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  1. Anjum A, Usman S, Aslam A, Faiz M, Usman S, Imran MS, et al.
    Trop Biomed, 2020 Jun 01;37(2):273-281.
    PMID: 33612797
    Contagious bovine pleuropneumonia (CBPP) is a highly contagious disease of cattle caused by Mycoplasma mycoides subsp. mycoides. It is characterized by anorexia, fever, dyspnea, polypnea, cough, and nasal discharges. Gross lesions in the lung such as marbling, sequestra, thickening of interlobular septa, and consolidation are evident. Serological tests including complement fixation test and competitive enzyme-linked immunosorbent assay and molecular tests such as polymerase chain reactions are used for diagnostic purposes. In this study, lung samples of suspected large ruminants (cattle n=560, buffalo n=293) were collected from abattoirs of three districts of Punjab namely Lahore, Kasur and Jhang. PCR was performed with specific primers, targeting the 16S ribosomal RNA gene to detect the positive cases. The results indicated that 49 samples (8.75%) of cattle were positive, with maximum prevalence was observed in Jhang with 16 positive samples (10.06%), but CBPP was not detected in any buffalo sample. High prevalence of disease was seen in cattle of more than seven years of age, in female cattle, and in cross-bred cattle. Age and gender were found significantly associated (P<0.05) with the prevalence of the disease. Gene sequencing of identified 5 isolates of Mycoplasma mycoides subsp. mycoides had more than 99% similarities with the strains isolated from China, Italy, Australia and Tanzania and were categorized into a monophyletic group but strain isolated from Portugal had more than 55% variable regions, hence clustered separately. This study confirms the presence of contagious bovine pleuropneumonia in the country which can be a threat to the livestock export market and warrants the implementation of control measures to mitigate the economic losses associated with the disease.
  2. Usman S, Razis AFA, Shaari K, Azmai MNA, Saad MZ, Isa NM, et al.
    Toxicol Rep, 2022;9:1369-1379.
    PMID: 36518379 DOI: 10.1016/j.toxrep.2022.05.001
    Microplastics (MPs) have become emerging pollutants of public health concern, due to their impact on aqua-terrestrial ecosystems and integration into the food web, with evidence of human exposure and unrevealed health implications. There is a paucity of information regarding the effects of MPs exposure on the gut system using metagenomic and metabolomic approaches. In this study, Javanese medaka fish was exposed to 5 µm beads of polystyrene microplastics (PS-MPs) suspensions, at concentrations of 100 μg/L (MP-LOW), 500 μg/L (MP-MED), and 1000 μg/L (MP-HIGH), for a duration of 21 days, and evaluated for gut microbiome and metabolome responses. The results revealed a significant reduction (p < 0.05) in richness and diversity of the gut microbiome in the MP-HIGH group, and identification of 7 bacterial genera as differential features by the Linear discriminant analysis Effect Size (LEfSe). The gut metabolic profile revealed upregulation of 9 metabolites related to energy metabolism, via tricarboxylic acid cycle (TCA), creatine pathway, and urea cycle, as determined by the pathway analysis. Furthermore, positive correlation was found between the genus Aeromonas and glucose, lactate, and creatine metabolites. The study revealed that PS-MPs exposure resulted in altered bacterial microbiome and metabolic disorder related to energy metabolism. It further provided additional data on gut bacterial genera and metabolites associated with MPs toxicity in aquatic organism, which will inevitably enable its future health risks assessment in animals and possibly humans.
  3. Iqbal SZ, Usman S, Razis AFA, Basheir Ali N, Saif T, Asi MR
    Int J Environ Res Public Health, 2020 Aug 03;17(15).
    PMID: 32756472 DOI: 10.3390/ijerph17155602
    The main goal of the present research was to explore the seasonal variation of deoxynivalenol (DON) in wheat, corn, and their products, collected during 2018-2019. Samples of 449 of wheat and products and 270 samples of corn and their products were examined using reverse-phase liquid chromatography with a UV detector. The findings of the present work showed that 104 (44.8%) samples of wheat and products from the summer season, and 91 (41.9%) samples from winter season were contaminated with DON (concentration limit of detections (LOD) to 2145 µg/kg and LOD to 2050 µg/kg), from summer and winter seasons, respectively. In corn and products, 87 (61.2%) samples from summer and 57 (44.5%) samples from winter season were polluted with DON with levels ranging from LOD to 2967 µg/kg and LOD to 2490 µg/kg, from the summer and winter season, respectively. The highest dietary intake of DON was determined in wheat flour 8.84 µg/kg body weight/day from the summer season, and 7.21 µg/kg body weight/day from the winter season. The findings of the work argued the need to implement stringent guidelines and create awareness among farmers, stakeholders, and traders of the harmful effect of DON. It is mostly observed that cereal crops are transported and stockpiled in jute bags, which may absorb moisture from the environment and produce favorable conditions for fungal growth. Therefore, these crops must store in polyethylene bags during transportation and storage, and moisture should be controlled. It is highly desirable to use those varieties that are more resistant to fungi attack. Humidity and moisture levels need to be controlled during storage and transportation.
  4. Aslam K, Iqbal SZ, Razis AFA, Usman S, Ali NB
    PMID: 33668973 DOI: 10.3390/ijerph18052270
    This research aims to assess the natural occurrence of patulin (PAT) in selected citrus fruits from central cities of Punjab and Pakistan's northern cities. A total of 2970 fruit samples from 12 citrus cultivars were examined using liquid chromatography fitted with a UV detector. The detection limit (LOD) and quantification limit were 0.04 and 0.12 µg/kg, respectively. About 56% of samples of citrus fruits from Punjab's central cities, Pakistan, were found to be contaminated with PAT, with values ranging from 0.12 to 1150 µg/kg in samples from central Punjab cities. Furthermore, 31.7% of samples of citrus fruits from northern cities of Pakistan were contaminated with PAT, with values ranging from 0.12 to 320 µg/kg. About 22.1% of citrus fruit samples had PAT levels greater than the suggested limits established by the European Union (EU). The dietary intake levels of PAT ranged from 0.10 to 1.11 µg/kg bw/day in the central cities of Punjab, Pakistan, and 0.13 to 1.93 µg/kg bw/day in the northern cities of Pakistan.
  5. Razis AFA, Shehzad MM, Usman S, Ali NB, Iqbal SZ, Naheed N, et al.
    PMID: 33276517 DOI: 10.3390/ijerph17238964
    A total of 779 samples of edible nuts (melon seeds, watermelon seeds, pumpkin seeds, and cantaloupe seeds) from Southern Punjab (Pakistan), were collected during the summer and the winter seasons. The natural occurrence of aflatoxins (AFs) and vitamin E (tocopherols) levels were investigated using HPLC. The results have shown that 180 (43.4%) of samples from the winter season and 122 (33.4%) samples from the summer season were found positive for AFs. Elevated average levels of total AFs (20.9 ± 3.10 μg/kg, dry weight) were observed in watermelon seeds without shell, and the lowest average amount (15.9 ± 3.60 μg/kg) were documented in melon seeds without shell samples from the winter season. An elevated average amount of total AFs 17.3 ± 1.50 μg/kg was found in pumpkin seeds available without a shell. The results have documented a significant difference in total AFs levels in edible seeds available with shells versus without shells (α = 0.05 & 0.01). The highest dietary intake of 6.30 μg/kg/day was found in female individuals from consuming pumpkin seeds (without shell) in the winter season. A value of 3.00 μg/kg/day was found in pumpkin seed without shell in the summer season in female individuals. The highest total tocopherol levels were 22.2 ± 7.70 ng/100 g in pumpkin seeds samples from the winter season and 14.5 ± 5.50 mg/100 g in melon seed samples from the summer season. The variation of total tocopherol levels in edible seeds among the winter and summer seasons showed a significant difference (p ≤ 0.0054), except watermelon seeds samples with non-significant differences (p ≥ 0.183).
  6. Waqas M, Iqbal SZ, Abdull Razis AF, Pervaiz W, Ahmad T, Usman S, et al.
    PMID: 34360308 DOI: 10.3390/ijerph18158015
    Aflatoxins (AFs) are secondary metabolites toxic to humans as well as animals. The environmental conditions, conventional agricultural practices, and illiteracy are the main factors which favor the production of AFs in food and feed. In the current study 744 samples of vegetable seeds and oils (soybean, sunflower, canola, olive, corn, and mustard) were collected and tested for the presence of aflatoxin B1 (AFB1) and total AFs. Liquid-liquid extraction was employed for the extraction of AFs from seeds and oil samples. Reverse phase high performance liquid chromatography equipped with fluorescence detection was used for the analysis. The results have shown that 92 (56.7%) samples of imported and 108 (57.0%) samples of local edible seeds were observed to be contaminated with AFs. All samples of edible seeds have AFB1 levels greater than the proposed limit set by the European Union (EU, 2 µg/kg) and 12 (7.40%) samples of imported seeds and 14 (7.40%) samples of local seeds were found in the range ≥ 50 µg/kg. About 78 (43.3%) samples of imported edible oil and 103 (48.3%) sample of local edible oil were observed to be positive for AFs. Furthermore, 16 (8.88%) and six (3.33%) samples of imported vegetable oil have levels of total AFs in a range (21-50 µg/kg) and greater than 50 µg/kg, respectively. The findings indicate significant differences in AFs levels between imported and local vegetable oil samples (t = 22.27 and p = 0.009) at α = 0.05 and a significant difference in AFs levels were found between vegetable seeds and oil samples (t = -17.75, p = 0.009) at α = 0.05. The highest dietary intake was found for a local sunflower oil sample (0.90 µg/kg/day) in female individuals (16-22 age group). The results have shown considerably high levels of AFB1 and total AFs in seeds and oil samples and emphasise the need to monitor carefully the levels of these toxic substances in food and feed on regular basis.
  7. Usman S, Abdull Razis AF, Shaari K, Amal MNA, Saad MZ, Mat Isa N, et al.
    PMID: 34574375 DOI: 10.3390/ijerph18189449
    Microplastics (MPs) have become pollutants of concern due to their unknown human health effect and negative impact on terrestrial and aquatic ecosystems. There is increasing number of experimental research on MPs globally with its effects not fully understood; recent animal studies explore its effects on the intestines, yet on other vital organs. Javanese medaka fish was exposed to polystyrene microplastics (PS-MPs) beads for a period of 21 days. Histological alterations, intestinal oxidative stress, permeability and neurotoxicity were evaluated. Significant inflammatory changes and tissue damage were observed in the intestine, liver and kidney. Intestinal oxidative stress and permeability were found to be significantly increased. In the brain, neurotoxicity characterised by a significant induction of oxidative stress, lipid peroxidation and the inhibition of acetylcholinesterase enzyme were elucidated. This study provided an insight into the multiple organ effect of microplastics exposure, necessitating further exploration and identification of biomarkers to be utilised for biomonitoring population at risk in the future.
  8. Usman S, Abdull Razis AF, Shaari K, Amal MNA, Saad MZ, Mat Isa N, et al.
    Int J Environ Res Public Health, 2020 Dec 21;17(24).
    PMID: 33371479 DOI: 10.3390/ijerph17249591
    Technological advances, coupled with increasing demands by consumers, have led to a drastic increase in plastic production. After serving their purposes, these plastics reach our water bodies as their destination and become ingested by aquatic organisms. This ubiquitous phenomenon has exposed humans to microplastics mostly through the consumption of sea food. This has led the World Health Organization (WHO) to make an urgent call for the assessment of environmental pollution due to microplastics and its effect on human health. This review summarizes studies between 1999 and 2020 in relation to microplastics in aquatic ecosystems and human food products, their potential toxic effects as elicited in animal studies, and policies on their use and disposal. There is a paucity of information on the toxicity mechanisms of microplastics in animal studies, and despite their documented presence in food products, no policy has been in place so far, to monitor and regulates microplastics in commercial foods meant for human consumption. Although there are policies and regulations with respect to plastics, these are only in a few countries and in most instances are not fully implemented due to socioeconomic reasons, so they do not address the problem across the entire life cycle of plastics from production to disposal. More animal research to elucidate pathways and early biomarkers of microplastic toxicity that can easily be detected in humans is needed. This is to create awareness and influence policies that will address this neglected threat to food safety and security.
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