Trends and current food safety regulations and policies for functional foods and beverages containing botanicals.

Globally, the demand for functional foods and beverages has significantly increased due to socioeconomic changes, particularly in health consciousness to enhance their functionality. Functional ingredients derived from botanicals are widely used because of their phytochemical properties with health benefits. This study aims to (1) review the capabilities and challenges of botanical addition in functional foods, (2) review current policies and regulations for functional foods containing botanicals in the European Union (EU), Canada, Japan, the Republic of Korea, and Thailand, and (3) provide recommendations on effective food safety control measures for better consumer trust and trade facilitation. This critical review was analyzed from online publications and available guidelines, regulations, and control measures published by food industries and governments in the EU and the four selected countries. The result confirmed that potentialities of botanicals arise from numerous bioactive compounds with varieties of sources. However, the usage may potentially raise health risks through hazardous substances in different species or plant parts, contaminants from environments and uncontrolled processes. Inadequate knowledge of botanical formulation and the maximum limit for daily consumption may elevate health risks through food-drug interaction or adverse effect incidents. Current policies and regulations show that varieties of measures are implemented influencing both economic growth and consumer awareness. The novel finding is that countries that provide a comprehensive national food control system influence not only the growth of the functional food subsectors but also build trust in food safety among trade partners and consumers.

Antimicrobial Activities and Biopreservation Potential of Lactic Acid Bacteria (LAB) from Raw Buffalo (Bubalus bubalis) Milk.

The aim of this study was to investigate the antimicrobial and biopreservation potential of lactic acid bacteria. The potential probiotic culture inhibited the growth of gram-positive and gram-negative foodborne pathogens in agar spot assay with inhibition zones ranging from 10 to 21 mm in diameter. The strains showed coaggregation capabilities ranging from 7 to 71% with tested food pathogens including Escherichia coli, Staphylococcus aureus, Listeria monocytogenes, and Salmonella enterica subsp. enterica serovar Typhimurium. The effect of cell-free supernatants on the release of 260 nm absorbing material, especially nucleic acids, was evaluated and indicated the antagonistic activity on foodborne pathogens, the highest being Lactobacillus paraplantarum against E. coli (3.77) and S. aureus (3.86) after 60 min. The effect of cell-free supernatant (CFS) on the growth of pathogens showed that Lactobacillus paraplantarum 11 and L. pentosus 93 had the highest inhibitory activity against tested strains. The biopreservation assay indicated that the potential probiotic strains Lactobacillus paraplantarum 11 (BT), Lactiplantibacillus plantarum 19, Lactobacillus pentosus 42, Limosilactobacillus fermentum 60, Lactobacillus pentosus 93, and Limosilactobacillus reuteri 112 were effective in reducing the Listeria monocytogenes population in raw buffalo milk. Complete Listeria monocytogenes inhibition was observed after 6-8 days. This study showed that probiotic LAB from buffalo milk have antimicrobial and biopreservation potential; these strains have the potential to be utilized as biopreservative agents in food products.

Formulation of nutritional and ready to mix gluten-free porridge from proso millet (Panicum miliaceum) little millet (Panicum sumatrense) and legume faba bean (Vicia faba).

The underutilized millets and legumes (proso millet, little millet, soaked and germinated faba beans) were mixed in different combinations to develop ready to eat porridge. Physiochemical and functional properties such as color, water absorption index (WAI), water solubility index (WSI), pasting properties, microstructure and in vitro starch digestibility of formulated porridge mix were evaluated. The porridge mix formulations exhibited a considerable amount of proteins (17.97-22.17 g/100 g) and carbohydrates (70.24-72.25 g/100 g). WAI and WSI of the different porridge formulations (P1-P6) ranged from 3.07 to 3.34 g/g and 10.37 to 25.81 g/100 g/100 g, respectively. The in vitro starch digestibility of formulation P5 (little millet, proso millet and soaked faba bean) was significantly higher (67.70%) compared to other formulations. This study concluded that nutritional, functional values as well as the digestibility of porridge mixes were improved after incorporation of soaked faba beans. The ready to eat porridge mix based on underutilized legumes and millets can serve as source of macro and micronutrients for all age groups.

Comparative study of physicochemical and functional properties of soaked, germinated and pressure cooked Faba bean.

Faba bean flours (germinated, soaked, cooked and raw) were evaluated for physiochemical and functional properties. The flour samples exhibited considerable amounts of carbohydrates (58.79-66.19 g/100 g) and proteins (21.9-29.1 g/100 g). Soaked faba bean (SFB) (29.1 g/100 g) and raw faba bean (RFB) (25.70 g/100 g) flour contained significantly higher amount of protein than germinated faba bean (GFB) and pressure cooked faba bean (PCFB). The physicochemical and functional composition of GFB and PCFB were improved compare to raw flour. Physical and functional properties such as water absorption index (2.97 g/g) and foaming stability (140.13 mL/100 mL) were increased by germination. The functional properties of pressure-cooked faba bean such as water solubility index (2.12 g/100 g) and water absorption capacity (2.02 g/g) were higher than other flour samples. The microstructure of legume flour samples explained that the starch granules of raw flours were smooth, oval and granular structure whereas soaked, germinated and cooked flours showed damaged starch granules. The effect of soaking, germination and pressure-cooking demonstrated significant variations in functionals characteristics of faba bean flour. Therefore, various processing conditions can be combined to obtain the desired characteristics in faba bean-based food products.

Enhancing bioaccessibility and bioavailability of carotenoids using emulsion-based delivery systems.

The consumption of foods rich in antioxidants, vitamins, minerals including carotenoids etc. can boost the immune system to help fight off various infections including SARS- CoV 2 and other viruses. Carotenoids have been gaining attention particularly in food and pharmaceutical industries owing to their diverse functions including their role as pro-vitamin A activity, potent antioxidant properties, and quenching of reactive oxygen (ROS), such as singlet oxygen and lipid peroxides within the lipid bilayer of the cell membrane. Nevertheless, carotenoids being lipophilic, have poor solubility in aqueous medium and are also chemically instable. They are susceptible to degrade under stimuli environmental conditions during food processing, storage and gastrointestinal passage. They also exhibit poor oral bioavailability, thus, their applications in aqueous-based foods are limited. As a consequent, suitable delivery systems including colloids-based are needed to enhance the solubility, stability and bioavailability of carotenoids. This review presents challenges of incorporation and delivery of carotenoids focusing on stability and factors affecting bioavailability. Furthermore, designed factors impacting bioaccessibility and bioavailability of carotenoids using emulsion-based delivery systems are explicitly explained. Each delivery system exhibits its own advantages and disadvantages; thus, the delivery systems should be designed based on their targets and their further applications.

Protection and Controlled Gastrointestinal Release of Bromelain by Encapsulating in Pectin-Resistant Starch Based Hydrogel Beads.

Hybrid pectin and resistant starch-based hydrogel beads loaded with bromelain using the extrusion gelation method were prepared and evaluated to enhance the activity of bromelain during gastrointestinal passage and thermal processing. The solutions of pectin-resistant starch with bromelain were dropped into the gelation bath containing calcium chloride (0.2 M) solution to develop various types of hydrogel beads. The physicochemical characteristics of the synthesized hydrogel beads were evaluated. The ratio (4.5:1.5 w/w) of pectin and resistant starch concentration significantly (p < 0.05) enhanced the encapsulation efficiency (80.53%). The presence of resistant starch resulted in increased entrapment of bromelain, improved swelling properties with sustained release behavior, and improved gastric stability than pectin hydrogels alone. The swelling of hydrogel beads was higher at pH 7.4 than pH 1.2. Optimized batch of hybrid pectin/resistant starch exhibited a spherical shape. Optical and scanning electron microscopy showed a more packed and spherical shape from the pectin/resistant starch hydrogel bead network. Fourier transformation infrared spectroscopy was also used to confirm the presence of bromelain in the hydrogel beads. The encapsulated bromelain in the pectin/hi-maize starch beads produced at a pectin/hi-maize ratio of 4.5:1.5 (percent w/w; formulation P4) obtained the highest relative bromelain activity in all heat treatments including at 95°C, whereas the highest activity of free bromelain was found only at 30°C. Bromelain encapsulated in hydrogels released at a faster rate at simulated intestinal fluid (SIF, pH 7.4) than at simulated gastrointestinal fluid (SGF, pH 1.2).

Bioutilization of Chicken Feather Waste by Newly Isolated Keratinolytic Bacteria and Conversion into Protein Hydrolysates with Improved Functionalities.

Chicken feathers are major poultry waste that is difficult to process in its native form due to highly resistant keratin protein in large amounts. In this study, a novel feather-degrading bacterium, Bacillus amyloliquefaciens KB1, was screened from a chicken farm bed (CFB) using morphological and biochemical tests followed by 16s rDNA analysis. Among observed isolates, bacterial isolate (KB1) showed the highest degree of feather degradation (74.78 ± 2.94%) and total soluble protein (205 ± 0.03 mg/g). The optimum fermentation conditions obtained were at 40 °C (temperature), pH 9, and 1% (w/v) feather concentration using response surface methodology in a Box-Behnken design. It produced 260 mg/g of soluble protein and bioactive peptides with 86.16% feather degradation. The amino acid profile showed an increase in the concentration of essential amino acids compared with the feather meal broth. The selection of a safe screening source for this new bacterium in CFB produced hydrolysates with enhanced bioactivity applicable for feed, and cosmetic applications, along with environmental bioremediation.

Factors Influencing the Intensity of Adoption of the Roundtable on Sustainable Palm Oil Practices by Smallholder Farmers in Thailand.

The Roundtable on Sustainable Palm Oil (RSPO) was formed in 2004 in response to adverse environmental impacts caused by the expansion of oil palm production. It created voluntary standards to accelerate adoption of improved practices along the supply chain to ensure environmental benefits. However, the rate of adoption among smallholders of the practices RSPO recommends remains low. This study assessed the factors influencing smallholder farmers' adoption decisions and the intensity of adoption using primary data obtained from 723 randomly selected oil palm producers in the southern region of Thailand, of which 333 RSPO were adopters and 390 were non-adopters. The results of the binary logit and two-limit Tobit regressions revealed that total farm labor, farm size, number of training sessions attended, access to information, extension services, support services, and perceived ease of adoption of the practices had positive effects on the intensity of adoption. However, the age of oil palm trees negatively influenced the intensity of adoption. The results suggest that strengthening capacity building activities and extension services would increase the adoption of the RSPO practices among small-scale farmers.

Vitamin D3 enhanced intestinal phosphate cotransporter genes in young and growing broilers.

The influence of dietary vitamin D3 (VD3) levels on growth, bone performance, and duodenal type IIb sodium-dependent phosphate cotransporter (NaPi-IIb) genes in broiler chicken were studied. One-day-old male Ross308 broilers (n = 432) were allocated into 6 treatment groups with each group consisting of 6 cage pens. Each treatment group received diet containing different amounts of VD3 (80, 200, 500, 1,250, 3,125, or 7,813 IU per kg of diet) from a day-old to 31 D of age. Dietary available phosphorus and calcium were kept the same across all treatments in each phase. At 14 D, influence of VD3 on BW gain was found in the birds that received VD3 of 3,125 IU/kg and 200 IU/kg (P < 0.05). Toe ash and tibia ash linearly increased (P < 0.05) at 14 D with increase in dietary VD3. There was no significant influence of dietary VD3 on tibia breaking strength. In both phases, relative expression of duodenal NaPi-IIb linearly increased (P < 0.01) with increase in dietary VD3. At 14 D, highest expression of 3.2 folds was observed in birds treated with VD3 at 7,813 IU/kg of feed. At 31 D, birds that received VD3 levels of 3,125 and 7,813 IU/kg of feed showed 2.9 folds higher in NaPi-IIb expression compared with those fed lowest level of VD3 at 80 IU/kg of feed. When dietary calcium and phosphorus were maintained at the standard requirement, increase in dietary VD3 did not improve growth performance. For optimum growth and bone characteristics, dietary inclusion of VD3 at 500 IU/kg was adequate for both starter and grower broiler diets. Vitamin D3 enhanced the expression of NaPi-IIb at higher doses and thus improving the tibia ash content in high VD3 treatment groups. This study reported for the first time an increased in the expression of duodenal NaPi-IIb in 31-day-old broilers in response to high dietary VD3 levels.

Enhanced yield of Scenedesmus obliquus biomacromolecules through medium optimization and development of microalgae based functional chocolate.

The freshwater green microalga Scenedesmus obliquus was cultivated to enhance the contents of proteins, carbohydrates and lipids by using Box-Behnken experimental design. S. obliquus was cultured under phototrophic conditions by using Bold's Basal Medium with different cultivation parameters including pH (7, 8 and 9), salinity (10, 30 and 50 mM), and nitrogen source (0.125, 0.5 and 1 g/L). The highest biomass yield (64.9 ± 0.94 mg/L/day) was obtained by using optimized medium at a salinity concentration of 30 mM (w/v), and nitrogen sources of 0.125 g/L. The maximum content of protein, lipid and carbohydrates from S. obliquus optimized medium were 342.19 ± 0.28 mg/g, 241.41 ± 4.32 mg/g and 288.05 ± 1.12 mg/g of dry wt. respectively. The amino acid and fatty acid analysis of S. obliquus biomass indicated the presence of significant amount of essential amino acids and essential fatty acids. Furthermore, chocolate crispy bar was developed by fortification with encapsulated freeze-dried S. obliquus and evaluated for its oxidative stability and sensory analysis. The chocolate fortified with microalgae can be a potential source of essential fatty acids and amino acids in addition to other bioactive compounds.

Effects of Lintnerization, Autoclaving, and Freeze-Thaw Treatments on Resistant Starch Formation and Functional Properties of Pathumthani 80 Rice Starch.

The objective of this study was to assess the effects of lintnerization, autoclaving, lintnerization followed by autoclaving, and freeze thawing treatments on the production of resistant starch from Pathumthani 80 (RD 31) rice. The produced resistant starch was further evaluated for some important physicochemical properties including pasting properties, swelling behavior, digestibility, water holding capacity, and functional properties including glycemic index and antioxidant properties. The lintnerization treatment and autoclaving significantly (p ˂ 0.05) increased resistant starch content to 64% (w/w) and gave the lowest glycemic index (46.12%). The lintnerization followed by autoclaving treatment significantly increased the solubility and water holding capacity, reduced the swelling power, and disrupted the crystalline structure of the starch granules. The native rice starch with autoclave treatment exhibited the highest swelling power among the samples, while the acid hydrolyzed starch was followed by autoclave treatment showing the lowest swelling power (1 g/g) at 90 °C. Fourier transform infrared analysis revealed the modified structures and bonding of the starch materials with the shifting of C=O stretch. However, the antioxidant properties and pasting properties were observed to decrease with the lintnerization, autoclaving, and freeze-thawing treatment of the native starch. The highly resistant starch content and low glycemic index value of the autoclaved RD 31 starch indicates the potential of the resistant starch's application for the formulation of functional foods targeting the diabetic population.

Restriction of dietary non-phytate phosphorus on growth performance and expression of intestinal phosphate cotransporter genes in broilers.

Effects of dietary non-phytate phosphorus (nPP) restriction on growth and duodenal type IIb sodium-dependent phosphate cotransporter (NaPi-IIb) genes were observed. A total of 432 one-day old Cobb500 male broiler chickens in 36 cage pens were divided into 6 groups with each group containing 6 pens. Each group was treated with one of the diets containing 0.33, 0.37, 0.41, 0.45, 0.49, and 0.53% of nPP up to 14 D. During 15 to 31 D, birds were treated with one of the diets containing 0.23, 0.27, 0.31, 0.35, 0.39, and 0.43% of nPP. Level of Ca was kept the same across all treatments. Dietary nPP level influenced (P < 0.001) weight gain and feed intake in both growth phases, whereas effect on feed per gain ratio was seen only in the second phase. Toe ash, tibia ash, and tibia breaking strength responded to treatments (P < 0.01) at 14 D. Only tibia ash content was significantly improved (P < 0.001) at 31 D. Growth and bone parameters linearly improved with an increase in dietary nPP content (P < 0.05). Above dietary nPP 0.41% and 0.31% for first phase and second phase, respectively, no significant improvement was seen. Duodenal NaPi-IIb mRNA overexpressed with a decrease in dietary nPP in both phases (P < 0.05). Relative expression of NaPi-IIb in lowest nPP group were 2.2 folds higher in the first phase and 3.6 folds higher in the second phase compared to respective highest nPP groups of each phase. No significant change in NaPi-IIb expression was seen above 0.37% of dietary nPP for 14 D and 0.31% of dietary nPP for 31 D. Dietary requirements of nPP 0.41% for 0 to 14 D and 0.31% for 15 to 31 D were adequate for optimal growth and bone parameters. This study fills the gap in understanding of intestinal NaPi-IIb expression in response to dietary nPP restriction in broilers older than 21 D of age.

Survival and Behavior of Encapsulated Probiotics (Lactobacillus plantarum) in Calcium-Alginate-Soy Protein Isolate-Based Hydrogel Beads in Different Processing Conditions (pH and Temperature) and in Pasteurized Mango Juice.

Hybrid alginate-soy protein isolate-based hydrogel beads were prepared and evaluated to enhance the survival of the encapsulated probiotics (Lactobacillus plantarum) during heat processing to incorporate in mango juice. The solutions of sodium alginate-soy protein isolate (SA-SPI) with probiotic cells were dropped into the gelation bath containing calcium chloride (3% w/v) solution to develop various types of hydrogel beads. The level of survival of probiotics in encapsulated beads under acidic conditions (pH 2, 3, and 6.5) and bile salt (0.5 and 1.0% w/v) was evaluated. The survival of the encapsulated probiotics to thermal processing was evaluated by treating the beads in saline solution (0.9% w/v) at 30, 50, 63, and 72°C. The encapsulated probiotic bacteria were found alive even after treatment at 72°C for 90 s. Most of the free cells did not survive at the temperature higher than 50°C and very low pH (pH 2 and 3). The survival of probiotic cells was found higher with the hybrid hydrogel beads containing alginate and soy protein isolate (1:8 w/w). Furthermore, mango juice fortified with encapsulated L. plantarum in hydrogel beads was subjected to thermal pasteurization at 72°C for 90 s.

Enhanced lipid and starch productivity of microalga (Chlorococcum sp. TISTR 8583) with nitrogen limitation following effective pretreatments for biofuel production.

The study was conducted to evaluate the conditions to enhance the accumulation of lipids and starch in Chlorococcum sp. TISTR 8583 for the production of biofuel. The Chlorococcum sp. TISTR 8583 was cultivated on BG-11 medium under optimized light intensity. The nitrogen limitation (NL) enhanced the accumulation of both starch and lipids and resulted in 34.02% total sugars as compared to 22.57% on nitrogen supplemented (NS) media only. Similarly, the nitrogen supplemented (NS) media produced 17.05% lipids as compared to 29.59% lipids by NL media. The biomass was investigated for biodiesel and bioethanol production by adopting different pretreatment strategies, such as enzyme, acid and alkaline pretreatments. The alkaline pretreatment was found to be efficient strategy (23.67 wt% sugars/g algal biomass: 1.2% (w/v) at 140 °C for 30 min) while the acid pretreatment (1%: v/v; 140 °C) was least effective pretreatment strategy with the yield of 14.83 wt% sugars/g algal biomass.

Culled banana resistant starch-soy protein isolate conjugate based emulsion enriched with astaxanthin to enhance its stability.

The conjugates of biomacromolecules such as proteins and polysaccharides have potential to stabilize the emulsion system and encapsulate valuable bioactive compounds for biofortification in food systems. In this study, native banana starch (NBS) was isolated from green culled banana and modified into resistant starch (type III) by lintnerization followed by autoclaving-cooling process, resulting in lintnerized-autoclaved banana starch (LABS). Soy protein isolate (SPI) was used for developing the polysaccharide-protein conjugates i.e. LABS-SPI conjugate and used as wall material to stabilize the oil-in-water emulsion system. LABS-SPI conjugate emulsions were subjected to in vitro digestion model and oxidative stability evaluation. Furthermore, the emulsion system was enriched with astaxanthin and evaluated for its stability. The chemical finger printing of LABS-SPI conjugates showed stretching in immine and enaminol group of Schiff's bases, the CN stretching of Amadori product. During in vitro digestion LABS-SPI conjugate emulsion showed that the presence of resistant starch had an influence on the droplet digestion process and significantly (p < 0.05) lower free fatty acid release compared to emulsions stabilized by SPI alone. LABS-SPI conjugate emulsion system demonstrated higher stability of astaxanthin at storage temperatures (6, 20 and 37 °C), and can be used for biofortification of food and pharmaceutical formulations.

Experimental investigation on biochemical, microbial and sensory properties of Nile tilapia (Oreochromis niloticus) treated with moringa (Moringa oleifera) leaves powder.

Six storage trials were conducted for preservation of Nile tilapia (Oreochromis niloticus) in ice storage at 5 °C using moringa leaves powder. The study aims to prolong the storage life of tilapia fish using moringa leaves powder of different concentrations. The effect of the moringa leaves powder (MLP) on biochemical, microbiological and sensory properties of fresh Nile tilapia stored in ice at 5 °C was investigated. Total volatile basic nitrogen, pH and thiobarbituric acid reactive substances values in MLP treated tilapia fish were 18.18 mg/100 g, 6.57, 5.63 mg MDA/kg respectively, which were significantly lower compared to untreated fish. Bacterial count (total viable count) was delayed significantly by increasing MLP concentration. The treated sample remains acceptable during storage in ice at 5 °C up to 11 days, whereas shelf life of untreated sample of tilapia was not extended beyond 7 days. The corresponding microbiological assessment also supports the results of sensory assessment that increased in shelf-life. The results revealed that the MLP is natural preservatives for extending the shelf life of Nile tilapia during transportation and ice storage.

Reinforced cassava starch based edible film incorporated with essential oil and sodium bentonite nanoclay as food packaging material.

Biodegradable packaging in food materials is a green technology based novel approach to replace the synthetic and conventional packaging systems. This study is aimed to formulate the biodegradable cassava starch based films incorporated with cinnamon essential oil and sodium bentonite clay nanoparticles. The films were characterized for their application as a packaging material for meatballs. The cassava starch films incorporated with sodium bentonite and cinnamon oil showed significant antibacterial potential against all test bacteria; Escherichia coli, Salmonella typhimurium and Staphylococcus aureus. Antibacterial effect of films increased significantly when the concentration of cinnamon oil was increased. The cassava starch film incorporated with 0.75% (w/w) sodium bentonite, 2% (w/w) glycerol and 2.5% (w/w) cinnamon oil was selected based on physical, mechanical and antibacterial potential to evaluate shelf life of meatballs. The meatballs stored at ambient temperature in cassava starch film incorporated with cinnamon oil and nanoclay, significantly inhibited the microbial growth till 96 h below the FDA limits (106 CFU/g) in foods compared to control films that exceeded above the limit within 48 h. Hence cassava starch based film incorporated with essential oils and clay nanoparticles can be an alternate approach as a packaging material for food industries to prolong the shelf life of products.

Extraction and characterization of proteins from banana (Musa Sapientum L) flower and evaluation of antimicrobial activities.

Ultrasonic assisted alkaline extraction of protein from banana flower was optimized using response surface methodology. The extracted proteins were characterized by Fourier transform infrared spectroscopy and molecular weight distribution was determined by gel electrophoresis. The maximum protein yield of 252.25 mg/g was obtained under optimized extraction conditions: temperature 50 °C, 30 min extraction time and 1 M NaOH concentration. The alkaline extraction produced a significantly high protein yield compared to enzymatic extraction of banana flower. Chemical finger printing of proteins showed the presence of tyrosine, tryptophan and amide bonds in extracted protein. Alkaline and pepsin assisted extracted banana flower proteins showed characteristic bands at 40 and 10 kDA, respectively. The extracted proteins showed antibacterial effects against both gram positive and gram negative bacteria. The high protein content and antimicrobial activity indicate the potential applications of banana flower in the food and feed industry.

Preparation of eggshell membrane protein hydrolysates and culled banana resistant starch-based emulsions and evaluation of their stability and behavior in simulated gastrointestinal fluids.

This study aims to evaluate the effect of resistant starch (RS) derived from culled banana on the stability and characteristics of emulsions stabilized by eggshell membrane (ESM) protein hydrolysates. It was observed that incorporation of resistant starch improved the properties of the emulsions. Furthermore, the most stable emulsions developed (using a combination of RS and pretreated alcalase hydrolysates) were assessed for their behavior in an in-vitro gastrointestinal model system and changes in their particle size, zeta potential and morphology were evaluated. It was seen that these emulsions underwent flocculation and coalescence in the presence of pepsin and at higher concentrations of mucin enzyme and further coalescence and fatty acid release were observed after their passage through the small intestine. These insightful results about emulsion behavior in the gastrointestinal tract can be useful for designing delivery systems for controlled release of bioactive compounds.

Screening of antibiotic residues in fresh milk of Kathmandu Valley, Nepal.

The prevalence of two groups of antibiotics; namely penicillin and sulfonamides was studied in fresh milk available in Kathmandu Valley of Nepal. The milk samples (n = 140) were collected from three different sources; individual farmers, cottage dairies and organized dairies of Kathmandu valley. Qualitative and semi-quantitative analysis with rapid screening kits revealed that 23% samples were positive for antibiotic residues in the fresh milk for penicillin and sulfonamide groups (1-256 µg/kg). High performance liquid chromatography (HPLC) analyses detected 81% samples positive for amoxicillin (68-802 µg/kg), 41% for sulfadimethoxine (31-69 µg/kg), 27% for penicillin G (13-353 µg/kg), and 12% for ampicillin (0.5-92 µg/kg). Due to the precision and accuracy of liquid chromatography method, it detected more positive samples and consequently presented higher prevalence than the rapid screening kits. The antibiotic residues were found above the maximum residue limits that presented serious threat to consumer health and raised a serious concern regarding the implementation and monitoring of international regulations in developing countries.