Responses of Atlantic cod <Emphasis Type="Italic">Gadus morhua</Emphasis> head kidney leukocytes to phytase produced by gastrointestinal-derived bacteria

This study identified phytase-producing bacteria that were previously isolated from the gastrointestinal tract of Atlantic cod, Gadus morhua and determined its effect on head kidney leukocytes. Out of the 216 bacterial strains tested, the two phytase producers were identified as Pseudomonas sp. and Psychrobacter sp. based on their 16S rDNA sequence. Crude phytase from these two bacterial strains was produced employing the shake flask method. Even though the total protein of the crude phytase was not significantly different for the two bacteria, the phytase activity of the crude enzyme produced by Pseudomonas sp. (97.1 ± 16.7 U) was significantly higher than that of the enzyme from Psychrobacter sp. (75.9 ± 2.4 U). When cod head kidney leukocytes were incubated with the crude phytase (50 μg ml⁻¹), it resulted in enhanced cell proliferation, higher myeloperoxidase, and acid phosphatase activities. Extracellular responses—respiratory burst activity and hydrogen peroxide production were not enhanced by the crude enzyme. As a consequence, the growth of two pathogenic bacteria Aeromonas salmonicida and Vibrio anguillarum was not suppressed by the supernatants obtained from head kidney leukocytes incubated with the crude bacterial phytase. Thus, the enzyme from phytase-producing intestinal bacteria of Atlantic cod can stimulate intracellular head kidney leukocyte activities but not the production of extracellular substances that are involved in antibacterial response. These have implications on the potential use of bacterial phytase as feed supplement to boost cellular immune response of the fish and could be employed as a health management strategy in culture systems.     

Probiotic bacteria <Emphasis Type="Italic">Lactobacillus rhamnosus</Emphasis> influences the blood profile in rainbow trout <Emphasis Type="Italic">Oncorhynchus mykiss</Emphasis> (Walbaum)

This paper reports the effect of feeding probiotic diets on blood profiles in rainbow trout. Two experiments were performed: in the first, fish of average weight 75 g were offered either a commercial feed or the same incorporated with 10⁹ CFU g⁻¹ of lactic acid bacteria Lactobacillus rhamnosus for 30 days; in the second study performed for a similar duration, fish of average weight 126 g were offered formulated diets that either contained the same bacteria in heat-killed or freeze-dried form (nearly 10¹¹ CFU g⁻¹), or the basal diet without the bacteria. Blood samples were collected at different times after commencement of probiotic feeding to determine the total cholesterol, triglyceride contents, the plasma alkaline phosphatase activity, plasma protein and hematocrit value. The plasma cholesterol significantly increased upon probiotic feeding in the first experiment. A significant elevation (P < 0.05) of plasma cholesterol and triglyceride and alkaline phosphatase activity level was found in the freeze-dried probiotic fed groups at 20 and 30 days postfeeding. This was concomitant with the increased plasma protein and hematocrit values in FD group at 20 and 30 days. Likewise, the heat-killed probiotic fed group registered significantly high values of triglycerides, alkaline phosphatase activity, and plasma protein compared to the control diet fed groups after 20 days of feeding. Thus, alterations in the blood profiles could serve as supplementary information when examining the benefits of probiotics for fish.     

Hepatoprotective activity of Calotropis procera flowers against paracetamol-induced hepatic injury in rats

Hydro-ethanolic extract (70%) of Calotropis procera flowers was prepared and tested for its hepatoprotective effect against paracetamol-induced hepatitis in rats. Alteration in the levels of biochemical markers of hepatic damage like SGPT, SGOT, ALP, bilirubin, cholesterol, HDL and tissue GSH were tested in both treated and untreated groups. Paracetamol (2 g/kg) has enhanced the SGPT, SGOT, ALP, bilirubin and cholesterol levels and reduced the serum levels of HDL and tissue level of GSH. Treatment with hydro-ethanolic extract of C. procera flowers (200 mg/kg and 400 mg/kg) has brought back the altered levels of biochemical markers to the near normal levels in the dose dependent manner.     

Phosphorus retention efficiency in rainbow trout fed diets with low fish meal and alternative protein ingredients

ABSTRACT:   Two experiments were conducted to evaluate feed quality and body phosphorus (P) and nitrogen (N) retention by rainbow trout fed test diets with low amounts of fish meal content and alternative low-P protein sources. A fish meal (FM) diet was used as control. Fish weighing 2.0 g and 134.7 g on average were reared with the experimental diets for 30 weeks and 15 weeks, respectively. The experimental diets had a good growth rate and feed utilization. In the first experiment the P retention was higher in the group of fish fed test diets (56 and 69%) compared to the FM-based diet (36%); N retention rates were similar for all groups. In the second experiment, fish were fed the test diet that had the best P retention in the earlier experiment. The P retention was lower than the values obtained in the first experiment (smaller fish), but still higher in the group of fish fed the low FM diet in comparison with the control group (36.0% and 22.2%, respectively). This represents a P loading into the water of 5.9 kg/t and 12.8 kg/t production for the test and the control diet, respectively. Therefore, low-P-loading diets for rainbow trout can be developed through the appropriate combination of alternative protein sources.     

Molecular diagnosis of francisellosis,a systemic granulomatous inflammatory disease in Atlantic cod, <Emphasis Type="Italic">Gadus morhua</Emphasis> L

A PCR-based assay for the detection of Francisella noatunensis causing francisellosis in Atlantic cod, Gadus morhua has been developed. Seven sets of primers targeting the flanking regions of the genes (rpoA, sdhA, atpA, rpoB, pgm, groEL and 16S rRNA) of the pathogen were designed. Among the primers, groEL was found to be the most suitable gene candidate for detecting the pathogen, due to its high sensitivity at various annealing temperatures and specificity in detection. The detection limit of the assay was 100 pg of bacterial DNA per milliliter or 100 fg bacterial DNA (approximately 50 genome equivalents) per PCR reaction, however, the sensitivity of the reaction decreased by 1 log dilution in the presence of 1 mg mL⁻¹ of serum and mucus samples as inhibitors. Nevertheless, the assay can potentially be used as a direct and non-lethal method to detect the pathogen in fish. Thus this PCR assay is a specific and sensitive molecular method to diagnose francisellosis in Atlantic cod, and will be helpful for controlling the infection through prompt detection of the disease in farms.     

In vitro adherence of two candidate probiotics from Atlantic cod and their interference with the adhesion of two pathogenic bacteria

The potential of two candidate probiotic bacteria (GP21 and GP12), isolated from the gut of Atlantic cod, to adhere to primary cultures of the epithelial cells from the different regions of the intestine and to interfere with the adhesion of two pathogens, Vibrio anguillarum and Aeromonas salmonicida subsp. salmonicida were investigated. The intestinal isolates showed clear preference in adhering to the cells from the different intestine segments. GP12 adhered strongly to the fore- and mid intestine cells. The adherence of GP21 was most to the cells from the hind intestine followed by those from the mid-segment. The adhesion of V. anguillarum was affected by both GP21 and GP12; GP12 interfered through competition, but a specific mode of action was not observed for GP21. In the case of A. salmonicida, competition was the principal mechanism by which GP21 interfered with their adhesion, while exclusion mechanism was favoured by GP12. In addition, GP21 was more auto-aggregative than GP12, but the latter was more co-aggregative with both the pathogens. The isolates were also capable of lowering lactate dehydrogenase activity compared to that by the pathogen and they reduced the caspase-3 activity in the epithelial cells from the hind intestine, to which the pathogens adhered the most. Thus it could be concluded that the adhesion of the candidate probiotics is segment-specific and their interference with the adhesion of pathogens is dependent on both source of the epithelial cells and the mechanism adopted by the isolates. This information is novel in the case of fish and the manner in which potential probiotic organisms interfere with the pathogen adhesion provides supportive information for disease control.     

A laboratory-based assessment of phosphorus and nitrogen loading from currently available commercial carp feeds

ABSTRACT: To clarify the total phosphorus (P) and nitrogen (N) loading from carp culture, five commercial diets were selected from a major location in Japan, Lake Kasumigaura, for a 12-week feeding trial. These diets were prepared as per the 'Kasumigaura Feed Standard' (crude protein < 35% and digestible energy > 3.5kcal/g), and total P ranged from 1.4% to 2.0%. However, for most of the diets the P available was lower than the requirement level. A control diet was formulated with 25% fishmeal to comply with that standard and contain adequate available P. Duplicate groups of juvenile carp were fed the aforementioned diets to satiation, three times a day, six days a week throughout the trial. Growth performance was significantly higher for the control group and values of P absorption (20.4–47.0%) and retention (14.0–36.3%) varied widely among the groups. Consequently, the total P loading (kg/t production) values based on retention fluctuated from 14.8 to 26.4 among the commercial diet groups compared with the low level of 8.5 for the control group. Similarly, the total N loading (kg/t production) values varied from 30.9 to 86.0 and was lowest for the control group. A higher whole body lipid and lower bone P and Ca confirmed the deficiency of the dietary available P in commercial diets. Better growth and comparatively less P and N loading rates were observed in the diet that had sufficient available P, not to mention that the control diet ranked best. It was concluded that an inadequacy of available P among the commercial diets affects the growth of carp and produces high P and N loading into the water. Therefore, if the commercial diets do not supply adequate levels of available P to carp, growth is negatively affected and may result in greater waste loading.     

<Emphasis Type="Italic">Cinderella</Emphasis> agroforestry systems

The underlying principles and traditional knowledge concerning indigenous agroforestry systems (AFS) around the world have been used successfully in the design of improved systems; but lately there has been a slow-down in that effort. Recognizing that Cinderella, by analogy, refers to an individual whose attributes were unrecognized or one who unexpectedly achieved success after a period of obscurity and neglect, we propose Cinderella agroforestry systems for such underexploited and “forgotten” AFS. In order to highlight their unrecognized potentials, a few such systems that are location-specific and little-known outside their areas of existence were selected from around the world. Each selected system is unique in terms of its production, environmental, and sociocultural attributes; but none had been described in quantitative terms of ecology and production. A qualitative SWOT (strengths–weaknesses–opportunities–threats) analysis of the systems showed several commonalities among them. While sustainability, multi-functionality, and high sociocultural values are common strengths, low levels of production and lack of research- and technology inputs to improve the system are the major weaknesses. The opportunities emanating from strengths and weaknesses are also common to all the systems; and “threats” to these systems arise mostly from ramifications of government policies. Given the acknowledged role of AFS in climate-smart agriculture and the current global momentum for working with communities to better manage landscapes, learning from success stories of the Cinderella AFS could suggest models for sustainable development, and efforts in that direction should also be incorporated into global agroforestry research agendas.     

Root activity of young <Emphasis Type="Italic">Acacia mangium</Emphasis> Willd trees: influence of stand density and pruning as studied by <Superscript>32</Superscript>P soil injection technique

Initial spacing and pruning are silvicultural strategies that influence the resource acquisition capabilities of trees. A field study was conducted in the humid tropics of peninsular India to test the assumptions that: (1) high stand density of Acacia mangium induces greater root uptake capacity close to the stem and from the subsoil; and (2) crown pruning stimulates greater root uptake capacity at proximal points. Root activity pattern of two-year-old A. mangium was evaluated as a function of three population densities (1,250, 2,500 and 5,000 stems ha⁻¹), with, and without 50% crown pruning, using ³²P soil injection. The label was placed at 25, 50 and 75 cm lateral distances and at 30 and 60 cm depth. Low density stands (1,250 stems ha⁻¹) generally showed higher ³²P recovery (P < 0.01), which was exaggerated by pruning. Pruned low density stands had 34% root activity at 25 cm, as against 23% for unpruned. The low density stands also showed higher root activity at 75 cm, signifying greater root spread. We suggest that high stem densities favour restricted spread of absorbing roots and may facilitate competitive downward displacement of roots. Pruning the lateral shoots at low stem densities may simulate this to some extent. The net outcome of interactions, however, will depend on trade-offs between stem density and tree management over time.