Physiological responses of dietary tryptophan fed Labeo rohita to temperature and salinity stress

Two experiments were conducted to elucidate the possible effects of dietary L‐tryptophan (TRP) in Labeo rohita based on growth performance and physio‐biochemical responses. In the experiment I, a 60‐day feeding trial was carried out to elucidate the effects of dietary TRP enrichment on growth performance and physio‐biochemical responses. In the experiment II, the TRP pre‐fed L. rohita, from experiment I, was exposed to temperature and salinity stress to evaluate stress‐mitigating efficacy of TRP. In L. rohita, dietary supplementation of TRP showed significant effect on weight gain percentage and feed conversion ratio but not on blood glucose. A significant increase in RNA content and RNA/DNA ratio upon TRP supplementation was observed and was positively correlated with growth performance. The results of experiment II indicated that weight gain percentage, serum T3 and T4 levels were significantly reduced in groups that were exposed to temperature and salinity stress and fed diets without TRP supplementation. However, dietary supplementation of TRP significantly augmented weight gain percentage in stress‐exposed groups. Tryptophan supplementation helped in bringing back T3 and T4 levels comparable with control. A significant increase in superoxide dismutase, catalase, Adenosine triphosphatase, blood glucose and serum cortisol was observed in temperature‐ and salinity‐exposed groups fed without TRP‐supplemented diets. However, TRP supplementation was found to be effective in restoring the above parameters. The results of these experiments suggest that dietary TRP supplementation augments growth, lowers energy demand and helps in mitigating thermal and salinity stress in L. rohita.     

Immunosuppressive effects of aflatoxin B1 in Indian major carp (Labeo rohita).

Aflatoxin B1 (AFB1) is a potent immunomodulator in endotherms. The experiment was carried out to study the immunosuppressive nature of AFB1 in one ectothermic species of Indian major carp. Graded levels (0, 1.25, 5.00 mg/kg of body weight) of purified AFB1 were intraperitoneally (i.p.) injected into rohu (Labeo rohita) fingerlings weighing 30-50 g, and the fish were observed for a period of 90 days. At the end of the trial, blood samples were collected from the control group as well as the AFB1 injected fish and were screened for nitroblue tetrazolium (NBT) assay, serum total protein, albumin, globulin, albumin-globulin ratio (A:G), serum bactericidal activity and bacterial agglutination titre against Edwardsiella tarda. The aflatoxin-treated fish revealed a reduction of total protein, globulin levels, bacterial agglutination titre, NBT and serum bactericidal activities, as well as an enhanced A:G ratio without change in albumin concentration, irrespective of dose levels of toxin treatment, when compared to the control group. Thus, AFB1 proved to be immunosuppressive in rohu even at the lowest dose (1.25 mg/kg body weight) of toxin treatment. This could be of economic significance in intensive culture systems of rohu.     

Ultrastructural Changes in the Adrenal Cortex of Rats Exposed to Extremely Low Temperatures

The response of the adrenal cortex to low temperature in relation to the length of the exposures is studied. The morphological changes of the adrenal cells, mainly in the Zona fasciculata, are examined. Thirty white rats of both sexes, divided into six groups, were exposed during time intervals of different lengths to temperatures ranging between – 12° and – 15°C. The cellular modifications encountered by means of the electron microscope at the level of the mitochondria, the smooth endoplasmic reticulum and the liposomes are analyzed and compared with the functional situation of the gland in an effort to establish the existence of a correlation between the morphology and the function.     

Gelatinized to non-gelatinized starch ratio in the diet of Labeo rohita: effect on digestive and metabolic response and on growth

A 60-days experiment was conducted to study the optimum gelatinized (G) to non-gelatinized (NG) starch ratio in the diet of Labeo rohita juveniles with respect to digestive and metabolic response and on growth. Two-hundred and thirty-four juveniles (avg. wt 2.53 ± 0.04 g) were randomly distributed in six treatment groups with each of three replicates. Six semipurified diets either containing NG and/or G corn starch viz., T₁ (100% NG, 0% G starch), T₂ (80% NG, 20% G starch), T₃ (60% NG, 40% G starch), T₄ (40% NG, 60% G starch), T₅ (20% NG, 80% G starch) and T₆ (0% NG, 100% G starch) were prepared. The dry matter digestibility and carbohydrate digestibility were highest (p < 0.05) in T₆ group and lowest in T₃ and T₄ groups. The amylase activity in intestine increased as G:NG level increased in the diet. Protease activity in intestine was highest in T₆ group and lowest in T₁ group. Similar trend was recorded for specific growth rate, protein efficiency ratio and apparent net protein utilization. Liver glycogen, hepatosomatic index and blood glucose level increased linearly with the increasing level of G starch in the experimental diet. The results indicate that higher nutrient digestibility and growth was recorded either at low (20% G starch, T₂) or high (100% G starch, T₆) G starch fed group. But high G starch fed group (T₆) exhibits higher liver glycogen and blood glucose level, which may lead to stress due to long-term feeding. Hence, it is suggested that 20% G and 80% NG starch is optimum for better nutrient digestibility and growth in L. rohita juveniles.     

Effects of dietary pyridoxine on haemato-immunological responses of Labeo rohita fingerlings reared at higher water temperature

A sixty-day feeding trial was carried out to elucidate the effect of dietary pyridoxine (PN) on hemato-immunological parameters in Labeo rohita fingerlings exposed to an elevated temperature (ET) of 33 °C. Two hundred and seventy fingerlings (6.71 ± 0.32 g) were randomly distributed into six treatments in triplicates (15 fish/tank). Five iso-nitrogenous (356.3 ± 2.7 g/kg crude protein) purified diets were prepared with graded levels (0, 10, 50, 100 and 200 mg/kg diet) of PN. Six treatment groups were P10T26 (10 mg PN + ambient temperature (26 °C), P0T33 (0 mg PN + ET), P10T33 (10 mg PN + ET), P50T33 (50 mg PN + ET), P100T33 (100 mg PN + ET) and P200T33 (200 mg PN + ET). The responses of L. rohita fingerlings to dietary PN were assessed in terms of growth and by analysing some hemato-immunological parameters. When PN supplementation increased from 10 to 200 mg/kg diet, a significant improvement in weight gain percentage was observed. Erythrocyte (RBC) count, leucocyte (WBC) count, haemoglobin content and respiratory burst activity were significantly (p < 0.05) higher in 100 mg PN/kg diet-fed group than their non-PN-fed counterpart. Also, serum albumin, globulin and lysozyme activity were found significantly (p < 0.05) higher in PN-fed groups. The overall results indicated that dietary PN supplementation at 100 mg/kg diet may reverse the negative effects caused by ET and may protect the haemato-immunological status of L. rohita fingerlings reared at higher water temperature.     

Higher water temperature enhances dietary carbohydrate utilization and growth performance in Labeo rohita (Hamilton) fingerlings

A 60-day experiment was conducted to delineate the effect of three dietary levels of gelatinized carbohydrate (GC) on growth, nutrient-utilization and body composition of Labeo rohita fingerlings (avg. wt 6.5 ± 0.3 g) reared at two water temperatures (ambient-AT (26 ± 0.8 °C) and 32 °C). Two hundred and sixteen fingerlings were randomly distributed into six treatments in triplicates. Three semi-purified isonitrogenous diets were prepared with graded levels of GC viz. D(1) : 40%, D(2) : 50% and D(3) : 58%. Growth rate, feed efficiency and protein efficiency ratio were significantly (p < 0.05) higher in 50% GC and 32 °C reared groups than their AT counterparts. Hepato Somatic Index was higher in AT reared groups compared to 32 °C reared counterparts. Apparent digestibility co-efficient of carbohydrate was significantly (p < 0.05) higher at 32 °C reared groups but decreased with increasing carbohydrate (GC) levels. Fish reared at 32 °C showed significantly (p < 0.05) higher amylase, protease and hexokinase activities while glucose-6-phosphate dehydrogenase and glucose-6-phosphatse were significantly (p < 0.05) higher at ambient temperatures. The results obtained in present study indicated that L. rohita could utilize higher level (50%) of dietary carbohydrate at 32 °C.     

Effect of Dietary Supplementation with Achyranthes aspera Seed on Larval Rohu Labeo rohita Challenged with Aeromonas hydrophila

Abstract

Larval rohu Labeo rohita were fed four different diets: three of the diets contained Achyranthes aspera (prickly chaff-flower) seeds at 0.10% (D1), 0.25% (D2), or 0.50% (D3); the fourth diet was a control diet (D4; no A. aspera supplementation). After 70 d, the rohu were injected intraperitoneally with live Aeromonas hydrophila. Mortality of fish was recorded for 7 d. In the D4 group, the first mortality was observed within 12 h of exposure, whereas in the D1–D3 treatment groups, mortality was first observed at 24 h postexposure. In the D4 group, 50% of fish died within 72 h of exposure, whereas in the D3 group, 10–15% mortality occurred between 72 and 84 h. The cumulative mortality rate was 50% for D4, 40% for D1, 35% for D2, and 15% for D3. Total tissue protein level in the larvae was higher for the D2 and D3 groups than for the other groups. Glutamic oxaloacetic transaminase, glutamate pyruvate transaminase, and thiobarbituric acid reactive substance levels were significantly lower in D3 larvae than in the other groups, whereas lysozyme and nitric oxide synthase levels were significantly higher in D3 larvae compared with the other groups. Dietary supplementation with A. aspera seeds at the 0.50% level provided protection against oxidative stress, prevented tissue damage, and enhanced disease resistance in rohu larvae.

Received December 26, 2011; accepted May 7, 2012     

Immunomodulation by dietary vitamin C in healthy and aflatoxin B1-induced immunocompromised rohu (Labeo rohita)

The aim of this study was to examine the immunomodulatory effect of high levels of dietary vitamin C in healthy and immunocompromised rohu (Labeo rohita) treated with aflatoxin B1 (AFB1). Four groups of rohu were fed experimental diets containing either no vitamin C or supplemented with vitamin C at 500 ppm for 60 days. On the first day of feeding, one group fed the high vitamin C diet and one fed the vitamin C deficient diet, were injected intraperitoneally with a single doses of AFB1 at 1.25 mg kg(-1) body weight. The effect of AFB1 and high dietary vitamin C on specific and non-specific immunity, and disease resistance against Aeromonas hydrophila were examined in the rohu. The ability of vitamin C to counteract immunosuppression induced by AFB1 was also examined. Specific immunity indicated by haemagglutination and haemolysin titres against sheep red blood cells (SRBC), and bacterial agglutination appeared to be unaffected by either the AFB1 treatment or the vitamin C enriched diet. A significant reduction was observed in the non-specific immunity of AFB1-treated fish, however, indicated by lowered bactericidal and lysozyme activities. High dietary vitamin C, on the other hand, enhanced the non-specific immunity of fish, including an enhanced phagocytic ratio and increased serum lysozyme activity. Feeding a high level of dietary vitamin C to AFB1-treated fish increased these parameters to levels similar to those found in control fish. High dietary vitamin C significantly (p < 0.05) enhanced protection against Aeromonas hydrophila infection in both healthy and immunocompromised fish. Results from this study help to establish the beneficial effect of dietary vitamin C on AFB1-induced immunosuppression, as well as confirming the immunostimulatory effect of vitamin C in rohu.     

Structural and Ultrastructural Differentiation of Cell Types in the Gills of the Tench (Tinca tinca, L.)

This study used 22 tench of 25-30 cm in length and weighing roughly 200 g. After sacrifice, the gill arch was extracted and fixed in 5% glutaraldehyde for structural, ultrastructural and morphometrical study. Tench gills consist of primary lamellae which are perpendicular to numerous secondary lamellae in which the respiratory barrier is located. A study was made of the structures which make up that barrier and of the multi-layered epithelium situated in the interlamellar space, which is closely linked to maintaining of internal balance in the fish.     

Effect of alternate day feeding strategy of sub-optimal protein level on haemato-biochemical responses in Labeo rohita (Hamilton) juveniles

A feeding trial was conducted for 60 days to delineate the effect of alternate day feeding strategy of sub‐optimal protein level on haematological parameters, serum parameters and phagocyte respiratory burst activity (NBT) in Labeo rohita juveniles. One hundred and thirty‐five fingerlings (1.87 ± 0.01–2.26 ± 0.05 g) were distributed in triplicate groups of each treatment, and fish were fed at 5% body weight daily. Three experimental isocaloric (401.32–410.28 kcal/100 g) diets of 30%, 25%, and 20% crude protein designated as diet A, diet B, and diet C respectively, were prepared, using locally available feed ingredients. Three different feeding schedules of normal protein diet continuously (diet A‐30%), alternate feeding of 1‐day diet A followed by 1‐day diet B (1A/1B) and alternate feeding of 1‐day diet A followed by 1‐day diet C (1A/1C) were tested. The total erythrocyte count and haemoglobin content was significantly (p < 0.05) enhanced in the group T1 fed (1A/1B), and the lowest count was recorded in the group T2 fed (1A/1C). Total leucocyte counts, total serum protein, and serum globulin were higher in the group T1 fed (1A/1B) and lower in the group T2 fed (1A/1C) as compare to control. The respiratory burst activity (NBT) of blood phagocytes and serum A‐to‐G ratio was recorded significantly difference in among the treatment groups. Based on the results of the present study, it is concluded that alternate feeding of 1‐day diet A followed by 1‐day diet B (1A/1B) is equally effective and promote the immunity in Labeo rohita juveniles.     

Expression of immune-related genes in rohu Labeo rohita (Hamilton) by experimental freshwater lice Argulus siamensis (Wilson) infection

The crustacean ectoparasite, Argulus poses one of the major threats to carp culture due to absence of any suitable control measure. The study was undertaken to determine the expression of immune-related genes in three major immunocompetent organs viz., kidney, skin and liver of rohu (Labeo rohita) during experimental freshwater lice Argulus siamensis infection. Results showed that the expression of TLR 22-like, lysozyme G and β2-microglobulin genes in kidney was significantly (P ≤ 0.05) down-regulated in lice-infected fish. On the other hand, no significant difference (P>0.05) in CXCa, lysozyme C, TNFα and complement component 3 (C3) expression was found between uninfected control and different degrees of lice infected fish. In the skin, the expression of TLR 22-like and TNFα genes were significantly up-regulated whereas that of C3 was significantly (P ≤ 0.05) down-regulated in lice-infected fish with respect to control fish. The expression of CXCa, lysozyme C and transferrin was not detected in the skin samples of fish. In the liver, the expression of CXCa, lysozyme G, β2-microglobulin and transferrin was significantly (P≤0.05) up-regulated in lice-infected fish with respect to control fish whereas expression of C3 was significantly (P ≤ 0.05) down-regulated in lice-infected fish. The expression of TLR 22-like, lysozyme C, TNFα genes was not detected in the liver samples of fish. This study indicates that majority of the genes showed down-regulation in kidney tissue whereas up-regulation in liver and skin tissues except C3 in Argulus-infected fish. We show that infection with this parasite irrespective of intensity can also result in immune gene expression changes in tissues situated away from the site of parasite attachment and feeding. The information obtained here could be useful towards understanding the susceptibility of rohu to argulosis and mechanisms involved in protection of rohu to ectoparasitic infections, which is causing immense economic losses to freshwater aquaculture sector.     

Haemato-immunological and stress responses of Labeo rohita (Hamilton) fingerlings: effect of rearing temperature and dietary gelatinized carbohydrate

A feeding trial of 70‐days was carried out to study the haemato‐immunological and stress responses of Labeo rohita fingerlings reared at two water temperatures [ambient (Amb) – 27 °C and 32 °C] fed with graded levels of gelatinized corn carbohydrate (GC). Two hundred and sixteen fingerlings were randomly distributed into six treatment groups in triplicate. Three semi‐purified diets were prepared containing 30% crude protein with graded levels of GC 40%, 50% and 58%. The six treatment groups were T₁ (40% GC × Amb), T₂ (40% GC × 32 °C), T₃ (50% GC × Amb), T₄ (50% GC × 32 °C), T₅ (58% GC × Amb) and T₆ (58% GC × 32 °C). The blood glucose level was significantly (p < 0.05) lowered in groups fed with 58% GC level. Neither dietary GC levels nor temperature had a significant (p > 0.05) effect on serum cortisol and superoxide dismutase activity. Lysozyme activity was significantly higher (p < 0.05) in T₁ during pre‐ and post‐challenge period while temperature alone had a significant (p < 0.05) effect on post‐challenge Nitroblue Tetrazolium and found higher at 32 °C. A significant effect of GC levels and rearing temperature was recorded on WBC in the pre‐ and post‐challenge period. Highest pre‐challenge WBC was observed in T₄ group and in the post‐challenge period T₁ group recorded maximum. Water temperature had significant effect on pre‐challenge haemoglobin content, highest being at 32 °C (T₂). A significant (p < 0.05) effect of rearing temperature and dietary GC level on total serum protein and albumin was also observed. Highest total serum protein and albumin was recorded in T₁ and globulin in T₂. Percentage survival after challenging with Aeromonas hydrophila was highest in T₁ followed by T₃ group and lowest in T₆. The results obtained in the present study suggest that L. rohita fingerlings may utilize higher levels of dietary GC at higher temperature (32 °C) but may affect its immunity status.     

Non‐gelatinized starch influences the deposition of n‐3 fatty acids in the muscle of a tropical freshwater fish,Labeo rohita

A 60‐day feeding trial was conducted to study the influence of gelatinized (G) to non‐gelatinized (NG) starch ratio in the diet on fatty acids profiles and oxidative status in Labeo rohita fingerlings. Two hundred and thirty‐four fingerlings (average weight: 2.53 g) were distributed in six treatment groups with each of three replicates. Six semi‐purified diets either containing NG and/or G corn starch (42.43%) viz., T₁ (100% NG and 0% G starch), T₂ (80% NG and 20% G starch), T₃ (60% NG and 40% G starch), T₄ (40% NG and 60% G starch), T₅ (20% NG and 80% G starch) and T₆ (0% NG and 100% G starch) was fed to respective groups. Catalase, superoxide dismutase and malic enzyme activities decreased linearly with the increasing level of G starch, whereas reverse trend was found for glucose‐6‐phosphate dehydrogenase. Total saturated fatty acids in muscle increased with the increasing level of G starch in the diet. Total n‐3 fatty acids decreased linearly with the increasing level of G starch in the diet. Among the n‐3 fatty acids, linolenic acids content was more in NG starch fed group. Similarly, eicosapentaenoic acid contents gradually decreased with increasing level of G starch content. The n‐6/n‐3 ratio was higher in G starch fed group. This suggests that dietary starch type may be manipulated for quality improvement of fish flesh.     

Serum antibody response of Indian major carp, Labeo rohita to three species of pathogenic bacteria; Aeromonas hydrophila, Edwardsiella tarda and Pseudomonas fluorescens

The immune response to mixed whole cell antigens of Aeromonas hydrophila, Edwardsiella tarda and Pseudomonas fluorescens, the common Gram negative bacterial pathogens associated with diseases of Indian major carps were evaluated for their efficacy in triggering antibody responses in rohu, Labeo rohita (Ham.). The rohu yearlings were either immunized with antigens from single bacterial strain, A. hydrophila, E. tarda and P. fluorescens or a combination of all three. An antibody response was detected at 1st week post immunization that rose significantly (p<0.05) at 4th week post immunization in all the immunized groups. The antibody level started declining after 8th week but persisted up to 10th week post immunization in all the immunized groups. Similarly, no significant difference (p>0.05) in the antibody level was found between groups immunized with single and mixed bacterial antigens. Moreover, the use of mixed bacterial antigens did not jeopardize the specific immune response to the vaccine components. Upon challenge with single pathogen, a high relative percent survival was recorded in the group immunized with mixed bacterial antigens and was comparable to those fish immunized with the single bacteria.     

Aspects on The Ecology of Labeo Capensis and Labeo Umbratus in The Vaal River

Studies on bivalves in the Swartkops estuary have indicated that spatfall occurs during late summer. After adult populations had been dtc'nvttfrrt by floods in 1971 spat made up a large proportion of the bivalve population in 1973. Growth rates vary at different intertidal levels and in different parts of the estuary and growth has been shown to vary seasonally. In two species age determinations from growth rings corresponded well with age determinations from size frequency histograms.     

Respiratory Characteristics of the Blood of Labeo Umbratus (SMITH) and Labeo Capensis (Smith)

Various respiratory characteristics of mudfish blood were investigated. Oxygen and carbon dioxide contents of mixed venous blood were 1,17±0,53 and 11,35±1,65 vol. % respectively. Blood oxygen dissociation curves showed P₅₀ values of 11,84±3,1; 13,77±2,0 and 18,05±4,7 mm Hg at CO₂, concentrations of 0-1%, 4-5% and 9-10%. A marked Bohr effect of —0,59 was observed and the Haldane effect was found to be relatively low. No differences could be observed between the characteristics of L. umbratus and L. capensis blood.     

Ultrastructural findings in horses with chronic obstructive pulmonary disease (COPD). I: Alterations of the larger conducting airways

Extensive light and electron microscope studies of the conducting airways were carried out in 28 horses with varying degrees of clinically manifested chronic obstructive pulmonary disease (COPD) and in 8 horses with normal lungs. The principal ultrastructural changes were found in the ciliated cells. There was focal loss of ciliated cells, which were replaced by undifferentiated cells in a largely hyperplastic epithelium, and some horses, independent of the degree of severity of the disease, showed various types of ciliary malformation. The finding of dilated intercellular clefts and accumulations of mast cells was interpreted as morphological evidence of non-specific mucosal hyperreactivity. Interstitial cells with intracytoplasmal crystal inclusions, the cause of which is not clear, were seen in many horses. Comparison between the clinical diagnosis and the morphological findings showed partial correlation. The ciliary loss, the appearance of peribronchial inflammatory processes and the occurrence of intracytoplasmic crystalline inclusions showed a positive correlation. In 12 of 28 cases, the severity of clinical diagnosis agreed with the degree of the morphological findings. The changes in the conducting airways were interpreted pathogenetically as reactive processes to changes in the small airways in the course of equine COPD.     

Characterization and Inhibition of Nitrite Uptake in Shortnose Sturgeon Fingerlings

Abstract

Efforts are underway to culture the endangered shortnose sturgeon Acipenser brevirostrum for possible reintroduction. As part of a larger project to develop culture techniques for this species, the uptake of nitrite was evaluated in fingerlings (16.5 ± 4.85 g; mean ± SD). Plasma nitrite concentrations increased significantly with exposure time (0–5 d) and dose (0–4 mg nitrite-N/L). Shortnose sturgeon fingerlings were able to concentrate nitrite in their plasma to more than 63 times the environmental concentration. Chloride, as either sodium chloride or calcium chloride, partially inhibited nitrite uptake. However, calcium chloride was a better inhibitor. After previous exposure (2 d at 2.13 ± 0.080 mg nitrite-N/L) plasma nitrite-N decreased from 165.5 to 36.7 mg/L during a 3-d simultaneous exposure to 2.13 ± 0.080 mg nitrite-N/L and treatment with 40 mg chloride/L as calcium chloride. The addition of calcium chloride to the water appeared to be an effective means of preventing nitrite uptake and treating nitrite toxicity in hatchery-reared shortnose sturgeon fingerlings.     

二甲苯对小鼠肺组织损伤的初步研究

为了研究二甲苯对小鼠肺组织的损伤情况,以健康成年小鼠作为试验动物,分3个试验组(低、中、高剂量组)1个对照组,给试验组腹腔注射不同剂量二甲苯(0.125、0.25、0.5 mL/kg),给对照组腹腔注射生理盐水.15 d后剖检取材,采用石蜡切片和HE染色技术研究二甲苯染毒后小鼠肺组织结构的变化特点;采用分光光度法测定小...