Journals on Biomedical Imaging

Impact of Die Configuration on Physio-Chemical Properties, Anti-Nutritional Compounds, and Sensory Evaluation of Multi-Based Extruded Puffs

Introduction

Millets are highly valued by vegetarians, vegans, and individuals with coeliac disease for their exceptional nutritional benefits [1]. These plant-based foods are abundant in proteins and complex carbohydrates while being low in fat [2]. Additionally, they are a rich source of dietary fiber, essential minerals, and B-group vitamins [3,4]. However, Millet preparation can be a time-consuming process as they require prolonged soaking and cooking, which reduces their consumption by consumers [5]. To overcome this challenge, novel food products like – ready-to-eat Millet-based snacks could be developed. Such products could encourage more individuals to incorporate Millet into their diet [6]. The food processing technique of extrusion- cooking is highly versatile, multifunctional, and cost-effective. Raw materials are exposed to heat, pressure and shear forces during this process, leading to various biochemical reactions such as protein denaturation, starch gelatinization, fiber degradation, amylose-lipid complex formation through Maillard reaction [7]. Extrusion-cooking is commonly employed to create expanded snacks that are ready-toeat, come in different shapes and textures, and boast enhanced flavour and colour [8]. High concentrations of raffinose-family oligosaccharides (RFOs) in lentils cause stomach discomfort and reduce lentil quality for human consumption.

To develop strategies for lentil quality improvement, variability, heritability and effects of environmental conditions on the content and composition of soluble carbohydrates in lentil seeds have been investigated in detail [9]. Over the past few years, there has been significant research on creating extruded products using Millets [10]. In most cases, these Millets are blended with wheat [11]. Numerous studies have examined the impact of various extrusion-cooking factors, including feed moisture, screw speed, extrusion temperature, nutritional characteristics [12] physico-chemical, sensory and textural properties of prepared product. These studies have also included trials conducted directly at the industrial level [13,14]. However, there has been a lack of research on how the die configuration, which determines the final product’s shape and size, affects the extrusion of Millets. Food shape and size play a critical role in capturing the consumer’s attention [15]. These features strongly influence the implicit associations with consumers concerning nutritional value [16]. Moreover, the shape and size of food can significantly impact the sensory qualities and physico-chemical properties of the extruded product [17]. Therefore, the present study was designed to examine the impact of star shaped and circular die configurations, which have two different diameters of 35.9 and19.6 mm², respectively, on the anti-nutritional component, physico-chemical properties, and sensory features of extruded snacks made from Millet flour.

Materials and Method

For the development of extruded puff products, germinated pearl Millet (Pennisetum glaucum), finger Millet (Eleusine coracana), sorghum (Sorghum bicolour), foxtail Millet (Setaria italic) and rice (Oryza sativa) as base were used as raw materials for the present study work. These Millets were procured from a nearby local market in Meerut, Uttar Pradesh [18].

Optimization of Single Screw Extruder Operating Parameters for Different Millet Based Extruded Puffs Products

The operating parameters of the twin screw extruder mainly, the temperature, feed moisture content and the screw speed, were optimized for the various “best selected” four Millet fortified with rice based extruded puff products formulation. Three feed moisture content (20%, 25%, and 30% wt.b.), screw speeds (180, 270, and 360 rpm) and 160, 180°C and 270°C temperature were selected to produce extrudes. The extruded samples were then analyzed for physical analysis like expansion ratio, water holding capacity, moisture content, puffing properties and texture analysis. Further experiment was carried out at three different level of temperature (100, 110 and 120°C) with keeping constant screw speed and feed moisture content.

Sample Preparation from Extrusion

Laboratory scale single-screw extruder (Model no: GTL-100), (Zigmo Agro Pvt. Ltd. New Delhi, India) was used for sample preparation. During each extruder run, the extruder machine was allowed to equilibrate for 5-10 min until a stable torque was achieved. Extruded samples were collected on metal screens to allow excess steam to flash off. The extruded samples were collected in a low-density polyethylene bag after cooling and stored in a cool and dried area.

Development of Extruded Puff Products Based on the Millet’s Combinations

Extruded puffs preparation was done, and it mainly consists of puffs mixture of Millets in combination as shown in Tables 1 & 2 and rice is used as base. The blended puffs mixes at appropriate moisture content were extruded to produce extruded puff through the extrusion machine (manufacture: Jas Enterprises, Ahmadabad, India) into desired shaped products and developing extruded puff products based on Millet combinations involves a series of steps to create a desirable texture, flavor, and nutritional profile the process flow chart shown in Figure 1.

Multi Millet Conditioning

To achieve the optimal moisture concentration for extrusion (16 g/100 g), the Puffs were conditioned. The amount of water required to reach this moisture level was determined based on the initial moisture content of each flour type, which was 12.95 ± 0.01, 10.87 ± 0.01, and 12.54 ± 0.06 for pearl Millet, finger Millet and foxtail Millet, respectively. Water was added gradually to the flour in dough mixer at average speed to avoid the lumps formation. The process took approximately 20 minutes to obtain uniformly hydrated flour.

Extrusion-Cooking Process

Extrusion cooking is a food processing technology that integrates multiple operations such as mixing, cooking, kneading, shearing, shaping, and forming [19]. The DSE30 Lab Twin-screw extruder was used to carry out the extrusion-cooking process with a 12 kg/h capacity. The extruder machine design had two 38CrMoAl screws, a 5-kW motor and operating at maximum screw speed of 500 rpm, with three heating zones at 55, 95, and 125°C, respectively. The extrusion process was carried out using circular and star-shaped dies holes with cross-sections of 19.6 mm2 and 35.9 mm2, respectively. The circular die nozzle and star cross-section had a length of 6.35 mm. The feed rate was set to 2.5 g/s, the die temperature to 160°C, and the screw speed to 230 rpm. To examine the extruded products, the water absorption index, water solubility index, starch gelatinization degree, colour, phytate and oligosaccharides content were measured. For this purpose, extrudates were ground using an electrical grinder (HM- 5735) from Home Electrical Appliance and passed through a 0.25 mm sieve sized blades for some analysis, while other analyses were performed on the entire extrudates.

Physical Assessment of Extruded Puff

Finger Millet (Eleusine coracana L.), pearl Millet (Pennisetum glaucum), foxtail Millet (Setaria italic) and sorghum (Sorghum bicolour) are three selected cereals that have been evaluated in terms of physical parameters, specifically bulk density, in the context of cereals and cereal-based products. The evaluation of bulk density for the three combinations of puff products made from the selected cereals would involve measuring the mass and volume of the cereal products. The mass can be measured using a scale, while the volume can be determined by various methods such as displacement or geometric calculations. By studying and analysing the obtained results of bulk density, these conclusions can help in understanding the texture, density, and overall quality of the products, which are important factors for consumer acceptance. It’s worth noting that in addition to physical parameters like bulk density, other nutritional and sensory attributes of these cereals and their products are also crucial for evaluating their overall nutritional value and consumer appeal [20]. These may include factors such as nutrient composition, digestibility, taste, aroma, and shelf life, among others. By conducting such evaluations, can gain insights into the potential of these cereals and their products as staple foods and nutrient sources, both in developed and developing countries. Kjeldahl digestion methods (Advanced and ordinary) compared total nitrogen in non-germinated and germinated extruded puff products. Advanced Kjeldahl showed protein variations of (07.11% to 10.66%) and (09.45% to 11.98%), while ordinary Kjeldahl showed (05.11% to 10.96%) and (07.45% to 11.56%). Millet combinations revealed T3 with germinated Millets had higher protein, indicating nutritional value. Total ash, dietary fiber, and carbohydrates varied in extruded puff products. Statistical analysis indicated significant differences (p<0.0001) for each combination. Incorporating Millet and sorghum in rice flour (T3) resulted in higher protein and overall acceptable sensory properties. AOAC methods revealed significant differences (p=0.05) in proximate compositions of multi-Millet puffs with overall acceptable sensory properties [20]. This knowledge can be valuable for food scientists, policymakers, and nutritionists in promoting sustainable and nutritious food options worldwide.

Bulk Density and Expansion Ratio

Extruded products bulk density (BD) was studied by using the rapeseed displacement method and calibrated as per given equation (1) by [21]. For this purpose, the (10 ± 0.1 g) of randomly selected extruded product were weighted:

Whereas Wt. and Vep-1 are the weight in (gm) and the equivalent volume (cm3) of the extruded products, respectively. Actually, Vep is multiplication ratio between the rapeseed density (ρs) and the rapeseed weight (Ws), with same volume as the extrudates. Five replicates for each sample were prepared to study the parameters. sectional expansion index (SEI), bulk density (BD), instrumental color, and dry and bowl-life texture were evaluated. SEI and BD responses were fixed by moisture variable is incorporated; darker products were produced. Nevertheless, high together with higher temperature and lower moisture levels produces better color appearance. At high ratio desirable low hardness and crispy expanded extrudates can be generated as long as moisture is lower than about 22% [22]. The calibration of expansion ratio (ER) was according to the ratio of extruded product diameter to determine the die hole diameter of extruder with the help of caliper device, as reported by Kokseland Masatcioglu. For this purpose, ten replicates were taken for (ER) assessment.

Water Absorption Activity

The extruded products water absorption index (WAI) and the water solubility index (WSI) were determined as per equations (2) and (3) [23].

The (WSI) is the dry solid weight in the extracted supernatant, whereas (WAI) is the sediment weight without the supernatant per unit weight of the sample analyzed. Each sample was tested in a triplicate manner.

Starch Gelatinization

The starch gelatinization (SG) of the extruded products determination follows the modified method based on the procedure outlined by [24]. This method involves the formation of a blue iodine complex when amylase is released during gelatinization. Specifically, 40 mg quantity of sample was dissolved in a 50 mL of 0.15 M KOH solution and mixed thoroughly for 15 minutes. The reaction mixture was then centrifuged at 4032 x g for 10 minutes to remove out insoluble sediment. Next, 1 mL quantity of the supernatant was neutralized by the addition of 9 mL of 0.017 M HCl, and 0.1 mL of iodine reagent (1 g iodine and 4 g potassium iodine dissolved in 100 mL water). The resultant solution was mixed, and read the absorbance at 600 nm (A1) using a Cary 60 UV-VIS spectrophotometer. In addition, 1 M KOH and 0.1 M HCl solution was used to prepare control samples. The overall value of DG was calibrated by using equation (4) based on the average of three replicates.

Whereas (A1/A2) is the ratio of absorbance at 600nm of the sample to that of the control.

Phyto-Chemical Constituents of Selected Millets Puffs

The effect of extrusion processing on anti-nutrients factors namely tannin, phytate and saponin of extruded Millet-sorghum blend rice puffs puff product combinations were studied. Extrusion processing is necessary to absorb essential micro and macronutrients from added blends of puffs during extruded snacks preparations and eliminates a negative effect of anti-nutritional factors such as tannin, phytate and saponins. Thus, nutritional quality was maintained by extrusion through destruction of anti-nutritional components.

Color Determination

The CM-600d colorimeter (Konica Minolta Sensing Inc., Osaka, Japan) was used to determine the lightness (L*), redness (a*), and yellowness (b*) of both Puffs and extruded products with help of using the Spectra-Magic NX software (Konica Minolta, Tokyo, Japan). The series of experiments was replicated five times in a row.

Oligosaccharides Analysis

The presence of oligosaccharides namely verbascose, stachyose, and raffinose in Puffs and extruded products were investigated by using the HPLC with slight modifications in a method prescribed by [25]. Samples were mixed with deionized water, filtered, and separated isostatically on a cation exchange column. Identification was based on standard comparison, and quantification was based on calibration curves. Results are expressed in (mg/g dry matter) of each oligosaccharide after triplicate analysis.

Phytate Analysis

The phytate content of both Puffs and extruded products was determined by following the [26] method as its values were expressed in (mg/g) of dry matter of phytic acid. To calculate the phytate content, obtained values were multiplied by 0.282 and it is the molar ratio of phytate-phosphorus in a molecule of phytate. The analysis was done in a triplicate manner.

Sensory Evaluation

A panel of 28 semi-trained judges from University of Life Sciences and Technologies demonstrate Millet-based extruded snacks using a ranking test. Each sample is arranged in groups of three on glass plates to evaluate appearance, texture, taste, and aftertaste using an evaluation form as this form is generated through Fizz Acquisition 2.51 software. Warm black tea was used for taste neutralization between samples. The panelists ranked the samples as per most liked=1 to least liked=8 and the recorded results were calibrated as the summation of ranks for each sample (Figure 2).

Statistical Analysis

The recorded data of Millet Puffs and extruded products undergo one-way ANOVA and two-way ANOVA stats analysis, followed by Tukey’s HSD test. The two-way ANOVA stats analysis considers two major factors namely Millet puffs type and die type for further analysis. Minitab-17 ver. statistical software (Minitab, Inc., State College, PA, USA, 2010) was used to determine the significant differences among all studied parameter values at p < 0.05. The sensory evaluation data were statistically analysed by using the Friedman test with Fizz calculation (Biosystems, Cousteron, France) and a p<0.05 level of significance.

Results and Discussion

The various multi-Millet utilized to produce extruded snacks were shown in Table 1. Table 1 showed the notable differences among (L*, a*, and b*) colour patterns. Pearl Millet flour exhibited the highest a* and b* values while it had lowest L* value. In contrast, foxtail Millet flour had lightest colour, followed by finger Millet flour. Foxtail Millet flour exhibited the lowest a* and b* values, with the latter being statistically insignificant in comparison to pearl Millet flour. Additionally, Table 2 calibrated values were significantly differed (p<0.05) as the amount of anti-nutritional compounds were studied among the Puffs. Millets are known to contain several anti-nutritional compounds, including non-digestible oligosaccharides and phytic acid, which has inherent chelating characteristics to capture important divalent cations such as Fe, Zn, Ca, and Mg, which leads to lowering of availability for absorption and use in the small intestine [27,28]. However, presence of raffinose, verbascose, and stachyose oligosaccharides in human diets in an ample amount causes flatulence and discomfort in humans after consumption [29,30].

Note: *Each (n = 3) replica values were expressed as (mean ±standard deviation).

Note: *Each parameters value was expressed as (mean ± standard deviation) for (n=3).

Pearl Millet flour exhibited the highest concentration of phytates, followed by finger and foxtail Millets Puffs, respectively. Foxtail Millet flour contained the highest levels of verbascose, although it had stachyose in a lesser amount. Similarly, finger Millet flour had the highest amount of stachyose, while it has lowest amount of raffinose. However, the quantity of oligosaccharides in Millets varied as per the selected species and varieties, as well as the existing environmental conditions [31,32]. [33] reported significant variability in the amount of raffinose (4.10–10.30 mg/g), stachyose (10.70–26.7 mg/g) and verbascose (0.00–26.70 mg/g) among 18 different pea varieties. While Tahir et al. (2011) observed higher stachyose levels than raffinose and verbascose in 11 lentil varieties, as these findings were found in line with current findings. This difference in oligosaccharides level might be due to significant constraint on the extensive use of legumes at both domestic and industrial scale [34] (Figure 3).

Physico-Chemical Properties of Extruded Products

All physico-chemical parameters had significant differences (p < 0.05) among the extruded products, except water absorption index (WAI). These noticeable differences were attributed due to the type of Millet used, the type of die, and the (Millet x die) interaction, as shown in Table 3. However, it should be noted that the type of die had no significant effect on the WAI. Pearl Millet-based extruded products had the highest bulk density (BD) and water absorption index (WAI), whereas it has lowest expansion ratio (ER) and water solubility index (WSI) values. Similarly in foxtail Millet-based products had the highest (ER) and well-expanded spherical extrudates, while highest degree of starch gelatinization (SG) was recorded in finger Millet-based products. Higher ER, DG and WSI with lower BD was recorded in circular die configuration based extruded products as compared to star-shaped die. The effect of die shape on extruded product characteristics can be explained by change in friction and pressure strength. The circular die had a smaller cross-section, resulting in higher levels of friction and pressure and thus higher temperatures, which led to more expansion and lesser dense products, will obtain as it has greater ER and lower BD. On the other hand, the star-shaped cross-section had angles that could have mechanically broken bubbles in the gelatinized starchy matrix, which led to disturbance in expansion. However, with increment in the die nozzle diameter decreased radial expansion in yellow corn extrudates. Along with this, inducing higher extrusion pressure on starch gelatinization causes expansion of the extrudates products made from foxtail and finger except in pearl Millet-based products since it has higher fibre content that restricted starch gelatinization [1], [35]. Although ER and BD are important physical parameters that can influence consumer behavior in the sense of acceptability of extruded products [36].

Note: *Each parameters value was expressed as (mean±standard deviation) for (n=3).

Earlier studies by [37] who reported an inverse relation between BD and ER, as observed in this work, where a negative correlation was found (r = -0.631; p = 0.093). The presence of huge number of fibres in the feed Puffs can also affect the ER and BD values by reducing overall expansion on account of cell-wall rupture, resulting in a compact and hard product with an undesirable texture. In this context, Red lentil flour, with the highest fibre content, led to extrudates with the lowest ER and highest BD indicating the importance of considering fibre content while developing extruded product as described by [38]. The extrusion-cooking conditions, including the die used, may also impact the WAI and WSI values, which represent the amount of water that can be absorbed by the extruded product and the quantity of soluble substances formed during the extrusion process from starch, proteins, and fibres. The presence of fibres in higher amount could also influence functional properties [39]. Instead of this, WSI was also influenced by other factors such as legume type, die shape, and legume-die interaction, while effect of die shape had no significant effect on WAI. Higher fibre levels in the legume led to an increase in WAI, as they absorbed and retained water within a well-developed starch-protein-polysaccharide network, as discussed previously by [40].

Extruded Products Color

Colour is a crucial aspect of food products that can significantly affect consumer acceptability. Extrusion-cooking affects the colour features of the products, making them darker than the Puffs. The colour components of the extrudates were influenced by Millet type, die configuration and their interaction. The observed colour features were attributed to existing pigments and the Maillard reaction occurring during extrusion-cooking (Table 4). Star-shaped extrudates had greater L* and lower a* values (except for pearl Millet) compared to spherical shape extrudates, while b* index had found non-uniform trend. This fluctuating trend is due to pigment degradation as temperature rises and shear stress during extrusion is responsible to alter color, especially for carotenoids. The decrement in L* and increment in a* values may be linked to the melanoidins formation during the Maillard reaction, while increment in b* may be results from the formation of yellowish compounds during the initial stages of the Maillard reaction or from lipid oxidation. An advantage of larger die cross-section reduces extrusion pressure and heat, leading to a less intense Maillard reaction and reduces browning and flavor development [41]. Thus, star-shaped extrudates were obtained with the help of larger cross-sectional die and a less drastic extrusion process involved to obtain lighter colour than spherical shaped extrudates [42].

Note: *Each parameters values were expressed as (mean± standard deviation; n = 3); indicating the significant differences (p < 0.05) among the sphere and star shaped products considering the interaction between the Millet and die.

Anti-Nutritional Component of Puffs and Extruded Products

The extrusion-cooking process and the type of raw material used can influence the levels of anti-nutritional compounds in legume extrudates [43,31]. A comparison of the native Puffs shown in Table 2 and the extrudates in Table 4 revealed different kind of behaviour for various anti-nutritional compounds. In present study the phytates content decreased as the extrusion-cooking of finger Millet (12% on average) and foxtail Millet (7.9% on average) Puffs initiated for both star-shaped and spherical products, possibly on account of thermal processing that is associated with the extrusion-cooking process. [12] found that total phytates were more greatly reduced in lentil flour extruded at 160°C compared to 140°C. However, oligosaccharides, particularly stachyose and raffinose increased during extrusion cooking. This could be attributed to the high temperature and pressure involved in extrusion-cooking, which break the bonding between oligosaccharides and other macromolecules, or alter the food matrix structure, leading to better extractability of anti-nutritional compounds [6]. Similar kind of results were observed in pea-rice gluten-free expanded products and extruded lentil snacks by other researchers [6,25]. It was observed that there were differences in verbascose, stachyose and raffinose content in Millets extrudates just because of Millet type, die configuration and their interaction, but their phytic acid was not affected by die shape. In case of pearl Millet based spherical extrudates have higher verbascose content than starshaped, while raffinose content increased to 7% in the latter. Common bean-based star extrudates had higher stachyose and raffinose content compared to spheres made from the same flour, which decreased by 1.7% and 7.5%, respectively [23]. Extrusion conditions and legume type affected oligosaccharide behaviour, with a higher temperature and pressure increasing stachyose and raffinose contents. An obtained results showed that oligosaccharides content was found higher in spherical shaped products than the star-shaped after die inducing higher pressure and heat generation, particularly for raffinose [39].

Sensory Evaluation of the Extruded Products

The sensory attributes like appearance, texture, taste, and aftertaste of the extruded products were significantly (p <0.05) affected by the type of flour and die used. The products ranking is decided as per test results (Table 5). As per sensory attributes the star-shaped extrudates prepared from pearl Millet were found to be least preferred acquiring lowest rank sum for “texture” due to their hard structure and difficult to chew, bland taste, and aftertaste. However, pearl Millet based spherical extrudates were liked for “appearance” and “aftertaste”, similarly notified in spherical and star-shaped extrudates from finger and foxtail Millets. Previously [10] studies showed that high values of BD and hardness can produce undesirable products for consumers. In contrast, common Millet extrudates, particularly a spherical one, were preferred in terms of all considered attributes owing to their properly puffed and crunchy nature, pleasant taste and aftertaste as discussed by [28]. In another study, extrudates made from blends of red lentil and corn were found to be more accepted than including (100%) lentil extrudates [17]. In general, the spherical shaped extruded products were favored as compared to star-shaped ones, indicating that preferred shape plays a crucial role in consumer perception and acceptability of food products. This is supported by studies that suggest that shape can even affect taste perception. However, there was no significant difference observed between spherical and star-shaped extrudates in terms of taste and aftertaste, possibly because textural features, such as appearance and structure, had a greater impact.

Note: *Each parameters values were expressed as (mean± standard deviation; n = 3).

Conclusion

The study aimed to understand how the die configuration affects the extrusion of Millets. Most industry dies have a circular cross section, but the effect of a star-shaped die on Millet was still unknown. Obtain results of the study showed that the die-hole diameter significantly affects the physicochemical properties and sensory qualities of the extruded snacks. The use of a star-shaped die to produce products with a lower ER and higher BD than spherical extrudates and it is preferred one on account of lowering the friction resistance during extrusion. The increased knowledge on die configuration could aid in the expansion of Millet-based raw materials at maximal level to meet consumer satisfaction for healthy and palatable food products.

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Journals on Medical Informatics

Nigella Sativa use for the Treatment of Cancer Tasawar

Introduction

Cancer is a big problem in society today and is the second most common cause of death after heart attacks. Every year, many people die from different types of cancer, even though we try really hard to find ways to stop and treat it. In the last 100 years, modern medicine has made big progress in treating diseases. But a lot of illnesses, like different types of cancer, are still not completely treated. Researchers are looking at both old and new ways of healing to find new and effective treatments [1]. Nigella sativa has been used as medicine for a long time. This tradition started in Southeast Asia and then was used in ancient Egypt, Greece, the Middle East, and Africa. The Islamic tradition and the healing power of medicine are important for healing and are an unusual type of medical treatment [2]. This plant is a flower plant and its seeds are used as a spice in cooking. In English, the seed is often called black cumin, and in ancient Latin it was named “Panacea”, showing that it was used for healing. In Arabic, the seed is called “Habbah Sawda” or “Habbat el Baraka”, which means “Seed of blessing”. In Arabic, people call the seed “Habbah Sawda” or “Habbat el Baraka”, which means “Seed of blessing”. This tree is called “Kalo jeera” in Bangladesh, “Kalonji” in India, and “Hak Jung Chou” in China [3]. The plant’s seeds and oils are important for medicine. The main parts of N. Sativa could help keep you healthy and might treat different illnesses like cancer. One way it works well is by lowering the chance of atherosclerosis. It does this by lowering bad cholesterol in the blood and raising good cholesterol. It helps diabetes by making the body healthier and protecting the cells that make insulin in the pancreas. This can help as a treatment for diabetes. Take care of and keep safe. It helps control high blood pressure [4]. It effectively reduces airway inflammation in people with asthma, and its components show potential as a complementary treatment for schistosomiasis. In addition, its oil also protects kidney tissues from damage caused by harmful oxygen molecules, thereby preventing kidney dysfunction and structural abnormalities. For countless centuries, seeds, oils and extracts derived from N. sativa have been used for their anti-cancer properties in traditional systems of medicine such as Unani, Ayurveda and ancient Chinese medicine. These systems were developed in Arabic, Indo-Bangla and Chinese respectively [5].

Role of sativa as Anti-Cancer Mediators

A lot of useful substances have been found in the seeds of N. Sativa is a type of cannabis plant. Instead, it should be written in a simpler and easier to understand language. Nsativa seeds contain oils, proteins, alkaloids and saponins. These components were analyzed to quantify four important pharmacological elements in the oil: thymoquinone, dithymoquinone, thymohydroquinone and thymol. Nsativa seeds are seeds of the sativa plant. The main component in the essential and fixed oils of the seeds, called thymoquinone, is believed to be responsible for a significant portion of their biological effects. Thymoquinone is often recognized for its powerful abilities as an antioxidant, anticancer, and antimutagenic agent. In addition, thymoquinone has acceptable safety levels, especially when administered orally to animals in experiments. Alpha-hederin is a compound found in black seed that comes from the seeds of the N. sativa plant [6].

Blood Cancer

Thymoquinone stops the growth of a type of cancer cells called HL-60 cells, which are found in human myeloid leukemia. Researchers studied different forms of thymoquinone with 6-alkyl residues and terpenes in HL-60 cells and 518A2 melanoma. The scientists discovered that these substances can cause a process called apoptosis, which is when the DNA breaks into pieces, the energy in the cell decreases, and there is a small increase in harmful chemicals. They found that α-hederin made P388 murine leukemia cells die by causing more apoptosis to happen, and this happened more as the dose of α-hederin and the time went up [7].

Breast Cancer

The mix of alfalfa, melatonin and retinoic acid helped lessen the bad effects of DMBA on breast cancer in mice. Thymoquinone was tried on breast cancer cells called MCF-7/Topo. Thymoquinone has a special part called terpene and 6-alkyl. They discovered that these substances made cells die through a process called apoptosis [7].

Colon Cancer

Thymoquinone can help fight colon cancer cells and promote cell death. This was shown in a study where the N.sativa volatile oil was used on colon cancer cell line HCT116. N.sativa can stop the growth of colon cancer in rats, after it has started, without causing any noticeable negative effects. Thymoquinone is a substance that can be used as a treatment for colon cancer cells. It works similarly to a drug called 5-flurouracil. However, thymoquinone did not have any effect on HT-29 (colon adenocarcinoma) cells [1].

Pancreatic Cancer

Thymoquinone is the most important chemical in Nigella sativa, which is known for its important healing properties. The study looked at sativa oil extract affects the way pancreatic cancer cells grow and die. Scientists have suggested using thymoquinone to stop inflammation and encourage cells to die in a certain way [8]. This could be a new way to treat inflammation. Thymoquinone can help make pancreatic tumors more sensitive to standard treatments by reducing the effects of gemcitabine or oxaliplatin on NF-kappa B activation. Mucin 4, a big molecule with sugar attached to it, is not working properly in pancreatic cancer. This strange expression is important for many things like cells change, grow, spread, and resist chemotherapy in pancreatic cancer. The study looked at thymoquinone affects MUC4 expression in pancreatic cancer cells [7].

Hepatic Cancer

A lot of research has been done to study well different treatments can kill cancer cells. This study wanted to see if sativa seeds can affect liver cancer cells grow. The test showed that N can stop HepG2 cells by 88% after being left with them for 24 hours at different amounts. So, using sativa extract is very important in academic discussions. Thymoquinone has been found to help the body’s quinone reductase and glutathione transferase work better when taken by mouth. The properties of thymoquinone make it possible to prevent cancer caused by chemicals and protect the liver from damage [9].

Lung Cancer

The cancer-fighting ability of α-hederin found in N.staiva is being studied. The research looks at sativa affects lung cancer in mice. A study also showed that putting honey and N. Adding sativa to the diet makes a big difference. Sativa helps protect against oxidative stress and inflammation caused by certain chemicals and can help prevent lung, skin, and colon cancer. Alfalfa has been looked at a lot because it might have healing powers [2]. These chemicals have different effects on living things, such as reducing inflammation, fighting cancer, and preventing damage caused by harmful substances. Additionally, they look like they could help treat heart problems and might assist in cancer treatment. Understanding α-hederin and thymoquinone work can help us use them in medicine. More research is needed to understand they can help people and make sure they are safe and work well in medical treatment. N sativa does not have a big impact on cell death or programmed cell death in lung and laryngeal cancer cells [10].

Skin Cancer

Topical use of N. sativa is a common form of management used in a variety of settings. sativa extract showed an inhibitory effect on the initiation and promotion stages of skin carcinogenesis in mice when administered intraperitoneally. Skin application of 20-methylcholanthrene resulted in a significant reduction in soft tissue sarcomas, which was limited to 33. 3%, compared to 100% incidence observed in the MCA control group [11].

Fibrosarcoma

Thymoquinone, taken from the seeds of Nigella sativa, has been studied a lot for its possible healing powers because of its different biological effects. The administration of sativa one week prior to and following MCA treatment exhibited a notable hindrance in the development of fibrosarcoma tumor occurrences, as well as a reduction in tumor mass, by 43% and 34% respectively, in comparison to the outcomes observed in the MCA solitary treatment group. Furthermore, it was observed that thymoquinone exhibited a delayed onset of fibrosarcoma tumors induced by the administration of MCA. Moreover, in vitro investigations demonstrated that thymoquinone exhibited inhibitory effects on the viability of fibrosarcoma cells. Alfalfa oil is a special kind of oil from the Medicago plant that can decrease the ability of human fibrosarcoma cells to break down fibrin in lab tests [1].

Renal Cancer

There is an existing body of research that highlights the potential chemo-preventive efficacy of N.sativa, a substance that has garnered significant scientific interest. N sativa has inhibitory effects on renal oxidative stress, hyperproliferative response, and iron nitrilotriacetate-induced renal carcinogenesis. In this experimental study, alfalfa was administered orally to rats for therapeutic purposes.The administration of sativa elicited a pronounced reduction in the generation of H2O2, synthesis of DNA, and occurrence of tumors [12].

Prostate Cancer

Thymoquinone comes from Nigella seeds and has many important medical qualities. The extract from the sativa plant can slow down the making of DNA, stop cells from growing, and reduce the ability of cancer cells in the prostate to survive. These changes were seen only in cells that have cancer, not in cells that do not have cancer. It was found that this result happened because the androgen receptor and transcription factor were decreased [13]. Thymoquinone was found to work well in treating both hormone sensitive and hormone-refractory prostate cancer in different experiments. Research done in the lab and in animals shows that thymoquinone can stop the growth of new blood vessels. In addition, thymoquinone was found to stop the growth of blood vessels in a human prostate cancer model in mice. Also, when used in small amounts, thymoquinone stops the growth of human prostate tumors, with very few chemical side effects. Furthermore, thymoquinone affects endothelial cells more than cancer cells by causing cell death, stopping cell growth, and blocking cell movement. Thymoquinone stopped the activation of a protein called extracellular signal-regulated kinase, which is usually turned on by a substance called vascular endothelial growth factor. However, it did not stop the activation of the vascular endothelial growth factor receptor 2 [14].

Cervical Cancer

The extracts of N. sativa were obtained using methanol, n-hexane, and chloroform. The sativa plant caused human cervical cancer cells to die. “We looked at terpene-terminated 6-alkylthymoquinone residues affect cervical cancer cells that are resistant to multiple drugs. ” Thymoquinone derivatives have been discovered to make cells die in a certain way called apoptosis [4].

Molecular Mechanism of sativa Action Against Cancer

Cancer happens when cells grow in an unusual way because of changes in the genes. So, any medicine that fights cancer can either protect DNA from changing or kill cancer cells that have changed. N.sativa seeds have been extensively studied for their pharmacological properties. Thymoquinone, a major active compound found in N. Sativa seeds, has demonstrated promising therapeutic effects in numerous medical conditions. These studies have shed light on the potential of thymoquinone as a valuable agent in disease prevention and treatment. Sativa exhibits its efficacy in combatting cancer cells through various molecular pathways. Possible mechanisms underlying the action of thymoquinone. Thymoquinone helps kill cancer cells by turning on genes that cause cell death and turning off genes that keep the cells alive. Thymoquinone effectively hinders the activation of Akt by means of dephosphorylation, ultimately impeding the viability of cancerous cells [15]. There is currently ongoing research regarding topic N.sativa in the academic community. Nsativa or thymoquinone oil exhibits antioxidant properties, leading to enhanced enzymatic activity of antioxidant enzymes including superoxide dismutase, catalase and glutathione peroxidase. Notably, increased activity of these antioxidant enzymes has been shown to be beneficial in fighting various forms of cancer. The use of alfalfa oil or thymoquinone has been observed to reduce the toxicity of various anticancer drugs due to enhanced activation of antioxidant mechanisms. This result suggests that these drugs have great promise for clinical use in mitigating the harmful effects associated with anticancer drugs [4].

Concluding rrmarkers

Nigella sativa (N.sativa), commonly known as dim seeds, have been broadly inspected for their promising anti-cancer properties. Many in vitro and in vivo tests have outlined the practicality of N.sativa in preventing the improvement and development of diverse sorts of cancer cells. In extension to its facilitate cytotoxic impacts, N.sativa has been found to have solid antioxidant, anti-inflammatory, and immunomodulatory properties, all of which contribute to its anti-cancer development. Besides, N.sativa has appeared potential in sensitizing cancer cells to customary chemotherapy and diminishing chemotherapy-induced side impacts [16].

Nigella Sativa Good for Cancer

Nigella sativa has attracted a lot of interest from researchers and scientists. Extracts and seeds of the N. sativa plant and its active ingredient thymoquinone have been thoroughly studied, with excellent results showing that N. sativa has medicinal potential. The sativa strain tends to have medicinal properties that can be effective in treating a variety of ailments, including cancer [7].

Sativa Extracts be used to Treat Cancer

N.sativa extracts have the prospective application in the advancement of efficacious therapeutic agents for combating cancer. These fractions can act alone or in combination with chemotherapeutic drugs that have proven to be effective agents for modulating tumor initiation, proliferation and metastasis, making them possible treatments for many types of cancer [17].

Pro-Apoptotic and Anti-Proliferative Effects of Sativa

N.sativa has the ability to fight against cancer. Researchers have gathered a lot of evidence about sativa by studying it outside of living organisms and inside them. They have used different types of cells and animals to do this. The scientists in the study said that they didn’t look at the extracts from each individual plant in the mixture could fight cancer because only the mixture is used in cancer treatment. Sativa extracts are extracts from the sativa plant. In an initial test on living organisms, we applied N.sativa using a cream or ointment on the surface. The N.sativa extract slowed down the development of skin cancer and reduced the appearance in mice when they were exposed to certain chemicals [18].

Signaling Pathways Fundamental the Anti-Cancer Effects of Sativa

Many tests were done in the lab and on living organisms to understand N.sativa fights against cancer at a molecular and cellular level. Sativa is a type of plant. The main ways that N.sativa (a specific substance) helps fight against cancer are not yet fully understood, but they have been well-documented. The effects of sativa are mostly due to their capability to control the action of important enzymes. Reduce swelling and encourage the natural death of cancer cells [15].

Sativa Phytoconstituents and Anti-Cancer Effects

The anti-cancer impacts of N.sativa are exceptionally critical. The most fixing in N.sativa, called thymoquinone, has been connected to its impacts. Thymoquinone has been found to have a few useful impacts on cancer cells. It can offer assistance halt their development, empower cell passing, secure against harm caused by substances called oxidants, decrease the probability of changes, anticipate the arrangement of modern blood vessels that tumors got to develop, and moderate down the spread of cancer cells to other parts of the body. This N.sativa herb has been appeared to have properties that can battle cancer and murder cells. Be that as it may, we do not completely get it it works however, so more investigate is needed to figure out the points of interest. Sativa phytoconstituents are the common compounds found within the sativa plant (Figure 1) [4].

Figure 1

Conclusion

Nsativa is a very popular herb that has been used by people for a long time. Many people think Nsativa is a special plant that can help heal and reduce the effects of infections, such as cancer. The ability of N.staiva to fight cancer. Sativa is good because it can help stop cells from growing, help cells die, and protect cells from damage and cancer. N. sativa ability to defend effectively. Sativa can help stop tumors from forming and spreading, partly because it can prevent them from getting worse and has a mild stimulating effect that is safe. The tests done in the lab and on living organisms show that N. Sativa extract can be used to create helpful and powerful starting materials that can be used at different times during the process. Cancer can form tumors in different parts of the body. There are different treatments for different types of cancer. More testing is needed to understand N helps fight cancer at the atomic and cellular level. Sativa researchers want to understand the exact ways that N. stops certain signals in the body. The sativa extract is involved in the development of tumors and cancer. In the future, we need to study N.staiva can work together with other things to fight cancer. Sativa extract being used to prevent and treat cancer in research and medical settings.

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Journals on Cancer Medicine

Nigella Sativa use for the Treatment of Cancer Tasawar

Introduction

Role of sativa as Anti-Cancer Mediators

Blood Cancer

Breast Cancer

Colon Cancer

Pancreatic Cancer

Hepatic Cancer

Lung Cancer

Skin Cancer

Fibrosarcoma

Renal Cancer

Prostate Cancer

Cervical Cancer

Molecular Mechanism of sativa Action Against Cancer

Concluding rrmarkers

Nigella Sativa Good for Cancer

Sativa Extracts be used to Treat Cancer

Pro-Apoptotic and Anti-Proliferative Effects of Sativa

Signaling Pathways Fundamental the Anti-Cancer Effects of Sativa

Sativa Phytoconstituents and Anti-Cancer Effects

Figure 1

Conclusion

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Medical and Medicinal Journal

The Review about the Kinetics of Monoterpenes Formation in Plants of Sort Artemisia

Essential oils from plants of sort Artemisia are applicable in various branches of perfumery, pharmaceutical and the food-processing industry. Predominant components and their content define practical value of essential oil. Many explorers [1-3] studied the oils composition obtained from plants of sort Artemisia. The basic valuable components of essential oils are various representatives of monoterpenes. The huge actual data about the monoterpenic reproduction during the vegetative period of various ethereous-carrier plants is saved up. But numerous researches, as a rule, confine some selves by the only exposition of exact predominant components within the limits of any taxon-carrier for practical purposes [4-6]. In the present review the author’s kinetic model affixed to the reproduction of monoterpenic hydrocarbons in life cycle of five Crimean plants of sort Artemisia [7-9] and to that from Eastern Siberia (A.glauca) [10] is used. Monoterpenes possess various biological effect: one with an open chain are used in perfumery (citral, geraniol, linalyl ester) and other are used in the food-processing industry as gustatory components. Between cyclical monoterpenes there are widely known medical products: menthol, terpin hydrate (monocyclic), camphor (bicyclic) [11-13]. Composition of monoterpenic components and their content in plants define as the quality and the industrial utility of received essential oil. By the method of hydro distilling from plants of type Artemisia abrotanum, A.annua, A.dracunculus, A.glauca, A.taurica, A.scoparia essential oils had been received, and then they are parted by the chromate-mass spectroscopy on components and are identified.

Then monoterpenic components of essential oils have been distributed according to their ring formation degree. For each degree of ring formation, as shown in the article [14], one calculated the normalized concentrations according to vegetation phases, namely: I – the vegetation beginning, II – budding, III – mass blooming, IV – maturing of seeds. Thus, one obtained the accumulation mass fractions of not exact compounds in studied plants, but the generalized carbocyclic structures, such as: acyclic (X2), monocyclic (Y2) and bicyclic (Z) (Figure 1), which can be extrapolated on all plants and to evolve the general regularities of their accumulation. The finding of singularities in a connatural monoterpenes biosynthesis was made by own kinetic model reduced in the article [15,16]. Data about the qualitative composition and the quantitative maintenance of monoterpene’s components in the essential oils obtained from plants of sort Artemisia balchanorum Krasch was given in the article [15]. Such experimental data about the essential oil’s composition have been compared with the steps of calculated curves so that these curves must be maximum corresponded to the data (Figure 1) according to the known algorithm of the regressive analysis (Fisher’s criterion), reduced in the articles [15,16]. The received graphic solutions of a differential equations system represent those selves a data bank of monoterpenes accumulative dynamics in the plants growing as in the Crimean region and in Eastern Siberia also. Thus, the quality and quantitative content of monoterpene’s hydrocarbons in two sort plants samples of Artemisia balchanorum is detected during all phases of vegetation with the distribution of the components according to the degree of cyclization and with the definition of their group quantity properties and it is placed on the (Figure 1).

Also, we constructed the similar calculated curves for the extract compositions from some other plants samples of vegetable species Artemisia balchanorum according to their ring formation degree during all phase’s vegetation (the phases I-IV), as it is demonstrated on the (Figure 2). The steps of graphic curves (1) (Figures 1 & 2) indicate on the possibility to describe vegetative types by the kinetic methods according to the graphic maps, which demonstrate the efficiency of the investigated monoterpenes groups. There are accurate enough graphic differences between six types of the plants, despite the uniform mechanism of biosynthetic process (Figure 2). The constants of biosynthesis and the factor planes for each vegetative type are presented in the article [15]. For the medical industry it is more perspective to cultivate just the plant A. glauca Pall. ex Willd, which is rich monocyclic terpenes (β -fellandren, γ-terpinen, terpinolene) for use that as medical products [10-13]. There is a wide spectrum of benefit performance biological properties [17], which is corresponded to the components of essential oils from plants A. annua L., such as A. abrotanum L., A. dracunculus L., A. scoparia Waldst. et Kit. Artemisia Taurica Willd. Data of Figures 1 & 2 show a possibility to organize the harvesting of these plants such way that its results will lead to demanded composition of monoterpenes in the essential oils according to the corresponded vegetative phase.

One chooses some perspective sort-samples of these plants, which are used now as the agronomic culture for a pharmacology. For example: the sort Beginner in the type of A.annua L. is rich acyclic components, namely: 1,8-tsineol, Artemisia ketone, Artemisia alcohol having a high quantity of bicyclic camphor ketone; the sort Evksin in the type of A.abrotanum L. contains 1,8-tsineol and α -thujone as principal components of essential oil, is designed to be used not only in a medicine, but also in a perfumery, and also in the food-processing industry; one made two sorts named as WIM and the Crystal in the type of A.dracunculus L., which contain principal components, such as linalyl ester (at one) and neril-acetate (at another); the sort Branching in the type of A.scoparia Waldst.et Kit contains a high quantity of capillene in the essential oil; the sort Alupka in the type of A.taurica Willd contains bicyclic monoterpenes (thujones are the principal components and the camphor is presented also) [18]. The received outcomes about a monoterpenic hydrocarbons reproduction during the life cycle of ethereous-carrier plants will form a data bank of qualitative and quantitative composition of monoterpenic components in the essential oils from five Crimean plants types of sort Artemisia and one of that from Eastern Siberia. It is offered performances of vegetative types using the graphic maps of their efficiency according to degree of ring monoterpenes formation. It is offered to use the considered data bank at a choice of the plants sort-examples to cultivate them for exact industrial needs, to plane the terms of plants harvesting on purpose to extract from them the essential oil possessed by the known properties, to do a forecasting and to study the singularities of plants vegetation in agricultural region and to plane the compositions from the oils for industrial needs [19].

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Top Medical Open Access Journal

Buffalo Milk Adulteration in South Punjab (Pakistan) -A Public Health Hazard

Buffalo milk, one of the substantial known human foods, provides many nutrients such as carbohydrates, fats, proteins, vitamins, minerals, and other important components (Yasmin et al., 2012). Buffalo milk is popular in Pakistan and other south Asian countries. Buffalo milk is unique due to less cholesterol, high fat, content more calories that add to its health benefits. Moreover, it is a good diet for developing healthy bones, maintaining dental health, and preventing cardiovascular disorders. Buffalo milk is thick and creamy in consistency therefore suitable for manufacturing traditional dairy products including yogurt, cottage cheese (paneer) and traditional milk products like ghee and khoa (Haque [1]). According to authors, shorter summers (hence shorter periods of pasture grazing) in the Nordic countries could explain CLA differences between the Nordic and other European countries. However, it was not known if such changes would occur under tropical conditions with the traditional feeding system of Sindh, (Pakistan), where animals are fed on forages throughout the year, except in winter and late spring when shortage of green forage occurs, and ruminants are fed crops by product along with available green fodder. Pakistan ranks as the 5th largest producer of milk in the world with over 26 million buffaloes, 56 million sheep, 24 million of each cattle and goats. Buffaloes are the major milk producing animal, accounting for about 75% of all milk produced. They are concentrated in irrigation areas and long rivers, as are the human population.

Goat and ewe milk are not proceeded for commercial purposes in Sindh (Pakistan), and these animals are mostly reared for meat purpose, while milking is a secondary function, which provide extra source of income to poor farmers (Sarwar et al., 2002). However, data on milk fatty acid composition from ruminant species from Pakistan is scarce. Milk adulteration became a global concern after breakthrough of melamine adulteration in Chinese infant milk formula. Worldwide, it is a terrible situation that milk is being very easily adulterated and the situation is significantly worse in underdeveloped countries due to lack of adequate monitoring and absence of proper law enforcement (Xin [2]). Dairy Milk adulteration is a socioeconomic issue in developing countries. The act of adding adulterants makes it unfit for human consumption and dairy products fail to meet the legal standards. It brings not only brings unfavorable consequences in terms of major economic losses for the processing industry, but also a major health hazards for the consumers from infants to adults worldwide. The consumption of adulterated milk causes several critical health disorders including dysentery, colon ulcers, nephrosis, disturbance of cardiovascular system (Hanford [3]). In Pakistan limited studies are available on level of dairy milk adulteration, a major public health hazard. Limited studies have been conducted on buffalo milk composition variation in different seasons but none in south Punjab where summer season lasts for long duration and winter lasts for short period.

The first objective of the study was to estimate variation in composition of buffalo milk. The second objective was to know about adulterant types used in buffalo milk supplied for human consumption in South Punjab region of the country.

Methodology

A total of 100 milk samples were collected by convenience sampling method from various cities in southern Punjab. Milk samples were collected in two seasons. Winter(n=50) sampling was conducted from December to March and summer(n=50) sampling was conducted from April to June from same cities. Milk samples were collected from dairy farms(n=40), local vendors(n=30), and commercial chillers(n=30). The raw milk samples were collected from homogenized milk into sterilized Falcon tubes to avoid any type of contamination. All milk samples were labelled and placed in ice filled cooler box. The samples were transported to the Milk Testing Laboratory Punjab Food Authority, Multan and preserved at 4-8°C until analysis. For estimation of milk fat, protein, lactose, and total solids Lactoscan milk analyzer was used (Miltonic Ltd.series7035, Bulgaria). For detection of adulterants (starch, urea, hypochlorite, pulverized soap, formalin, sugar, skim milk, boric acid and detergent) user friendly Latte Adulterazione kit was used according to the manufacturer’s protocol. The data generated was entered in software SPSS and independent t test was applied to find significant difference in milk composition.

Results

In the present study, change in milk composition during extreme weather conditions of South Punjab region were investigated. Data shows that fat content of milk samples was significantly (P<0.05) different in winter and summer seasons. Milk protein content was also significantly(P<0.05) different in winter and summer season. Milk lactose content was non significantly (P>0.05) different in winter and summer seasons. Milk total solids content showed significant (P<0.05) difference in winter and summer season as shown in Table 1. In the present study data from dairy farms showed no milk adulteration in winter and summer season in South Punjab region. Milk samples acquired from local vendors showed use of skim milk followed by detergent as adulterants were high both in winter (52.5%;33.33%) and summer (72%;31.33%) seasons. Milk samples acquired from commercial chillers showed use of skim milk followed by starch as adulterants were high both in winter (60%; 33.33%) and summer season (60%; 33.33%). The use of urea as milk adulterant was found in summer (28.3%) in buffalo milk samples as shown in Table 2.

Note: P≤0.05 (*shows that value is significant)

Discussion

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Out of total dairy milk produced in Pakistan, buffalo contributes about 68 %, followed by other dairy animals. Due to the high fat contents of buffalo milk, it is the most preferred by people among other dairy products. In the present study buffalo milk samples showed difference in fat, protein, and total solids contents in winter and summer seasons. Saadi, et al. (2019) finding is in line with our study and show significant difference (p<0.001) with highest percentage of fat 4.48 % in milk in winter while values decrease to 2.95% in summer. The decrease in fat in summer may be due to the length of light compared to dark. The higher the ratio between light and dark the lower the proportion of fat due to increased prolactin secretion whose concentration in plasma is higher in summer than in winter (Ozrenck [4]). As for the effect of nutrition the basis in the composition of fat depended on the composition of acetate in the rumen. Food such as grain feeding that reduces acetate production will also reduce fat concentration in milk produced during the summer (Vildirim and Cimen, 2009). Percentage of protein was highest in milk produced from cows, which fed on concentrated feeds in the winter and amounted to 3.46% and decreased to 2.93% for the milk of cows that fed on herbs in the summer, and attributed the reason for high protein in the winter to the diet content high in protein while in the summer the nutrition was low in protein content (Colombari [5]). The low percentage of fiber in cattle (concentrated feed) resulted in increased protein content in milk produced. On the contrary, when they fed on green the protein decreased (Petitclerc, et al. 2000). Significant differences were found in the percentage of total solids, as the proportion of fat increased the proportion of total solids (Pavel [6]). The freezing point and lactose sugar showed no significant difference. Difference in milk composition in both seasons might be due to change in feed and water consumption by buffaloes (Afzal [7,8]). In the present study buffalo milk adulteration was observed in both seasons. Skim milk powder was the most used adulterant for thickening of milk in the study area as reported in literature (Azad [9]). Urea added in buffalo milk to increase non protein nitrogen content. Urea adulteration has carcinogenic effects on human health. Starch is used as a thickening agent in milk. Starch adulteration can cause diarrhea and its accumulation in the body might be fatal for diabetic patients (Yadav [10]). Detergents are added to emulsify and dissolve the oil in water to give frothy solution, a desired characteristic of milk. Consumption of adulterated milk leads to kidney failure, gastritis, and intestinal inflammation (Rennie [11-14]).

Conclusion

People living in developing countries like Pakistan, which is already far behind in delivering health services, are deprived of pure healthy buffalo milk. The adulterated milk is of low quality and responsible for introducing hazardous substances leading to serious health hazards in consumers. Therefore, it is need of the hour to devise an efficient and reliable quality control system that will regularly monitor the activities of malpractices in the dairy industry. It is the responsibility of the government to formulate an effective strategy to ensure the access of fresh and quality raw milk to people so that infants and adults both can enjoy a healthy life by having pure natural milk. General public awareness, effective monitoring measures and regulatory system for quality control of milk and dairy products can play a crucial role in minimizing milk adulteration.

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