Mothers' Knowledge, Attitudes, and Practices Regarding the Prevention and Management of Diarrhea among Children in Pakistan: A Cross-Sectional Study.

Diarrheal disease is the second leading cause of death worldwide in children under 5 years old, after pneumonia. Fortunately, diarrhea is a preventable disease that can be avoided by implementing basic home management strategies. Mothers are essential to its management and prevention; therefore, this study assessed the knowledge, attitudes, and practices of mothers in Pakistan related to diarrheal disease prevention and management. The study was conducted using a cross-sectional design in three cities of Pakistan from September 2022 to December 2022. A questionnaire was used to collect data on mothers' sociodemographic characteristics and their knowledge, attitudes, and practices related to the prevention and management of diarrheal diseases. A total of 356 mothers (81.7% of them were housewives, and 58.4% were 25-34 years old) participated in the study. Data were analyzed using descriptive statistics and tests of association. Significant associations were found between mothers' income, education, and ethnicity and their knowledge, attitudes, and practices regarding the prevention and management of diarrheal diseases (P <0.05). However, no significant association was found between the other variables. The knowledge and attitudes of the mothers regarding the prevention and management of diarrhea were satisfactory; however, their prevention-related practices and home-based management were unsatisfactory. Therefore, community education, formation of health and hygiene committees, and dissemination of user-friendly information are crucial for creating awareness about the prevention and management of diarrheal diseases. These measures can help improve the practices of mothers and reduce the incidence of diarrheal diseases in Pakistan.

Complications of Therapeutic Plasma Exchange in pediatric patients: An experience at a tertiary care hospital.

Objective: To find the incidence of various complications of therapeutic plasma exchange (TPE) in ICU admitted children and to determine their association with age, gender, blood group and diagnosis of the patients. Methods: In this observational study, data of 24 patients who underwent 125 sessions of TPE was collected from the Pediatric Intensive care unit (PICU) and Hematology department of The Children's Hospital, Lahore from December 2020 to November 2021. Age, gender, blood group, indications and complications observed during and after the TPE procedure were documented on a pre-designed proforma. The data was analyzed by using SPSS version 23. Quantitative variables were presented in the form of mean and standard deviation. Qualitative variables like gender, blood groups, indications and complications of plasmapheresis were presented as frequency and percentage. Chi square test was applied for comparison of variables. Results: Among the 24 patients, 45.8% were of age group five to ten years with mean age of 7.58 years± 2.04 years and male to female ratio of 0.84:1. Guillain-Barré syndrome (GBS) and Neuromyelitis Optica spectrum disorder (NMO-SD) were the most prevalent among the patients who underwent TPE. Most common complication was hypotension (44.9%), others were febrile reactions (11.6%), unstable vital signs (14.5%) and allergic reactions (24.6%). Blood group, clinical condition and diagnosis of the patient showed significant association with the incidence of TPE related complications. Conclusion: The majority of problems caused by TPE are considered to be minor. Sudden fall in blood pressure, pruritus, urticarial rash and fever are the common adverse consequences among pediatric patients. Blood group and diagnosis of the patient can determine the development of such complications during plasmapheresis procedure.

The prevalence, risk factors analysis and evaluation of two diagnostic techniques for the detection of Cryptosporidium infection in diarrheic sheep from Pakistan.

BACKGROUND: Cryptosporidium spp. is recognized as an opportunistic zoonotic parasite that infects humans as well as wild and domestic animals. This enteric protozoan is a major cause of diarrhea in humans and animals and often result in death due to severe dehydration. The present study was designed to investigate the prevalence, identification of various risk factors and evaluation of sensitivity of the two diagnostic techniques for rapid and correct detection of Cryptosporidium infection in diarrheic sheep in Pakistan. METHODS: A total of 360 fecal samples were collected and processed for detection of Cryptosporidium infection after proper preservation. These samples were properly stained with modified Ziehl-Neelsen acid staining and then examined under simple microscope at 100x magnification for confirmation of Cryptosporidium oocysts. The same samples were again processed through simple PCR for confirmation of the Cryptosporidium spp. RESULTS: The age wise prevalence was detected through simple microscopy and PCR. We found highest prevalence at the age of ≤1 year followed by 1-2 years of age while the lowest prevalence was recorded at the age of ≥ 2-3 years of sheep and found significant difference between different ages (P<0.05). The sex wise prevalence showed the highest prevalence in male (♂) animals detected compared to female (♀). The overall prevalence was detected 27.08% and 18.80% through PCR and simple microscopy, respectively, and significant difference between two diagnostic techniques were observed (P<0.05). Considering the seasonality, the highest prevalence was recorded through simple microscopy in autumn, summer, and spring, while the lowest in winter. These results were confirmed through PCR. CONCLUSION: It was concluded that molecular detection is the most efficient, specific and sensitive technique for detection of Cryptosporidium infection than simple microscopy. Moreover sheep is the major potential source of infection to other wild and domestic animals including humans.

Association study of six candidate genes with major depressive disorder in the North-Western population of Pakistan.

People around the world are currently affected by Major Depressive Disorder (MDD). Despite its many aspects, symptoms, manifestations and impacts, efforts have been made to identify the root causes of the disorder. In particular, genetic studies have concentrated on identifying candidate genes for MDD and exploring associations between these genes and some specific group of individuals. The aim of this research was to find out the association between single nucleotide polymorphisms in 6 candidate genes linked to the neurobiology of major depressive disorder in the North-Western population of Pakistan. We performed a case-control analysis, with 400 MDD and 232 controls. A trained psychiatrist or clinical psychologists evaluated the patients. Six polymorphisms were genotyped and tested for allele and genotype association with MDD. There were no statistical variations between MDD patients and healthy controls for genotypic and allelic distribution of all the polymorphisms observed. Thus, our analysis does not support the major role of these polymorphisms in contributing to MDD susceptibility, although it does not preclude minor impact. The statistically significant correlation between six polymorphisms and major depressive disorder in the studied population was not observed. There are inconsistencies in investigations around the world. Future research, including GWAS and association analysis on larger scale should be addressed for further validation and replication of the present findings.

Evaluation of biological and chemical additives on microbial community, fermentation characteristics, aerobic stability, and in vitro gas production of SuMu No. 2 elephant grass.

BACKGROUND: The study was conducted to evaluate the effects of biological and chemical additives on microbial community, fermentation characteristics, aerobic stability, and in vitro gas production of SuMu No. 2 elephant grass. RESULTS: Aerobic bacteria and yeast were not affected on days 5 and 7 but were significantly (P < 0.224) reduced on days 14, 30, and 60, whereas lactic acid and lactic acid bacteria were significantly (P > 0.001) higher in all ensiling days within all treatment groups. During the ensiling days, the pH, acetic acid, butyric acid, and yeast were decreased in all treatment groups, whereas the Lactobacillus plantarum group and L. plantarum + natamycin group were highly significantly (P > 0.001) decreased. During air exposure, the water-soluble carbohydrates, ammonia nitrogen, lactic acid, and acetic acid were not affected on days 1-4, whereas pH and aerobic bacteria (were significantly (P < 0.05) increased on days 2-4. The addition of Lactobacillus plantarum and natamycin increased the gas production, in vitro dry matter digestibility, and in vitro neutral detergent fiber of SuMu No. 2 elephant grass silages. CONCLUSIONS: The addition of biological and chemical additives, such as L. plantrum alone and the combination with natamycin, affected the undesirable microbial community, fermentation characteristics, aerobic stability, and in vitro gas of SuMu No. 2 elephant grass. © 2021 Society of Chemical Industry.

Antidiabetic and Hypolipidemic Potential of Green AgNPs against Diabetic Mice.

Green nanotechnology-based approaches have been acquired as environmentally friendly and cost effective with many biomedical applications. The present study reports the synthesis of silver nanoparticles (AgNPs) from the leaves of Emblica phyllanthus, characterized by UV-Vis spectroscopy, EDX, SEM, AFM, and XRD. The acute and chronic antidiabetic and hypolipidemic potential of AgNPs was studied in alloxan-induced diabetic mice. A total of 11 groups (G1-G11, n = 6) of mice were treated with different concentrations (150 and 300 mM) and sizes of AgNPs and compared with those treated with standard glibenclamide. A significant decrease (P > 0.05) in the glucose level was achieved for 30, 45, and 65 nm after 15 days of treatment compared to the diabetic control. The oral administration of optimal AgNPs reduced the glucose level from 280.83 ± 4.17 to 151.17 ± 3.54 mg/dL, while the standard drug glibenclamide showed the reduction in glucose from 265.5 ± 1.43 to 192 ± 3.4 mg/dL. Histopathological studies were performed in dissected kidney and liver tissues of the treated mice, which revealed significant recovery in the liver and kidney after AgNP treatment. Acute toxicity study revealed that AgNPs were safe up to a size of 400 nm and the raw leaf extract of Emblica phyllanthus was safe up to 2500 mg/kg b.w. This study may help provide more effective and safe treatment options for diabetes compared to traditionally prescribed antidiabetic drugs.

Genetic association study of ERBB4 SNP rs1351592 with polycystic ovary syndrome in Pakistani population.

Polycystic ovary syndrome (PCOS) is an oligogenic condition characterised by hyperandrogenism, ovulatory dysfunction and polycystic ovarian morphology. Previously, European and Han Chinese populations identified different susceptibility loci, of which ERBB4 (rs1351592) was strongly associated with PCOS. Our study aimed to investigate the association of ERBB4 Single Nucleotide Polymorphism (SNP), rs1351592 with PCOS in Pakistani women of Hazara region. Fifty PCOS patients and 14 healthy women were recruited and SNP was replicated using ARMS-PCR and sequencing. The study showed that Luteinising Hormone (LH), Follicle Stimulating Hormone (FSH), and Testosterone (T) were significantly elevated in patients compared to controls (P <0.05). Overall, the frequency of G allele was higher than C allele and the SNP lacked significant association with PCOS. This is the first study demonstrating the association of ERBB4 SNP, rs1351592 with PCOS in Pakistani population. Further research using larger population size will help to estimate the role of ERBB4 SNP as potential biomarker for disease diagnosis.

Patterns of maternal nutritional status based on mid upper arm circumference.

OBJECTIVE: To determine the patterns of maternal nutrition status by using mid upper arm circumference (MUAC) and to examine the association of maternal nutritional status with the nutritional status of malnourished children under two years of age. METHODS: Descriptive study conducted at the Department of Pediatric Medicine of the Children's Hospital, Lahore from January 2017 to March 2018. A total of 227 mother accompanying their children admitted for nutritional rehabilitation were included. Demographics of participants along with MUAC of every mother was taken. Data analysis was done by SPSS 22. RESULTS: Mean maternal age was 28.29±5.30 years and mean age of children was 9.22 ± 6.05 months. Mean maternal MUAC was 25.53±3.63 cm. Normal nutrition was present in only 70 (31%), 35 (15.4%) had moderate to severe under nutrition and 68 (30%) were overweight and 17 (7.5%) were obese. Maternal illiteracy was common 150 (66%) and 203 (89%) belong to poor social class. Majority 150 (75%) children had <-3SD WHZ score. Only 42 (18.5%) children were exclusively breast fed. Maternal malnutrition was significantly associated with severity of child's undernutrition (p=0.045) and low rates of exclusive breast feeding practices (p=0.049). CONCLUSION: Malnutrition, in the form of both under nutrition and obesity is prevalent in mothers of malnourished children belonging to lower social class. Maternal illiteracy and low income are the major contributor in maternal malnutrition which in turn has an impact on child nutrition and breast feeding practices.

New Anti-inflammatory Triterpene Esters and Glycosides from Alstonia scholaris.

BACKGROUND: Phytochemical studies on the ethanolic extract of aerial parts of Alstonia scholaris lead to the isolation of two new triterpenoid of the lanostanetype, lanosta 5ene,24-ethyl-3-O-ß-D-glucopyranoside (1), lanosta,5ene,24-ethyl-3-O-ß-D-glucopyranosideester (2) and new ursane type triterpenoidmethylester, 12-ursene-2,3,18,19-tetrol,28 acetate (nighascholarene) (3), together with seven known triterpenes, betuline, triterpene of the lupane type, alstoprenyol (4), 3ß-hydroxy-28-ß-acetoxy-5-olea triterpene (5),α-amyrin acetate (6), α-amyrin (7), lupeol acetate (8), 3ß-hydroxy-24-nor-urs-4,12,28-triene triterpene (9) and ursolic acid (l0). METHODOLOGY: The triterpenoid structures of these colorless compounds were deduced from the 1H and 13C-NMR data, and in particular from the application of two-dimensional 1H, 13C correlation experiments as well as by comparison with reported literature data. CONCLUSION: This study deals with isolation and structural elucidation of natural new triterpenoidesters and glycosides with anti-inflammatory activity.

Potential effect of the microbial fermented feed utilization on physicochemical traits, antioxidant enzyme and trace mineral analysis in rabbit meat.

The study investigated the potential effect of the microbial fermented feed utilization on physicochemical traits, antioxidant enzyme and trace mineral analysis in rabbit meat. A total of 72 six-week-old male rabbits were weighed and randomly divided into four groups (1) (SRKC) control; (2) (SRKP) Lactobacillus plantarum 1 × 106  cfu/g fresh weight (FW); (3) (SRKG) Pediococcus acidilactici 1 × 106 cfu/g FW and (4) (SRKPG) P. acidilactici + L. plantarum 1 × 106 cfu/g FW. Performance characteristic, weekly body weight, was positively (p < .05) enhanced, while daily feed intake (DFI) and feed convention ratio (FCR) were not influenced in treatments group as compared to untreated. The water, protein, water holding capacity (WHC) and dry matter (DM) concentration were positively (p < .05) influenced, while ash, pH, lightness, redness and yellowness were not influenced in treated group as compared to untreated. The concentration of glutathione peroxidase (Gpx), superoxide dismutase (SOD) and aspartate aminotransferase (AST) was positively (p < .05) influenced in treatments group as compared to control. Regarding trace minerals, copper (Cu), iron (Fe), manganese (Mn) and zinc (Zn) were positively (p > .05) reduced in treated group as compared to untreated. It is concluded that the addition of lactic acid bacteria (L. plantarum and P. acidilactici) in Hybrid pennisetum silage had a constructive influence on rabbit health performance and meat biochemistry.

Prognostic factors and outcome of Post-Neonatal Tetanus in an intensive care unit of a Tertiary Care Hospital.

OBJECTIVE: To determine the prognostic factors and outcome of tetanus in children of post-neonatal age admitted in the intensive care unit (ICU) of a tertiary care hospital. METHODS: This prospective cross sectional study, carried out in the Pediatric ICU of The Children's Hospital Lahore from Jan 2013 to March 2017. Children of both genders with age range of two months to 16 years diagnosed clinically as tetanus were included. All 132 patients were scrutinized for all possible risk factors, need for mechanical ventilation and outcome. Data was analyzed by SPSS version 20. RESULTS: Mean age of children was 7.5±3.4 years with male predominance (70.5%). Only (38.6%) received three doses of vaccination but none had booster dose. Trauma (43.2%) encompassed maximum predisposing factor followed by ear or nose prick and ear discharge. Mean duration of ICU stay was 20±13.3 days. Mortality rate was (17.4%). Ventilator support was given to (78.8%). Neurological outcome was normal in (82.6%). Trauma, ear or nose prick in girls and ear discharge were significantly associated with poor outcome and death with p-value of <0.001, 0.011 and <0.001 respectively. Other factors associated with poor outcome were need for mechanical ventilation and neurological impairment with p-value of 0.001 and <0.001 respectively. CONCLUSION: Tetanus is causing our children to suffer from devastating disease. Vaccination status is not satisfactory and along with trauma, ear discharge and ear or nose prick are identifiable risk factors. To combat these issues large scale vaccination and booster doses remains promising option.

Biological and biochemical characteristics of male reproductive system, serum metabolites and carcass quality of Japanese quails by the supplementation of Pinus ponderosa leaves and α-tocopherol acetate.

The aim of the current study was to evaluate the effect of dietary supplementation of Pinus ponderosa leaves (pine leaves) and α-tocopherol acetate (vitamin E) powder on male reproductive system, serum metabolites and carcass characteristics of Japanese quails. A total of 360-day-old male quails were purchased from the open market and kept at poultry shed for ninety-four days. After ten days of adaptation, all quails were randomly assigned into 4 groups, control (IC); supplemented with α-tocopherol acetate (IE) at the rate of 150 mg/L; Pinus ponderosa leaves (IP) at the rate of 150 mg/L; and 70 mg α-tocopherol acetate and 70 mg Pinus ponderosa leaves (IEP). Pinus ponderosa leaves and α-tocopherol acetate supplementation had not significantly (p > .05) effected on final body weight gain, feed intake and feed conversion ratio of quails. The high-density lipoprotein cholesterol (HDLC) and total cholesterol (TC) were significantly (p > .05) affected by IE and IP groups as compared to IC and IEP groups. Triglyceride (TG), glutathione peroxidase (GPx) and superoxide dismutase (SOD) significantly (p < .05) increased in all treatment groups except for the IC group. Aspartate transaminase (AST) significantly (p > .05) decreased in treatment groups as compared to control group. Overall, the mineral levels significantly (p < .05) increased in treatment groups as compared to control. Cloacal gland index values, the quantity of foam production and testis weight were significantly (p < .05) increased in treatment groups. It was concluded that the supplementation of Pinus ponderosa leaves and α-tocopherol acetate improved the testis weight, foam production, serum antioxidant enzymes and mineral level especially zinc in Japanese quail considered an indicative characteristic of higher sperm production rate and improved sexual activity. Further, higher gametogenesis rate, sperm production or reproductive behaviour including different hormonal level will be analysed in future study.

Report: Occurrence of Aflatoxin M1 in raw and processed milk consumed in Pakistan.

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.

Antibiotic susceptibility profile of bacterial isolates from post-surgical wounds of patients in tertiary care hospitals of Peshawar, Pakistan.

Postoperative wound infections are the infections of the operating site within thirty days after surgery. The infections that develop after surgery are a major problem throughout the world leading to, increased morbidity and mortality. This study was carried out to determine the prevalence of bacterial pathogens causing wound infection in the surgical wards and to determine the antimicrobial sensitivity patterns of the isolated bacteria. A total of 250 wound samples were collected over a period of 6 months from July-December, 2016. The pathogenic bacteria were isolated, identified and their antibiotic susceptibility was determined through disc diffusion method. Among 250 cases, 210 (84%) were culture positive for bacterial pathogens, while 40 (16%) were bacteriologically sterile (Negative). Rate of infection was high in males (55.6%) than females (44.4%). The predominant isolates were E.coli 55 (26.19%), followed by S.aureus 51 (24.28%), Pseudomonas spp. 43(20.47%), S.aureus MRSA 21 (10%), Proteus Marbillis 15 (7.14%), E.coli ESBL producer was 8 (3.81%), Acinetobacter 7 (3.33%) Proteus valgaris 5 (2.38%), b-Streptococci 3 (1.43%) and Klebsella pneumonia were the least, 2 (0.95%). Linezolid, Vancomycin, Amoxycillin, Cefoperazone and Meropenem are the most effective antibiotics for treating post-surgical wound infections.

Age-related variations in the in vitro bactericidal activity of human sera against Pseudomonas aeruginosa.

The human serum is a vital component of the innate immunity of the host that acts as the first line of defence against invading pathogens. A key player in serum-mediated innate immune defence is a system of more than 35 proteins, collectively named as the complement system. After exposure of the pathogen, these proteins are activated in a cascade manner, ultimately forming a membrane attack complex (MAC) on the surface of the pathogen that directly lyses the bacterial cell. Formation of the MAC can be demonstrated in vitro by using serum bactericidal assay (SBA) that works in the absence of cellular components of blood after incubating the serum along with bacteria. Here, we describe the age-related differences in the bactericidal activity of human serum against Pseudomonas aeruginosa, an opportunistic human pathogen causing an array of hospital and community-acquired infections. We demonstrate that adult sera were highly effective in the in vitro killing of Pseudomonas aeruginosa as compared to children and the elderly (p < 0.0001). Sera from children were seriously compromised in the killing P. aeruginosa, whereas elderly sera showed a reduced level of killing. Data revealed a positive correlation between age and serum-killing with higher coefficient of determination values of 0.34, 0.27, and 0.58 and p values of < 0.0001, < 0.001, and < 0.0001, respectively, after 60, 90, and 120 minutes of incubation. Hence, our study highlights the age-related difference in the bactericidal activity of human sera. We conclude that sera of children are totally compromised, whereas elderly sera are only partially compromised, in the killing of P. aeruginosa.

Microbial biotransformation of bioactive and clinically useful steroids and some salient features of steroids and biotransformation.

Steroids are perhaps one of the most widely used group of drugs in present day. Beside the established utilization as immunosuppressive, anti-inflammatory, anti-rheumatic, progestational, diuretic, sedative, anabolic and contraceptive agents, recent applications of steroid compounds include the treatment of some forms of cancer, osteoporosis, HIV infections and treatment of declared AIDS. Steroids isolated are often available in minute amounts. So biotransformation of natural products provides a powerful means in solving supply problems in clinical trials and marketing of the drug for obtaining natural products in bulk amounts. If the structure is complex, it is often an impossible task to isolate enough of the natural products for clinical trials. The microbial biotransformation of steroids yielded several novel metabolites, exhibiting different activities. The metabolites produced from pregnenolone acetate by Cunning hamella elegans and Rhizopus stolonifer were screened against tyrosinase and cholinesterase showed significant inhibitory activities than the parent compound. Diosgenin and its transformed sarsasapogenin were screened for their acetyl cholinesterase and butyryl cholinesterase inhibitory activities. Sarsasapogenin was screened for phytotoxicity, and was found to be more active than the parent compound. Diosgenin, prednisone and their derivatives were screened for their anti-leishmanial activity. All derivatives were found to be more active than the parent compound. The biotransformation of steroids have been reviewed to a little extent. This review focuses on the biotransformation and functions of selected steroids, the classification, advantages and agents of enzymatic biotransformation and examines the potential role of new enzymatically transformed steroids and their derivatives in the chemoprevention and treatment of other diseases. tyrosinase and cholinesterase inhibitory activities, severe asthma, rheumatic disorders, renal disorders and diseases of inflammatory bowel, skin, gastrointestinal tract.

Dietary Exposure and Natural Occurrence of Total Aflatoxins in Basmati Rice of Pakistan.

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.