulated in the body and caused cancer (Bodreddigari et al 2008; Woo et al 2011).

The aflatoxin contamination in feed during the distribution chain (Yu 2012; Bai et al 2015) and storage process (harvest to market) would increase. Management of incompatible handling causes an increase in aflatoxin contamination to reach 5-200 times greater when it was through middlemen, transportation, city traders and market (market place) (Kamika et al 2016). Many feed warehouse conditions in farmers were not standard and high possibility of aflatoxin-contaminated (Figure 2).

Figure 2. Feed warehouse condition in small scale of broiler farms in Special Region of Yogyakarta. The feed was
placed in poor management of warehouse in high humidity without proper air circulation

Smallholder farmers were one of the factors causing increase aflatoxin contamination in feed. This was affected by the lack of knowledge and awareness about aflatoxin so that livestock management was not optimal. Aflatoxin contamination level would decrease if the implementation of distribution and storage management were handled well (Lee et al 2017; Marete et al 2019).

Collaborations for knowledge and technology dissemination between various parties including researchers, agricultural extension agents, stakeholders, government, and other farmers ware a method to prevent and reduce aflatoxin contamination level. The method could be started by applying a standard in the production chain sector involving seed systems and access, operational production, harvesting and processing, market operations, and utilization (Waliyar et al 2015, Udomkun et al 2017).



The authors would like to thank the Directorate-General for Science, Technology and Higher Education Resources, Ministry of Research and Technology/National Agency for Research and Innovation of the Republic of Indonesia for financial support of the research through Pendidikan Magister Menuju Doktor untuk Sarjana Unggul (PMDSU) scholarship.


Abdel-Hadi A, Schmidt-Heydt M, Parra R, Geisen R and Magan N 2012 A systems approach to model the relationship between aflatoxin gene cluster expression, environmental factors, growth and toxin production by Aspergillus flavus. Journal of The Royal Society Interface, 9: 757-767, Article Published April 7, 2012, from

Bai Y, Lan F, Yang W, Zhang F, Yang K, Li Z, Gao P and Wang S 2015 sRNA profiling in Aspergillus flavus reveals differentially expressed miRNA-like RNAs response to water activity and temperature. Fungal Genetics and Biology, 81: 113-119, Article Published Maret 24, 3015,

Bodreddigari S, Jones L K, Egner P A, Groopman J D, Sutter C H, Roebuck B D, Guengerich F P, Kensler T W and Sutter T R 2008 Protection against aflatoxin B1-induced cytotoxicity by expression of the cloned aflatoxin B1-aldehyde reductases rat AKR7A1 and human AKR7A3. Chemical Research in Toxicology 21: 1134–1142, Article Published November 24, 2013, from

BPS-Statistics 2013 Daerah Istimewa Yogyakarta in Figures 2013. BPS - Statistics of D.I. Yogyakarta Province. Yogyakarta. (in Bahasa Indonesia).

Casquete R, Benito M J, Cordoba M G, Ruiz-Moyano S and Martin A 2017 The growth and aflatoxin production of Aspergillus flavus strains on a cheese model system are influenced by physicochemical factors. Journal of Dairy Science, 100: 6987–6996, Article Published July 12, 2017, from

Chen X, Grenier B and Applegate T J 2013 Aflatoxins in poultry. Purdue Extention, AS-615W, Article Published February, 2013, from

Chen X, Horn N, Applegate T J 2014 Efficiency of hydrated sodium calcium aluminosilicate to ameliorate the adverse effects of graded levels of aflatoxin B1 in broiler chicks. Poultry Science 93: 2037-2047, Article Published June 3, 2014, from

Cravens R L, Goss G R, Chi F, De Boer E D, Davis S W, Hendrix S M, Richardson J A and Johnston S L 2013 The effects of necrotic enteritis, aflatoxin B1, and virginiamycin on growth performance, necrotic enteritis lesion scores, and mortality in young broilers. Poultry Science, 92:1997-2004, Article Published August, 2013, from

Daniel J H, Lewis L W, Redwood Y A, Kieszak S, Breiman R F, Flanders W D, Bell C, Mwihia J, Ogana G, Likimani S, Straetemans M and McGeehin M A 2011 Comprehensive assessment of maize aflatoxin levels in Eastern Kenya: 2005–2007. Environmental Health Perspectives, 119:1794–1799, Article Published August 15, 2011, from

Denli M, Blandon J C, Guynot M E, Salado S and Perez J F 2009 Effects of dietary AflaDetox on performance, serum biochemistry, histopathological changes, and aflatoxin residues in broilers exposed to aflatoxin B1. Poultry Science, 88(7): 1444–1451, Article Published July, 2019, from

Fan Y, Zhao L, Ma Q, Li X, Shi H, Zhou T, Zhang J and Ji C 2013 Effects of Bacillus subtilis ANSB060 on growth performance, meat quality and aflatoxin residues in broilers fed mouldy peanut meal naturally contaminated with aflatoxin. Food and Chemical Toxicology, 59: 748–753, Article Published July 17, 2013, from

Ginting E and Rahmianna A A 2015 Infection of Aspergillus flavus and physical quality of peanuts collected from farmers, local markets, and processors. Procedia Food Science, 3: 280–288, Article Published 2015, from

Gruber-Dorninger C, Jenkins T and Schatzmayr G 2019 Global mycotoxin occurrence in feed: a ten-year survey. Toxins, 11 (7): 375, Article Published June 27, 2019, from

Hussain Z, Khan M Z, Khan A, Javed I, Saleemi M K, Mahmood S and Asi M R 2010 Residues of aflatoxin B1 in broiler meat: Effect of age and dietary aflatoxin B1 levels. Food and Chemical Toxicology, 48: 3304-3307, Article Published Desember, 2010, from

IARC 2002 Some traditional herbal medicines, some mycotoxins, naphthalene and styrene. IARC Monogr Eval Carcinog Risks Hum, 82: 1–556. PMCID: PMC4781602

Kamika I, Ngbolua K and Tekere M 2016 Occurrence of aflatoxin contamination in maize throughout the supply chain in the Democratic Republic of Congo. Food Control, 69:292-296, Article Published November, 2016, from

Kasmani F B, Torshizi M A K, Allameh A and Shariatmadari F 2012 A novel aflatoxin-bindingBacillus probiotic: Performance, serum biochemistry, and immunological parameters in Japanese quail. Poultry Science, 91 (8): 1846-1853, Article Published August 1, 2012, from .

Lee H S, Nguyen-Viet H, Lindahl J, Thanh H M, Khanh T N, Hien L T T and Grace D 2017 A survey of aflatoxin B1 in maize and awareness of aflatoxins in Vietnam. World Mycotoxin Journal, 10 (2): 195-202, Article Published Januasry 16, 2017, from

Marete G N, Kanja L W, Mbaria J M, Okumu M O, Ateku P A, Korhonen H and Joutsjoki V 2019 Effects of the use of good agricultural practices on aflatoxin levels in maize grown in Nandi County, Kenya. Science, 1: 28, Article Published April 29, 2019, from

National Standardization Council 2009 National standard of maximum mycotoxin content in food (SNI 7385:2009) (In Bahasa Indonesia). Indonesian National Council for Standarization. Jakarta, 24 pp.

Neeff, D V, Ledoux D R, Rottinghaus G E, Bermudez A J, Dakovic A, Murarolli R A and Oliveira C A F 2013 In vitro and in vivo efficacy of a hydrated sodium calcium aluminosilicate to bind and reduce aflatoxin residues in tissues of broiler chicks fed aflatoxin B1. Poultry Science 92:131–137, Article Published January, 2013, from .

Norlia M, Jinap S, Nor-Khaizura M A R, Radu S, Samsudin N I P and Asilah A F 2019 Aspergillus section Flavi and aflatoxins: occurrence, detection, and identification in raw peanuts and peanut-based products along the supply chain. Frontiers in Microbiology, 10: 2602, Article Published November 22, 2019, from .

Noviandi C, Razzazi E, Agus A, Bhm J, Hulan H, Wedhastri S, Maryudhani Y, Nuryono, Sardjono and Leibetseder J 2001 Natural occurrence of aflatoxin B1 in some Indonesian food and feed products in Yogyakarta in year 1998-1999. Mycotoxin Research, 2:174-177, Article Published Juni 17, 2001, from

Nurtjahja K, Zuhra C F, Sembiring H, Bungsu A, Simanullang J, Silalahi J E, Gultom B N L and Sartini S 2019 Fungal contamination spices from Indonesia with emphasis on Aspergillus flavus. Czech Journal of Food Sciences, 37 (5): 338–344, Article Published 2019, from

Oliveira M O, Rocha A, Sulyok M, Krska R and Mallmann C A 2017 Natural mycotoxin contamination of maize (Zea mays L.) in the South Region of Brazil. Food Control, 73: 127-132, Article Published March, 2017, from

Pertiwi C, Rahayu W P, Lioe H N, Herawati D, Broto W and Ambarwati S 2015 The effect of temperature and relative humidity for Aspergillus flavus BIO 2237 growth and aflatoxin production on soybeans. International Food Research Journal, 22(1): 82-87.

Probst C, Bandyopadhyay R and Cotty P J 2014 Diversity of aflatoxin-producing fungi and their impact on food safety in sub-Saharan Africa. International Journal of Food Microbiology, 174: 113–122, Article Published March 17, 2014, from

Sugri I, Maalekuu B K, Gaveh E and Kusi F 2017 Sweet Potato Value Chain Analysis Reveals Opportunities for Increased Income and Food Security in Northern Ghana, Hindawi, Article Published November 2, 2017, from

Udomkun P, Wiredu A N, Nagle M, Muller J, Vanlauwe B and Bandyopadhyay R 2017 Innovative technologies to manage aflatoxins in foods and feeds and the profitability of application - A review. Food Control 76: 127-138, Article Published Juni, 2017, from

Udomkun P, Wossen T, Nabahungu N L, Mutegi C, Vanlauwe B and Bandyopadhyay R 2018 Incidence and farmers’ knowledge of aflatoxin contamination and control in Eastern Democratic Republic of Congo. Food Science & Nutrition, 6: 1607-1620, Article Published July 1, 2018, from

Waliyar F, Osiru M, Ntare B R, Kumar K V K, Sudini H, Traore A and Diarra B 2015 Post-harvest management of aflatoxin contamination in groundnut. World Mycotoxin Journal, 8 (2): 245-252, Article Published January, 2015, from

Woo L L, Egner P A, Belanger C L, Wattanawaraporn R, Trudel L J, Croy R G, Groopman J D, Essigmann J M and Wogan G N 2011 Aflatoxin B1-DNA adduct formation and mutagenicity in livers of neonatal male and female B6C3F1 mice leslie. Toxicological Sciences 122(1): 38–44, Article Published April 19, 2011, from

Yu J 2012 Current understanding on aflatoxin biosynthesis and future perspective in reducing aflatoxin contamination. Toxins, 4 (11): 1024-1057, Article Published October 25, 2012, from .

Yunes N B S, Oliveira R C, Reis T A, Baquiao A C, Rocha L O and Correa B 2019 Effect of temperature on growth, gene expression, and aflatoxin production by Aspergillus nomius isolated from Brazil nuts. Mycotoxin Research, Original Article Published December 11, 2019, from .

Zhang L, Ye Y, An Y, Tian Y, Wang Y and Tang H 2011 Systems responses of rats to aflatoxin b1 exposure revealed with metabonomic changes in multiple biological matrices. Journal of Proteome Research, 10 (2): 614-623, Article Published February 4, 2011, from

Received 2 February 2020; Accepted 28 February 2020; Published 1 April 2020

Go to top