Loading...
Thesis
Browse
103 results
Search Results
ThesisItem Open Access PHARMACOLOGICAL EVALUATION OF HERBAL METHIONINE IN METHIONINE DEFICIENCY AND IRON INDUCED STRESS IN BROILERS(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2009-12) SAI GOPAL, T; USHA RANI, M(MAJOR); GOPALA REDDY, A; ANAND KUMAR, AABSTRACT: A total of 120 sexed male broiler chicks of Vencobb strain of day-old age were randomly divided into 8 groups consisting of fifteen chicks in each group. Group 1 was maintained on methionine deficient diet and groups 3, 5 and 7 were supplemented with herbal methionine at level 1 and 2, and synthetic methionine, respectively. Group 2 was maintained as iron added methionine deficient diet and groups 4, 6 and 8 were supplemented with herbal methionine at level 1 and 2, and synthetic methionine, respectively. All the groups were maintained on iso-nitrogenous and iso-caloric diet for a period of 6 weeks. The performance parameters were recorded at weekly intervals. Antioxidant defense profile, biomarkers of hepatic damage, renal damage, protein profile and lipid profile were carried out at 2"d, 4'h and 6th week. At 5m week phytohaemagglutinin (PHA) index and at the end of 6th week histopathological studies were carried out. The methionine deficient and iron added methionine deficient diet groups had a significant (Pe0.05) reduction in body weight, GSH, activity of SOD and catalase, and PHA index, while FCR, and the concentration of TBARS, protein carbonyls and serum creatinine, and the activity of AST were significantly (Pc0.05) increased. Supplementation with herbal methionine at level 1 and 2 respectively in groups 3 and 5 resulted in a marked improvement in all the above parameters as compared to those of methionine deficient diet. Supplementation of herbal methionine at level 2 revealed the performance comparable with synthetic methionine supplementation. Histological abnormalities were also recorded in the liver, kidney, spleen and bursa in all groups, while the groups, 5 and 7 did not reveal any abnormalities on histopathology, while the treated groups 3, 5 and 7 revealed lesions of mild intensity or signs of regeneration. Thus, it is concluded that deficiency of methionine alone, and iron also induces biological damage by means of oxidative stress and the herbal methionine in test offered better performance. The beneficial effects of herbal methionine may be attributed to its antioxidant, anti-stress, hepato-protective principles and biological utilization was as good as synthetic methionine.ThesisItem Open Access MOLECULAR DETECTION AND CHARACTERIZATION OF CLOSTRIDIUM PERFRINGENS TOXIN GENES RESPONSIBLE FOR NECROTIC ENTERITIS IN POULTRY OF ANDHRA PRADESH(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2008-04) PRAVEEN KUMAR, N; VINOD KUMAR, N(MAJOR); RANI PRAMEELA, D; SUJATHA, KABSTRACT: The present study was carried out to investigate the prevalence of Clostridium perfringens toxin genes responsible for necrotic enteritis in poultry of Andhra Pradesh. A total of 691 faecal samples (396 necrotic enteritis suspected and 295 apparently healthy) comprising of cloacal swabs from live birds and intestinal scrapings from dead birds were collected from different districts of Andhra Pradesh viz., Chittoor, Guntur, Nellore, Krishna, East Godavari and West Godavari. Gross pathological studies of affected birds revealed necrosis of the small intestinal mucosa and submucosa with fibrin deposition resulting in pseudo membrane formation and turkish towel appearance was noticed in the small intestine. Microscopically lumen of intestine with fibrinonecrotic material which forms a visible pseudo membrane composed of cell debris, necrotic/distorted villi, inflammatory cells and clumps of bacteria were observed. The samples were inoculated in to fluid thyoglycollate broth and incubated overnight. DNA extracted from 24 h broth cultures by boiling method were screened by multiplex PCR for simultaneous detection of α, β and β2 toxin genes. Out of 396 (broiler 282 & layer 114) necrotic enteritis suspected samples 337/396 (85.1%) were positive for α toxin gene of which 189/337 (56.08%) were β2 toxin gene positive. Out of 295 (broiler 182 & layer 113) apparently healthy samples 61/295 (20.67%) were positive for α of which 4/61 (6.55%) were β2 positive. None of the sample showed amplification of β toxin gene indicating the absence of C. perfringens type C. As some recent studies focused the involvement of NetB toxin in pathogenesis, therefore, uniplex PCR amplification of NetB gene was done from alpha toxin gene positive samples (C. perfringens type A) yielded no positives for NetB toxin gene. From chi square analysis a significant difference (p<0.01) in the prevalence of toxin genes (cpa & cpb2) between necrotic enteritis suspected and apparently healthy at 99% level of confidence with an increased number of positives from necrotic enteritis suspected group. The present research indicates C. perfringens type A along with β2 toxin gene might be responsible for causing necrotic enteritis in broilers and layers in Andhra Pradesh. The multiple sequence analysis of the amplified partial cpa and cpb2 gene sequences revealed 100% homology between the present isolates, and with selected published sequences from GenBank were found to be 98-99% and 94-99% homology respectively. The phylogenetic analysis of the cpa gene of the present C. perfringens isolate (MG600591) with the selected published sequences of cpa revealed the close segregation in distinct clade with cpa gene of broiler isolate of C. perfringens (GU581186) from Iran. The phylogenetic analysis of three cpb2 sequences of present isolates (MF471364; MF471366; KX001813) segregated into close group of poultry originated sequences with exception of MF471365 which segregated in distinct clade with noporcine originated C. perfringens sequence (AY609173) from USA. Since alpha toxin gene (cpa) is considered as signature toxin gene for C. perfringens, amplification of cpa by PCR is considered as confirmative diagnosis of C. perfringens. Hence, in the present study all the PCR positives for cpa (n=398) were isolated by culturing revealed only 221/398 (55.52%) isolates indicating PCR is more sensitive in detecting C. perfringens when compared with isolation by culturing. In the present study culture supernatant of B. subtilis isolated from healthy intestinal contents of birds successfully inhibited C. perfringens by disk diffusion method indicating its importance as a probiotic in controlling necrotic enteritis in poultry.ThesisItem Open Access PATHOLOGICAL CHANGES IN ARSENIC INDUCED TOXICITY AND ITS AMELIORATION IN BROILER CHICKEN(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2009-12) TANJU, SAGI; MADHURI, D(MAJOR); ANJANEYULU, Y; DHANA LAKSHMI, KABSTRACT : The objective of the present work was to evaluate the toxic effects of sodium arsenite, As(III) and its amelioration with Moringa leaf powder @ 0.1% and ascorbic acid @ 200 ppm in broiler chicken. In the present study, chicks of day-old age belonging to Vencobb strain were assigned to six groups (I,II,III,IV,V,VI) consisting of eighteen chicks in each group. Group I served as control (basal diet) and birds in group II were given sodium arsenite @ 200 ppm in basal diet for 42 days. Group III and IV were given Moringa oliefera leaf powder and ascorbic acid @ 0.1% and 200 ppm in basal diet respectively. Group V was fed, diet containing sodium arsenite @ 200 ppm and Moringa leaf powder @ 0.1% for 42 days. Birds in group VI were given with sodium arsenite @ 200 ppm and ascorbic acid @ 200 ppm in basal diet for 6 weeks. Clinically, reduced feed intake and decreased body weight gain were observed in birds fed with sodium arsenite and improvement in these were noted in ameliorative groups. Six birds from each group were sacrificed at fortnightly intervals. Blood, serum and tissue samples were collected. Over all mean values of Hb,PCV and TEC levels in control groups were in normal range where as levels were significantly (P<0.05) decreased in toxin group. The serum biochemical assays showed significant (P<0.05) reduction in total protein, albumin, globulin, A/G ratio while significant (P<0.05) increase in AST, creatinine, GGT was observed in group II birds. Oxidative stress parameters revealed significant (P<0.05) decrease of liver GSH in group II. The ameliorative groups V and VI showed marked improvement in all the above parameters as compared to toxic group. Grossly group II birds revealed enlarged liver with hyperemia, haemorrhages on kidney and liver, reduction in the size of spleen and bursa of Fabricius. Histopathological lesions in group II comprised of severe sinusoidal and venous congestion with mononuclear cell infiltration. Kidney sections showed severe congestion, intertubular haemorrhages, hydropic degeneration, cystic spaces and necrosis of epithelium. Severe depletion of germinal centres was observed in sections of spleen. Depletion of lymphocytes and cystic spaces was observed in sections of bursa of Fabricius. Heart sections revealed complete disruption of muscle fibres and heavy mononuclear cell infiltration. Similar types of gross and histopathological lesions were noted in ameliorative groups of less intensity. In conclusion, sodium arsenite caused a significant toxicity in broiler chicken and its amelioration with Moringa oleifera leaf powder and ascorbic acid was effective in combating the arsenic induced toxicity.ThesisItem Open Access EFFECT OF SUPPLEMENTING RUMEN BY PASS FAT AS CONCENTRATED ENERGY SOURCE ON NUTRIENT UTILIZATION IN MURRAH BUFFALOES(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2009) SRAVAN KUMAR, RAPOLU; RAMANA REDDY, Y(MAJOR); SRINIVASA RAO, D; RAVINDER REDDY, VABSTRACT : With a view to investigate the optimum level of inclusion of calcium soap as concentrate source of energy in the straw based diets of Murrah buffaloes, four iso-nitrogenous rations (12.6% CP) supplemented with 0, 5, 7.5 and 10 per cent calcium soap of red palm oil were formulated and evaluated by in vitro and in sacco techniques. Data from in vitro and in sacco studies revealed that supplementation of calcium soap up to 10% had no adverse effect on DM digestibility/degradability of ration. The rations with 0 to 10 per cent of calcium soap were further evaluated in vivo using four Murrah buffaloes with an average body weight of 221.3±3.60 kg. A metabolic trial was conducted to study the effect of inclusion of calcium soap of red palm oil on nutrient utilization. Dry matter intake (kg/d or g/kg w0.75) in buffaloes was not significantly (P>0.05) affected by the level of protected fat in the diet. There was no significant (P>0.05) difference in the digestibility of proximate principle and cell-wall constituents except EE. The EE digestibility was higher by 15.20, 16.89 and 17.22, percentage units, respectively in buffaloes fed rations with 5, 7.5 and 10 per cent protected fat compared to those fed ration with out protected fat supplementation. All the buffaloes were on positive nitrogen balance. Nitrogen retention expressed as g/d, per cent intake or per cent absorbed in buffaloes was not significantly (P>0.05) different among the rations supplemented with 0, 5, 7.5 and 10 per cent level of protected fat. The DCP values of rations with varying levels of calcium soap of red palm oil were not different from each other. The TDN value of the rations with 5, 7.5 and 10 per cent calcium soap was significantly (P<0.01) higher by 3.18, 4.84 and 6.80 percentage units, respectively as compared to ration with out calcium soap. Further, the DCP and TDN intakes were non-significantly (P>0.05) higher in buffaloes fed rations with 5, 7.5 and 10 per cent level of protected fat than those fed ration with out protected fat. It is concluded from the present study that calcium soap prepared from red palm oil can be used as an energy supplement up to 10% level for Murrah buffalo animals with out affecting DMI and nutrient utilization for improved production.ThesisItem Open Access PHARMACOLOGICAL EVALUATION OF HERBAL NEONATAL CHICK CARE IN BROILERS(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2009-11) CHANDRAVATHY, JADA; GOPALA REDDY, A(MAJOR); USHA RANI, M; ANAND KUMAR, AABSTRACT : A total of 130 sexed male broiler chicks, immediately after hatch, belonging to Vencobb strain were randomly divided into six groups consisting of twenty five chicks each in groups 1, 2, 3 and 4, and fifteen each in groups 5 and 6. Group 1 was maintained on basal diet, group 2 on herbal neonatal chick care @ 6g/chick/day for 2 days after hatching and later continued with basal diet up to 42nd day (6 wks). Group 3 was kept on herbal neonatal chick care @ 8g/chick/day for 2 days after hatching and later continued with basal diet up to 42nd day (6 wks). Group 4 was given FeSO4 @ 0.5% of feed for 42 days (6 wks). Group 5 was given herbal neonatal chick care @ 6g/chick/day for 2 days after hatching and later continued with the FeSO4 @ 0.5% of feed up to 42nd day (6 wks). Group 6 was given herbal neonatal chick care @ 8g/chick/day for 2 days after hatching and later continued with the FeSO4 @ 0.5% of feed up to 42nd day (6 wks). The performance parameters were recorded at weekly intervals. Antioxidant defense profile, biomarkers of hepatic damage, renal damage, lipid profile, protein profile and HI titre in serum were estimated at the end of 4th and 6th wk. Histopathology and estimation of TBARS, GSH, protein carbonyls, HI and phytohaemagglutinin (PHA) index were done at the end of 6th wk. The ferrous sulphate treatment in group 4 resulted in significant (P<0.05) reduction in body weights, protein profile, GSH (6th week), HDL cholesterol, HI titre and PHA index (6th week), while FCR, total cholesterol, LDL cholesterol, triglycerides, TBARS (6th week), ALT, CPK and creatinine were significantly (P<0.05) increased at the end of 4th week and a similar trend was continued at the end of 6th week. Treatment with herbal neonatal chick care in groups 2, 3, 5 and 6 resulted in a marked improvement in all the above parameters as compared to those of ferrous sulphate toxic control group 4 at the end of 6th week. Histological abnormalities were also recorded in the liver, kidney and other tissues in ferrous sulphate toxic control group 4. Groups 1, 2 and 3 did not reveal any abnormalities on histopathology, while the treated groups revealed lesions of mild intensity or signs of regeneration. Thus, it is concluded that ferrous sulphate induces biological damage by means of oxidative stress and the herbal neonatal chick care offered protection and proved beneficial in resisting the adverse effects of stressorThesisItem Open Access EFFECT OF EXOGENOUS ENZYMES IN HIGH FIBRE DIETS AND THEIR IMPACT ON BROILER PERFORMANCE(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2009-10) RAMBABU, D; RAVINDER REDDY, V(MAJOR); QUDRATULLAH, S; REDDY, V.R; KONDAL REDDY, KABSTRACT: Use of unconventional agro-industrial byproducts in poultry diets is restricted due to high level of crude fibre or presence of some deleterious fixtors. The principal components of dietary fibre are the structural carbohydrates which include cellulose, hemicellulose, P-glucans, arabinoxylans and pectins. Inadequate or non-availability of certain enzymes in the digestive tract of chicken reduces their nutrient utilization. A feeding trial of 6 weeks duration was conducted on 240 day-old commercial male broiler chicks procured and allotted randomly into eight treatments. Each treatment was divided into six replicates, with five chicks per replicate and raised in electrically heated battery brooders to evaluate tbe effect of exogenous enzymes on performance of broilers, fed with high and low fibre diets. Eight test diets were prepared by supplementing enzyme preparations to the reference diets i.e., TI, low fibre and T2 high fibre diets respectively. T3, T4 were low and high fibre diets supplemented with fibre degndng enzymes. Similarly, T5, T6 are low and high fibre diets supplemented with fibre degrading enzymes along with protease enzyme. TI. T6 arc low and high fibre diets supplemented with protease enzyme. Feed and water were provided ad libitum throughout the experimental period. The performance of broilers in terms of weight gain, feed intake, feed efficiency, livability, dressing percentage, visceral organs, abdominal fat, provenmculus weights and small intestinal lengths along with immune response to NDV and SRBC were studied. High fib diets (T4, T6, T6) supplemented with enzymes could record significantly l;p-.OS) improved body weight gains during finisher (5-6 week) and overall periods (0-6 weeks) with respect to the'i control diet (T2). Supplementation of protease enzyme to high fibre diet (Ts) resulted in significantly (P4.05) improved body weight gain during starter, finisher and overall periods compared to their correspoding control diet (G). The faad intake and feed-to-gain values were not affected by supplementation of exogenous enzymes either to high or low fibre diets. The livability was also unaffected on enzyme supplementation to both high and low fibre diets. The moisture content of litter values also were not affected by enzyme supplementation. Supplementation of enzymes to high and low fibre diets wuld recorded signific,antly (P4.05) improved dressing percentage, while the organ weights like of proventriculus, giblets, and abdominal fat and small intestine length were dected on enzyme supplementation. The lymphoid organ weights (thymus, spleen and bursa) expressed as percent on live body weight of bird were not affected on supplementation of enzymes either to high or low fibre diets. Humoral immune response to NDV in tenns of increased antibody production was found to be significant (Pc0.05) at 42d age on enzyme supplementation to high fibre diets. Antibody production on low fibre diets with enzyme supplementation was also not influenced. Humoral immune response to sheep RBC, on the antibody production with enzymes supplementation to either high or low fibre diets did not reveal any significant (Ps0.05) effect. However, antibody production was significantly (Pa.05) different among high or low fibre diets and enzyme supplemented diets. Economic calculation revealed that the returns over feed cost on high fibre enzyme supplemented diets were better over the unsupplemented diets and also low fibre enzyme supplemented diets. Supplementation of fibre degrading enzymes (cellulase-420 IU / kg, xylanasA025 IU / kg and pectinase-53 11J / kg) and proteolytic enzyme (protease-5000 U / kg) to high fibre diets wuld result in better weight gains of broilers.ThesisItem Open Access DETECTION OF YERSINIA ENTEROCOLITICA IN LIVESTOCK PRODUCTS BY PCR TECHNIQUE(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2009-09) SHASHIKALA, SANKARAMADDI; KRISHNAIAH, N(MAJOR); MADHAVA RAO, T; GOPALA REDDY, AABSTRACT : The present study was undertaken to standardize PCR assay for detection of Yersinia enterocolitica and Yersinia heat stable enterotoxin from livestock products and compare its efficacy with conventional cultural methods. Primers derived from ail gene and yst B genes used which gave specific amplification products of 425 and 146bp for Y. enterocolitica and yersinia heat stable enterotoxin (yst B) respectively. To determine their suitability to PCR, three different template preparation methods viz. heat lysis, lysis buffer-1and 2 were compared, of which heat lysis was found to be efficient and convenient. The specificity for Y. enterocolitica and yersinia heat stable enterotoxin were tested using primers from ail and yst B genes with Y. enterocolitica and seven other non-Y. enterocolitica strains, which gave specific products at 425 and 146bp for Y. enterocolitica and Yersinia heat stable enterotoxin respecitvely. The minimum detection level with pure Y. enterocolitica culture was 2.0 cfu. Evaluation of two non-selective broths (Tryptic Soya broth and Brain heart infusion broth) and two selective broths (Yersinia selective enrichment broth and Irgasan-ticarcillin-potassium chloride broth) for PCR compatibility, it revealed that ITC was superior followed by YSE, BHI and TSB. Spiking studies were carried out by inoculating with pure culture for Y. enterocolitica in selective enrichment broths (24hr), which revealed that earliest detection time by PCR of Y. enterocolitica was 24 hr and ITC was the most suited selective broth for PCR assay. Screening 300 naturally contaminated samples (25 each of pork, pork tonsils, pork tongues, pork surface swabs, swine faeces, swine farm water, milk, beef, cattle faeces, dairy farm water, chicken and poultry farm water samples) revealed 132 (14, 23, 23, 10, 9, 21, 4, 5, 4, 8, 6 and 5 respectively for the above samples) positive for Y. enterocolitica, out of which 56 samples (5,10,9,4,2,9,2,2,1,5,4 and 3 respectively) were positive for yst B gene.ThesisItem Open Access TOXICOLOGICAL STUDIES IN THE PROGENY OF DAMS TREATED WITH METHIMAZOLE, MONOCROTOPHOS AND LEAD ACETATE(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2009-12) VANITHA SREE, K; GOPALA REDDY, A(MAJOR); USHA RANI, M; ANJANEYULU, YABSTRACT : The present study was aimed to evaluate the toxicological effects in the progeny of dams treated with methimazole, monocrotophos and lead acetate. Albino rats of Wistar Kyoto strain were divided into 5 groups and treated as follows: Group 1 served as sham control, 2 received methimazole, 3 received monocrotophos, 4 received lead acetate and 5 received combination of monocrotophos and lead acetate. Thyroid hormone profile was recorded on 14th day of gestation in dams. Average body weights were recorded at monthly intervals in pups born to test dams. On 21st day and 90th day, TBARS, protein carbonyls and GSH of liver and heart, histopathology of liver, heart and kidney were studied. Activity of Na+ K+ ATPase and Mg2+ATPase of liver and heart, and histopathology of brain and testis were studied on 90th day. T3, T4, RBC, Hb, HDL cholesterol, serum calcium, GSH, Na+ K+ ATPase and Mg2+ATPase were significantly (P < 0.05) decreased in all the test groups, while TSH, TBARS, protein carbonyls, blood glucose, triglycerides, serum LDH, intra-testicular LDH, total WBC count and serum troponins were significantly (P < 0.05) increased. Group 1 did not reveal any abnormalities on histopathology. Groups 5 (MCP + lead acetate) and 2 (methimazole) showed marked congestion and degenerative changes in heart, kidney, brain and testis, while thyroid gland showed moderate disruption of follicular epithelium. Groups 3 and 4 revealed mild abnormalities on histopathology. From this study, it is concluded that both monocrotophos and lead acetate might have a possible influence on thyroid gland of dams as the thyroid hormone levels, which play an important role in early peri-natal life in the proper development of foetus, were significantly altered and were comparable among the treatments in dams and the histological sections of thyroid obtained from progeny of MCP + lead acetate treated rats showed lesions comparable to those seen with methimazole.ThesisItem Open Access DETECTION OF VIBRIO PARAHAEMOLYTICUS IN FOODS BY PCR TECHNIQUE(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2009-09) SUBHASHINI, NELAPATI; KRISHNAIAH, N(MAJOR); MADHAVA RAO, T; ANJANEYULU, YABSTRACT : The present study was undertaken to standardize PCR assay for detection of Vibrio parahaemolyticus and their toxins from foods and compare its efficacy with conventional cultural methods. Primers derived from toxR gene and tdh and trh genes were used which gave specific amplification products of 368, 623 and 460 bp for V. parahaemolyticus and toxins (tdh and trh) respectively. To determine their suitability to PCR, four different template preparation methods viz. genomic DNA extraction, heat lysis, lysis buffers-1 and 2 were compared, of which heat lysis was found to be efficient and convenient. The specificity for V. parahaemolyticus and toxins were tested using primers from toxR, tdh and trh genes with V. parahaemolyticus and nine other non-V. parahaemolyticus strains, which gave specific products at 368, 623 and 460 bp for V. parahaemolyticus and toxins respectively. The minimum detection level was 2.5 cfu for both V. parahaemolyticus and toxins. Evaluation of two non-selective broths, i.e Brain Heart Infusion Broth (BHI) and Tryptic Soy Broth (TSB) and two selective broths, i.e Alkaline Peptone Water Broth (APW) and Salt Polymixin Broth (SPB) for PCR compatibility, it was revealed that SPB was superior followed by APW, BHI and TSB. Spiking studies were carried out by inoculating with pure culture of V. parahaemolyticus in selective enrichments broths (8h and 18h), which revealed that earliest detection time for V. parahaemolyticus by PCR was 18h and SPB was the most suited selective broth for PCR assay. Screening of 280 naturally contaminated samples (35 each of fresh water fish, sea fish, fresh water prawn, marine shrimp, crab, fresh water, marine water and mutton samples) revealed 165 positive for V. parahaemolyticus (fresh water fish-17, sea fish-20, fresh water prawn-24, marine shrimp-33, crab-27, fresh water-14, marine water-28 and mutton-2) out of which 24 carried only tdh (fresh water fish-2, sea fish-4, fresh water prawn-3, marine shrimp-4, crab-4, fresh water-2, marine water-5 and mutton-nil), 95 carried only trh (fresh water fish-12, sea fish-16, fresh water prawn-9, marine shrimp-13, crab-15, fresh water-12, marine water-18 and mutton-nil) and 15 carried both tdh and trh (fresh water fish-1, sea fish-2, fresh water prawn-1, marine shrimp-3, crab-2, fresh water-2, marine water-4 and mutton-nil) simultaneously.