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In the present study, fresh whole corn plants along with half milk kernels line were treated with live and hydrolysed yeast with different concentrations before ensiling and kept in airtight bags and then into mini silos in order to achieve anaerobic conditions for proper fermentation. The buckets were opened after different time intervals to characterise the material, quick acidification, dry matter recovery, and aerobic stability of silage respectively. Moreover, mycoflora and aflatoxin contamination were also analysed. The overall result reported that the silage quality was improved by the application of live and hydrolysed yeast. The best result was reported by the application of live yeast (T2: 10 g/kg) which significantly improved the fermentative, proximate, and digestibility parameters and reduced the mycoflora and aflatoxin contamination. Our results present promising new options for the use of natural compounds that may help to improve silage quality and reduce aflatoxin contamination. [ABSTRACT FROM AUTHOR]

Chickpea, Cicer arietinum L., is a nutrient rich crop that is widely cultivated and consumed in Pakistan. However, chickpea is highly prone to fungal growth leading to contamination with aflatoxins, the most potent carcinogen found in nature. In this study, fifty chickpea seed samples were collected from the local markets of the Punjab, Pakistan, to evaluate their nutritional quality, fungal and AFB1 contamination. Proximate analysis suggested that chickpea seeds contained 5.5–6.93% moisture, 62.24–63.24% carbohydrates, 22.75–23.44% protein, 4.99–5.4% fat, 5.62–5.84% fiber and 2.92–3.16% ash. Morphological identification techniques revealed fourteen fungal species belonging to six fungal genera from which Aspergillus flavus was the leading contaminant. AFB1 analysis revealed that sixty-two percent samples were contaminated with AFB1. All the AFB1 positive samples contained AFB1 level more than 2 ppb and 12.9% samples contain AFB1 level more than 20 ppb, exceeded the maximum limit (ML) assigned by EU and USA (FDA & FAO) respectively. The results of the present studies reported that chickpea is a highly contaminated commodity in terms of fungi and AFB1 that's why further investigations and monitoring are required to reduce the fungal and AFB1 contamination. These baseline data are an initial step in the effort to deal with this significant food safety issue. [ABSTRACT FROM AUTHOR]

Present research delves in the isolation, extraction and identification of mycotoxins from ten corn samples collected from the northern province of Pakistan. Average concentration of aflatoxin B1 and B2 by HP-TLC found in all corn samples was 27.87 and 1.35 μg/kg, respectively. Following HP-TLC, detoxification of the identified and isolated mycotoxin was performed, which was analyzed by HPLC. Screening of mycoflora exhibited Aspergillus niger and Fusarium as the most dominant fungal strains. Aflatoxin B1 was physically detoxified under UV-Lamp and direct sunlight displaying detoxification percentage of 48% and 99%, respectively. Biological detoxification involved the use of botanicals such as neem leaves, garlic and ginger powder, which portrayed an approximate detoxification of 70% from corn samples. Current research concludes that the tested physical and biological methods can be easily adopted at field and storage rooms after the harvesting of crops to avoid fungal contamination and subsequent food spoilage. [ABSTRACT FROM AUTHOR]

Maize (Zea mays L.) is considered as one of the main cereals, used as a source of food, forage, and processed products. The loss of maize productivity is reported due to effect on roots, stalks, ears, and kernels mainly caused by many fungi. Among these fungal pathogens of maize, Aspergillus flavus (A. flavus) are the most prevalent that produces highly toxigenic aflatoxins that are highly carcinogenic to the consumers. The present study is confined to isolate and characterize the A. flavus from maize seeds for accurate identification that can be helpful for determination and management of aflatoxins in maize crop. Eighty stored seed samples of maize were collected from warehouses where seeds are stored for food and feeding purposes. For the isolation of A. flavus, Potato Dextrose Agar was used. Isolated fungi were identified macro and microscopically using light microscope and scanning electron microscope. A total of 212 Aspergillus isolates were identified based on macro‐morphological and micro‐morphological characteristics. The results showed that A. flavus colonies were granular, flat with yellow‐green to deep yellow‐green colony color having a white border and compact, spherical spore heads. Rapid rate of growth was observed maturing in about 3–5 days. In microscopic features, A. flavus have apically swollen conidiophores with various conidia bearing cells in long and dry chains. Spherical conidial heads were split into several columns ranging 300–400 μm in diameter. This will be helpful for farmers, researchers and traders in future for correct identification of sources of aflatoxins. Research Highlights: Maize seed samples were collected from Pothohar region of Pakistan.The fungi were isolated on PDA.Aspergillus flavus was identified macro‐morphologically by observing growth rate, colony color and texture.Furthermore, these fungi were identified micro‐morphologically by using light and scanning electron microscope.The 212 Aspergillus flavus strains were isolated and identified. [ABSTRACT FROM AUTHOR]

Current research involves extraction, identification and detoxification of mycotoxins from ten dry fruit samples. Mycotoxins were identified by high performance thin layer chromatography followed by physical and biological detoxification, analysed by HPLC. Three fungal species were observed after isolation including, Aspergillus niger, Aspergillus flavus and Fussarium sp. HP-TLC analysis revealed the presence of mycotoxin, aflatoxin B1 ranging from 0.000303–0.03636 mg/kg in all samples. Results were further analysed through various statistical tests. Detoxification methods proved to be cost effective and easily implementable. Concentration of aflatoxin B1 in pine nuts was reduced to 0.0043 mg/kg and 0.0039 mg/kg in dry dates through UV based detoxification. Solarisation reduced the concentration of aflatoxin B1 in figs to 0.0044 mg/kg. 90% aflatoxins were detoxified by UV treatment while Zingiber officinale powder detoxified 90% mycotoxin. This research concludes that the studied detoxification methods can be generalised on larger scale to benefit the dry fruit industry worldwide. [ABSTRACT FROM AUTHOR]

This study was carried out to evaluate the prevalence of mycobiota and aflatoxin (AF) B1 contamination in sesame seeds from rainfed and irrigated zones of the Punjab, Pakistan. For this purpose, 100 sesame seed samples were collected directly from the fields of major sesame-producing areas in rainfed and irrigated zones. The agar plate method was used for isolation of mycobiota, and thin-layer chromatography was used to determine AFB1 concentrations. Seed samples were then stored for 12 months. After 12 months, the seeds were again analyzed for mycobiota and AFB1 for comparison. All samples were positive for fungal growth under fresh and stored conditions. Twenty-one fungal species of 10 genera were isolated. Aspergillus flavus was the most prevalent contaminant found in fresh and stored sesame seeds from rainfed and irrigated zones, followed by Aspergillus niger, Alternaria alternata, and Fusarium oxysporum. The least prevalent fungi were Aspergillus ochraceus and Cladosporium oxysporum. Analysis revealed that 92% of fresh and 99% of stored seed samples were contaminated with AFB1. In the rainfed zone, 88% of fresh and 100% of stored seed samples were contaminated with AFB1, with mean concentrations of 15.74 and 33.8 ppb, respectively. In the irrigated zone, 96% of fresh and 98% of stored seed samples were contaminated with AFB1, with mean concentrations of 20.5 and 27.56 ppb, respectively. AFB1 concentrations >20 ppb were found in 20% of fresh and 100% of stored seeds samples from the rainfed zone and in 28% of fresh and 60% of stored samples from the irrigated zone and thus were not fit for human consumption as per the maximum limit (20 ppb) assigned by the U.S. Food and Drug Administration and the Food and Agriculture Organization of the United Nations. This report is the first on the mycobiota and AFB1 contamination in sesame seeds from rainfed and irrigated zones of the Punjab, Pakistan. These baseline data are an initial step in the effort to deal with this significant food safety issue. Fresh and stored sesame seeds were positive for fungal growth. The leading contaminant in fresh and stored sesame seeds was Aspergillus flavus. Overall, 92% of fresh and 99% of stored seed samples were contaminated with AFB1. In the rainfed zone, 88% of fresh and 100% of stored samples were contaminated with AFB1. In the irrigated zone, 96% of fresh and 98% of stored samples were contaminated with AFB1. [ABSTRACT FROM AUTHOR]

Aflatoxin M1 (AFM1) is a hydroxylated metabolite of Aflatoxin B1 (AFB1). It appears in milk, when lactating animals consume AFB1 contaminated feed. It is carcinogenic and teratogenic in nature. Present study was planned to determine levels of AFM1 in raw and processed milk. For this, a total of five hundred and seventy milk samples (raw = 340 and processed = 230) were collected from Punjab (province of Pakistan). Processed milk included ultra-heat treated (UHT) (n=105), pasteurized (n=65), dried (n=40) and condensed milk (n=20). Concentration of AFM1 was quantified by direct competitive ELISA technique. Analysis revealed 100 percent incidence of AFM1 in UHT and pasteurized milk with a mean of 0.35±0.28ng/ml and 0.11+0.03ng/ml respectively. However, 86.66% raw milk samples were tainted with AFM1 with mean of 0.52±0.42ng/ml and 66.66% of dried milk samples with mean of 0.03+0.02ng/ml. However, none of the condensed milk sample was found positive. Data of raw milk contamination was further computed for seasonal variation. Highest prevalence (100%) was observed during autumn season followed by winter (81.81%), summer (80%) and spring season (62.06%) respectively. Furthermore, all mean values except raw milk were below the FDA legislation. Study results indicate the possible adverse effects on health of people of Pakistan. Good agriculture practices (GAP) and regular screening of raw materials of animal feed prior to supplying may help to control AFM1 levels in milk. [ABSTRACT FROM AUTHOR]

The present study evaluated Aspergillus (A) spp. and mycotoxins in fresh and ensiled maize fodder using a model fermentation system and raw materials from different areas of Punjab, Pakistan. A. niger, A. flavus and A. fumigatus were the most dominant followed by A. terrus and A. ochraceous in fresh and ensiled maize fodder. Total fungal counts ranged from 1×10³ to 4×10³cfu/ml for all species in both fresh and ensiled maize fodder. Total aflatoxins (AFB1, AFB2, AFG1 and AFG2) and ochratoxin A (OTA) analysis showed that AFB1was present with high frequency in fresh (37.5%) and ensiled (41.66%) fodder, with an average concentration of 9.49 and 8.36ng/g respectively. Aflatoxin B2 was detected in only two samples (16%) (1.2 and 1.3ng/g), aflatoxin G1 and G2 were not found. Ochratoxin A was found more frequently in fresh (54.16%) than in ensiled (20.86 %) samples with a mean of 8.06 and 4ng/g respectively. Average values for total aflatoxins (TAFs) and OTA were below the permissible limits as per regulation of European commission (EU), i.e. 20ng/g and 10ng/g, respectively. [ABSTRACT FROM AUTHOR]

This study presents the assessment of total aflatoxins (TAFs) in basmati rice (brown, 1,081; white, 1,170) collected from different areas of Punjab, Pakistan, during 2010 to 2015. Due to the carcinogenicity of TAFs, daily dietary exposure is also evaluated based on rice consumption survey data. Methodology was standardized by matrix spike recoveries at four fortification levels (0.1, 0.5, 2.5, and 12.5 ng/g) for TAFs (aflatoxins B1 [AFB1], B2 [AFB2], G1 [AFG1], and G2 [AFG2]). The present study reveals that 1,750 samples (77.74%) were tainted with AFB1, whereas TAFs were detected in 370 samples (16.43%). Of positive samples, 854 brown rice samples (79%) were positive for AFB1, and 154 samples (14.24%) were contaminated with TAFs. For white rice, 896 samples (76.58%) were contaminated with AFB1, whereas 205 samples (18.46%) were found positive for TAFs. Study findings were used to construct a frequency distribution, and AFB1 levels were also compared against permissible levels of TAFs (10 ng/g) as legislated by the European Commission. Results further revealed that daily dietary exposure of TAFs ranged from 0.51 to 10.22 ng/kg of body weight per day, which exceeds the permissible limit of 1 ng/kg of body weight per day as defined by the Joint FAO/WHO Expert Committee on Food Additives. [ABSTRACT FROM AUTHOR]

Eighty-four commercially available pet food samples including cat food (38 solid, 17 semi-solid) and dog food (20 solid, 9 semi-solid) were analyzed for melamine residues by competitive enzyme linked Immunosorbant assay. Among solid cat food, 78.94% (n=30) were contaminated with 14.43±1.27 (range >4-68 mg/kg) of melamine, while 82.35% samples of semi solid cat food (n=14) were found positive for melamine residue with mean value 6.43± 1.96 (range >2- 13.14mg/kg). Similarly, 75% samples of solid dog food (n=15) were positive for melamine residues with mean value 8.50±1.73 mg/kg (range >4-49mg/kg). A total of 44.44% samples of semi solid dog food were positive for melamine with mean value 16.31± 1.64 range of >2-34.25mg/kg. Out of all positive samples, 53% samples of pet food were contaminated beyond the Codex Alimentarius Commission (i.e. 2.5mg/kg). Analytical data showed existence of melamine residues in imported pet food. [ABSTRACT FROM AUTHOR]

Polymorphism exists in β-LG milk encoding gene in dairy animals which results in different protein isoforms being expressed m the milk. Overall quality of raw milk, total milk protein and fat content, depends on which β-LG isoforms are being expressed. Present study was planned to identify Beta-Lactoglobulin (β -LG) protein isoforms m Bubalm bubalis (Riverine buffalo) Nili Ravi breed milk and-to explore their association with major milk constituents. β-Lactoglobulin protein variants were typed using sodium dodecyl sulfate Polyacrylamide gel electrophoresis (SDS-PAGE). Total milk protein and whey contents were estimated using Bradford 1976 method while total milk fat by Gerber method. Three β-LG protein isoforms AA, BB and AB were identified with genotype frequencies; 0.49, 0.10 and 0.40 respectively. Allele A, with higher frequency of 0.69, was found more prevalent in studied animals as compared to B allele. The AA and AB isoforms were associated with high total milk protein contents (4.36%) while AB isoform had high whey protein (1.0%) and total milk fat (6.06%) contents Overall we report three β-LG protein isoforms (AA, AB and BB) in Nili Ravi buffalo milk which show significant association with major milk constituents. [ABSTRACT FROM AUTHOR]

The present study aimed to identify the presence of certain classes of phytochemicals in the leaf extract of medicinal herbs viz. Moringa oleifera and Calotropis procera , using qualitative detection tests and explored the potential of aqueous and ethanolic extract to inhibit aflatoxin production by thin layer chromatography at 25 °C and pH (7) of different incubation times i.e. 0-, 1-, 3-, 6- and 24-h. Qualitative phytochemical analysis reported that the aqueous leave extracts of M. oleifera and C. procera contained tannins, phlobatannins, quinones, steroids, sugar, betacyanins, fatty acids, phenols, and volatile oils. Aflatoxin analysis reported that the ethanolic extract of M. oleifera was found more effective than detoxifying 100% of AFB 1 after 24 h of incubation. In the case of C. procera , the aqueous extract reduced 96.5% of AFB 1 and ethanolic extract reduced 96% of AFB 1 after 24-h of incubation. The results revealed that natural plant products have a high potential to reduce AFB1 and could contribute to mitigation plans for AFB1. There is a need for further characterisation using techniques such as GC-MS, LC-MS, or NMR which would provide valuable information on the chemical composition of the extracts.

Chickpea, Cicer arietinum L., is a nutrient rich crop that is widely cultivated and consumed in Pakistan. However, chickpea is highly prone to fungal growth leading to contamination with aflatoxins, the most potent carcinogen found in nature. In this study, fifty chickpea seed samples were collected from the local markets of the Punjab, Pakistan, to evaluate their nutritional quality, fungal and AFB 1 contamination. Proximate analysis suggested that chickpea seeds contained 5.5-6.93% moisture, 62.24-63.24% carbohydrates, 22.75-23.44% protein, 4.99-5.4% fat, 5.62-5.84% fiber and 2.92-3.16% ash. Morphological identification techniques revealed fourteen fungal species belonging to six fungal genera from which Aspergillus flavus was the leading contaminant. AFB 1 analysis revealed that sixty-two percent samples were contaminated with AFB 1 . All the AFB 1 positive samples contained AFB 1 level more than 2 ppb and 12.9% samples contain AFB 1 level more than 20 ppb, exceeded the maximum limit (ML) assigned by EU and USA (FDA & FAO) respectively. The results of the present studies reported that chickpea is a highly contaminated commodity in terms of fungi and AFB 1 that's why further investigations and monitoring are required to reduce the fungal and AFB 1 contamination. These baseline data are an initial step in the effort to deal with this significant food safety issue.

Aflatoxin M1 (AFM 1 ) is a hydroxylated metabolite of Aflatoxin B 1 (AFB 1 ). It appears in milk, when lactating animals consume AFB 1 contaminated feed. It is carcinogenic and teratogenic in nature. Present study was planned to determine levels of AFM1 in raw and processed milk. For this, a total of five hundred and seventy milk samples (raw = 340 and processed = 230) were collected from Punjab (province of Pakistan). Processed milk included ultra-heat treated (UHT) (n=105), pasteurized (n=65), dried (n=40) and condensed milk (n=20). Concentration of AFM1 was quantified by direct competitive ELISA technique. Analysis revealed 100 percent incidence of AFM 1 in UHT and pasteurized milk with a mean of 0.35±0.28ng/ml and 0.11±0.03ng/ml respectively. However, 86.66% raw milk samples were tainted with AFM1 with mean of 0.52±0.42ng/ml and 66.66% of dried milk samples with mean of 0.03±0.02ng/ml. However, none of the condensed milk sample was found positive. Data of raw milk contamination was further computed for seasonal variation. Highest prevalence (100%) was observed during autumn season followed by winter (81.81%), summer (80%) and spring season (62.06%) respectively. Furthermore, all mean values except raw milk were below the FDA legislation. Study results indicate the possible adverse effects on health of people of Pakistan. Good agriculture practices (GAP) and regular screening of raw materials of animal feed prior to supplying may help to control AFM 1 levels in milk.