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Effect of Feed on the Mineral Composition of Labeo Rohita

Khalid Javed Iqbal*1, Muhammad Ashraf1, Arshad Javid2, Farzana Abbas1, Muhammad Hafeez-ur-Rehman1, Fayyaz Rasool1, Noor Khan1 , Sumaira Abbas1 and Muhammad Altaf 2
ABSTRACT
Studies were conducted to evaluate the effect of plant-fishmeal feed and/or plant by-product based feed on minerals composition of Labeo rohita. Fish fed on rice polish alone served as control (T0). Feed ingredients were grouped together with two ingredients in each test diet which served as an independent trial during these studies. Group 1(T1) contained guar meal and canola meal, group 2(T2) soybean meal and cotton seed meal, group 3(T3) guar meal and cotton seed meal, group 4(T4) soybean meal and canola meal and group 5(T5) fishmeal and canola meal. Each group including control had two replicates. 12 earthen ponds with uniform area of 0.03 ha each, were randomly stocked with 100 fish (average weight 200 g) in each following standard stocking protocols. All the 12 ponds were then randomly allotted to individual treatment including control group. Experimental fish were fed @ 4% of their wet biomass twice a day. Minerals specifically Na, Ca, Fe, Zn, and Cu significantly differed (P?0.05) among treatments which might be linked with their variable release in digestive system of fish in the presence of various anti-nutritional factors.
Key Words: fishmeal; soybean meal; canola meal; Ca; Na.
INTRODUCTION Fish is rich in animal protein, low in cholesterol and high in unsaturated fatty acids (Kromhout et al., 1995; Zenebe et al., 1998a; Arts et al., 2001; Fawole et al., 2007) and due its these peculiar qualities is preferred over red meats (Sadiku and Oladimeji, 1991; Mozaffarian et al., 2003; Foran et al., 2005;). Nutritional quality of fish is however, not uniform and varies a lot among different fish species even within species when cultured under environments and different culture systems. Among herbivorous fish varieties Labeo rohita is preferred among consumers due to its typical taste and texture and among culturists due to growth, hardiness and wide range feeding habits. That is the reason that it is dominant fish in current fish cultural practices (Khan et al., 2004; Hussain et al., 2011; FAO, 2000; Chaudhuri et al., 1974).
Other than nutritional competencies the fish is an important economic source, and its culture is rapidly growing not only in developing countries but in developed contraries too (Delgado et al., 2002; Louka et al., 2004). The success of fish culture depends on availability and selection of appropriate diets that are proficiently digested, are cost effective and provide the necessary nutrients for optimal growth (Mokolensang et al., 2003). Improvement and selection of appropriate feed ingredients has pronounced effect on the nutritional values, fish growth and its adjunct qualities (Shioya et al., 2011; Yang et al., 2011). Cost effective quality feed has pivotal role in fish production and has always been a constraint in the expansion of fish culture and in sustained development of aquaculture industry. It determines growth, flesh composition, especially lipid, mineral content of produced fish and ultimately market response (Izquierdo et al., 2003; Rasmussen, 2001).
Among other nutrients minerals also has an important role and contribute to the growth of fish being an integral components of many enzymes involved metabolism (Glover and Hogstrand, 2002). Several minerals are required for proper development and normal execution of organism’s bodily functions as Ca is necessary element for the bone development (Erkan and Ozden, 2007) and Ca, Mg, Na and K, are involved in cellular metabolism which are usually found in higher quantities in biological tissues (Wagner and Boman, 2003). Zn is well known to be involved in most metabolic pathways in plants and animals (Hambidge, 2000). Copper, iron and manganese are essential for maintenance of normal growth and reproduction (Turkmen et al., 2005; Roy and Lall, 2006). Fish is a major source of Fe (Fraga, 2005) which is involved in blood synthesis in liver (Wagner and Boman, 2003), is an integral component of oxygen carrying protein from lungs to the tissues (Wagner and Boman, 2003; Camara et al., 2005). Mn is required in minute quantities on daily basis for better health and growth in humans and its deficiency may result in nervous system disorder (Agency for Toxic Substances and Disease Registry, 2004). Keeping in view all the above mentioned concerns the present study is planned to find out the effect of plant-fishmeal feed and/or plant by-product based feed on minerals profile of Labeo rohita.
MATERIALS AND METHODS Experimental site and study trials
This three month study was conducted in earthen ponds of the Department of Fisheries and Aquaculture, University of Veterinary and Animal Sciences, Ravi Campus Pattoki, using juvenile Labeo rohita as an experimental animal.
Experimental design
Studies were designed following Completely Randomized Design (CRD). There were 5 treatments and a control with two replicates in each group and whole trial was managed in 12 ponds. 100 juveniles of Labeo rohita having mean body weight of 200g were randomly stocked in each pond (0.03 ha) and then all these ponds were arbitrarily distributed among 5 treatments and a control. Five experimental diets pertaining to each treatment, by proportionate ratio of the two feed ingredients was maintained at 1:1 i.e. (T1) guar meal and canola meal, (T2) soybean meal and cotton seed meal, (T3) guar meal and cotton seed meal, (T4) soybean meal and canola meal, (T5) fishmeal and canola meal and a control diet (T0) i.e. rice polish with two replicates in each. Fish were regularly fed @ 4% of wet body weight twice a day.
Proximate analysis
Feed proximate analysis was analyzed by using Büchi NIR Technology (Büchi NIRFlex N-500) Feed were dried and finally ground in pestle and mortar and then placed in sampler cups. The cups were placed in Büchi NIR machine for two minutes which then displayed a complete proximate analysis report which was saved for future use (Table 1).
Table 1 Proximate analysis of feed combinations
Analysis
T1
T2
T3
T4
T5
T0
Protein %
36.76±1.20
38.45±2.29
37.64±2.33
37.56±1.78
40.35±3.02
6.07±0.64
Moisture %
7.08±1.31
9.68±2.11
7.135±.188
9.62±2.26
7.33±1.79
4.92±1.21
Fat %
1.77±0.55
1.42±0.54
1.60±0.87
1.35±0.65
4.87±1.48
3.15±0.29
Ash%
8.23±0.39
12.48±2.13
12.35±2.44
8.35±2.38
15.59±3.49
6.30±1.27
Kcal/g
4.09±1.11
4.07±1.37
4.08±0.89
4.06±1.05
4.25±1.91
4.16±2.01
Mineral analysis
Well ground 0.5 g sample was taken in conical flask which 10 ml HNO3 was added in. Mixture was then boiled for 15 minutes at 60 0C and then 5 ml perchloric acid was added and boiled it again for another 15 minutes at 60 0C. Sample flask was then placed on hot plate and heated till sample volume reduced to 1 ml. This sample was diluted to 100 ml by addition of distilled water. Sodium (Na) and potassium (K) were measured by flame photometric method while calcium (Ca), Iron (Fe), zinc (Zn), copper (Cu) and magnesium (Mg) were determined by an atomic absorption spectrophotometer.
Statistical analysis
The data generated during the course of this trial from various sources was analyzed by one way ANOVA using SAS software to determine the significance of various treatment groups. Difference among various means obtained from computation of treatment data sets was compared by Duncan’s Multiple Range Test to indentify the presence of variations. Probability level for these tests was fixed at P?0.05.
RESULTS Mineral composition of Labeo rohita showed statistically significant (P ? 0.05) differences in Na, Ca, Fe, Zn and Cu content while non-significant in K and Mg. Significantly higher Na (27.400±0.98 ppm) was observed in fish fed on T2 and the lowest (18.05±5.30 ppm) in T4, similarly significantly higher Ca content was observed in fish fed on T5 (14.245±0.09 ppm) while the lowest in T3 (10.515±0.09 ppm), significantly higher Fe content was recorded for fish fed on T5 (5.960±0.87 ppm) while the lowest in T4 (1.910±0.14 ppm), significantly higher (0.815±0.09 ppm) and lower (0.470±0.04 ppm) Zn contents were observed in T1 and T2. Higher Cu concentrations were recorded in fish fed on T3 (0.045±0.01 ppm) and lower in fish fed on T0 (0.015±0.01 ppm), higher values of K was observed on T2 (68.550±23.97 ppm) while lower for T3 (53.100±2.82 ppm), maximum Mg values were observed for T1 (3.270±0.11 ppm) and minimum for T2 (2.915±0.10 ppm) (Table -2).
Table 2 Effect of feed on mineral composition of Labeo rohita
Minerals
T1
T2
T3
T4
T5
T0
Na (ppm)
24.600±3.54ab
27.400±0.98a
20.800±0.28ab
18.05±5.30b
21.300±0.28ab
23.700±0.42ab
Ca (ppm)
11.870±0.07d
13.240±0.08b
10.515±0.09e
13.145±0.11bc
14.245±0.09a
12.415±0.76dc
Fe (ppm)
4.690±0.11b
2.465±0.80c
2.535±0.15c
1.910±0.14c
5.960±0.87a
5.010±0.14b
Zn (ppm)
0.815±0.09a
0.470±0.04ab
0.680±0.05ab
0.735±0.05ab
0.700±0.06ab
0.620±0.06b
Cu (ppm)
0.025±0.01bc
0.035±0.01ab
0.045±0.01a
0.025±0.01bc
0.035±0.01ab
0.015±0.01c
Mg(ppm)
3.270±0.11a
2.915±0.10 a
3.035±0.15a
3.095±0.13a
3.015±0.19a
3.125±0.19a
K (ppm)
46.400±13.43a
68.550±23.97a
39.150±0.21a
51.300±11.59a
54.950±0.21a
65.350±1.20a
DISCUSSION In present study mineral composition of Labeo rohita showed significant (P ? 0.05) variation in Na, Ca, Fe, Zn and Cu content in different treatments. Statistically significantly higher Na, Ca, Fe, Zn, Cu were observed in fish were observed in T2, T5, T5, T1, T3 while non-significantly higher K and Mg were observed in T2 and T1 respectively. Contrary to our study Khan et al. (2012) observed non-significant variations in mineral content in major carps reared in mono and polyculture systems. Similarly Luczynska et al. (2009) also observed non-significant differences in fishes having different feeding niches. During present study significantly higher Na was observed in fish fed T2 and lower in fish fed wit T4. Comparatively higher Na concentrations were observed in marine fish by Pirestani et al. (2009). Significantly higher Ca was observed in fish fed on T5 and lower for fish fed on T3. Our findings are in line with Babalola, et al. (2011) who observed significant variations in concentration of Ca among different commercial fish species of Nigeria.
Significantly higher Fe content was observed in fish fed on T5 and lower in fish fed on T4 during present analysis. Mean Fe values were observed within the ranges given by Pirestani et al. (2009) in C. carpio. Babalola et al. (2011) observed non significant differences in Fe content among different commercial fishes of Nigeria. Significantly higher Zn was determined in fish fed with T1 while lower on T2 during present study. Contrary to our study Stezycka et al. (2003) observed higher Zn content in non-predatory fishes and marine fish species (Pirestani et al., 2009). During present study significantly higher Cu concentration was determined in fish, fed on T3 while lower for T0 diet. Contrary to our study Pirestani et al. (2009) observed higher Cu concentrations in fish collected from South Caspian Sea. During present analysis higher Mg concentrations were observed in fish fed on T1 while lower for T2 diets. Our findings confirm Babalola et al. (2011) who observed non-significant differences in Mg content among different commercial fishes of Nigeria. Contrary to our observations Pirestani et al. (2009) found significantly higher Mg content in different commercial fishes of Nigeria. During present study higher K values were determined for fish fed on T2 while lower for T3. Contrary to our study significantly higher K contents were recorded in different commercial fishes of Nigeria (Pirestani et al. 2009), freshwater fish species (Achionye-Nzeh et al. 2011) and commercial fishes of Sudan (Mohamed et al. 2010).
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Investigation of Aspirin Overdose Using Salicylate Assay

Introduction
Aspirin is the most widely used over-the-counter drug in the world. The average tablet contains about 325 milligrams of acetylsalicylic. Aspirin is used to relieve pain, reduce inflammation, and lower fever. Aspirin originally was derived by boiling the bark of the white willow tree. Although the salicin in willow bark has palliative properties, purified salicylic acid was bitter and irritating when taken orally. Phenylsalicylate could be produced by modifying Salicylic acid which resulted in better tasting and less irritating outcomes. Felix Hoffman and Arthur Eichengrün first produced the active ingredient in aspirin, acetylsalicylic acid, in 1893. However, Hippocrates wrote about a bitter powder extracted from willow bark that could ease aches and pains and reduce fevers during the fifth century B.C
200 ml well mixed stomach contents (Sample A)- Transparent Yellow
200 ml hydrolysed stomach contents (Sample B)- Purple
200 ml urine (Sample D)- Light Purple
200 ml negative control (water)- Transparent Yellow
200 ml positive control (400 mg /ml sodium salicylate)- Purple
Solution i and iv which where stomach content and negative control turned Transparent Yellow this means, No change, Negative test. Solutions ii and v turned purple which means that the test where portative for high aspirin traces.
Solution iii turned Light Purple which means that slight trances of aspirin are shown the sample.
– Salicylic acid is a weak acid, and very little as it’s ionized in the stomach after oral consumption. Acetylsalicylic acid is poorly soluble in the acidic conditions of the stomach, which can setback amalgamation of high doses for 8 to 24 hours.
In addition to the increased pH of the small intestine, aspirin is rapidly absorbed due to the increased surface area, which results in allowing more of the salicylate to dissolve. However, aspirin is absorbed much more slowly during overdose, and plasma concentrations can continue to rise for up to 24 hours after ingestion.
As much as 80% of therapeutic doses of salicylic acid are metabolized in the liver.
Renal excretion of salicylic acid becomes ever more important as the metabolic pathways become saturated, because it is extremely responsive to changes in urinary pH. There is a 10 to 20 fold increase in renal clearance when urine pH is increased from 5 to 8. The use of urinary alkalinization utilizes this particular aspect of salicylate elimination.
From the deductions that can be from by results and investigations, it shows that aspirin is absorbed via the small intestine, which then transported to the blood serum. Waste product is transported to urea and disposed via renal excretion. Therefore more aspirin is present in the serum, that why forms a precipitate and the urine dose not as much of it is waste product.
– The acutely toxic dose of aspirin is commonly regard as greater than 150 mg per kg of body mass. Moderate toxicity occurs at doses up to 300 mg/kg, severe toxicity occurs between 300 to 500 mg/kg, and a potentially lethal dose is greater than 500 mg/kg. Chronic toxicity may arise subsequently to doses of 100 mg/kg per day for two or more days.
Around 3 grams per day in divided doses for rheumatoid arthritis is recommended.
Serum salicylate levels may be useful in guiding therapeutic decisions regarding dosage. Serum salicylate levels of 150 to 300 mcg/mL are associated with anti-inflammatory response. However, the incidence of toxicity increases with salicylate levels greater than 200 mcg/mL.
This patients salicylate concentration level levels where 400 mcg/mL. Therefore I believe this person was overdosing on aspirin.
– The key objectionable side effects of aspirin are tinnitus, gastrointestinal ulcers and stomach bleeding contestably in higher doses; this is due to aspirin irritating the stomach lining. In youngsters, aspirin is no longer used to control flu-like symptoms or the symptoms of other viral illnesses, because of the risk of Reye’s syndrome. Another common side effect of aspirin is heart burn, this can occur at low doses.
– Aspirin was once used as an anti-inflammatory drug as well as a pain killer. This results in the medication working as a block pain. For example, when a person has a headache, it is often the result in restriction in the blood vessels of the brain. Aspirin reduces the swelling and also thins down the blood to help with blood flow through the vessels; this help in reducing pain and prevention of recurring when the painkiller properties of aspirin wear off. Aspirin is also effective in reducing fevers. People who have a history of heart problems and those at risk of heart attacks are often advised by doctors to take daily doses of aspirin as a preventative measure.
The blood thinning properties of aspirin can save a life in the event of a heart emergency by helping some blood to pass through the body until a clot or blockage can be treated. Aspirin is also a known anti-coagulant, which means that it helps to keep the blood thin and prevents clotting. Anti-coagulant is used to prevent blood clots forming within the blood vessels; this can cause heart attack, stroke or other circulatory system problems.

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