Protein quality of weaning foods based on locally available cereal and pulse combination

Locally available cereals and pulses such as rice (Oryza sativa), kangini (Setaria italica), sanwak (Echinochloa frumentacea), green gram (Vigna radiata) and jaggery were used to formulate three weaning foods. Cereal, pulse and jaggery were mixed in the ratio of 70:30:25. Roasting was the main processing technique used in the formulation of these weaning foods. The developed weaning foods had 5.06 to 5.68 g moisture, 10.28 to 13.71 g protein, 2.91 to 3.77 g ash, 1.08 to 1.87 g fat, 14.42 to 14.98 mg iron, 1.03 to 1.27 g crude fibre, and 357 to 374 Kcal. The weaning foods had a nutrient composition within the range prescribed by the Indian Standard Institute for processed weaning foods. The study indicated that the weaning foods obtained from locally available food stuffs have the potential of being produced locally, adaptable for household consumption and can be good substitute for commercial formulae.     

Improvement in HCl-extractability of minerals in home made weaning foods

Three weaning foods were formulated from locally available cereals and pulses such as rice (Oryza sativa), kangini (Setaria italica), sanwak (Echinochloa frumentacea), green gram (Vigna radiata) and jaggery. Cereals and pulses were mixed in the proportion of 7:3. Nutrient composition of developed weaning foods was within range prescribed by Indian Standard Institute and was found to be acceptable. Roasting was the processing technique employed in developing weaning foods which resulted in significant increase in HC1-extractable minerals, an index of their bioavailability to humans. The higher HC1-extractability of the minerals may be ascribed to the decreased phytic acid in the processed home made weaning foods.     

In infectious pancreatic necrosis virus carrier Atlantic salmon, Salmo salar L., post-smolts, almost all kidney macrophages ex vivo contain a low level of non-replicating virus

The level of infection by infectious pancreatic necrosis virus (IPNV) of kidney macrophages from 12 asymptomatic carrier Atlantic salmon post-smolts was studied. Kidney leucocytes were fractionated on 34/51% Percoll gradients, allowed to adhere to plastic wells overnight, washed to remove non-adherent cells and cultured for up to 7 days with or without renewal of medium on day 3. On day 1, supernatants were harvested, macrophages were counted, lysed and IPNV in the supernatants and lysates was titred in chinook salmon embryo (CHSE-214) cells. The multiplicity of infection ranged between 1:2.2 and 1:7.4 (virus:macrophages). On day 3, the titres of IPNV in macrophage lysates decreased and in wells where the medium was renewed on day 3, IPNV was no longer detectable on day 7. In the supernatants, one fish was positive for IPNV on day 1, four fish on day 3 but none were detectably positive on day 7. In parallel wells in which the medium was not renewed, on day 7 IPNV was detected in macrophage lysates of three fish and the supernatants were also IPNV positive in two of these fish. This suggests that virus might be shed from infected macrophages and then reinfect other macrophages. When macrophages were serially diluted in wells and cultured for 24 h, IPNV could be cultured from macrophage lysates of wells containing between two and 70 macrophages. These results indicate that a very high proportion of the adherent kidney macrophages must be infected with very few non-replicating virions.     

Protein and starch digestibilities and mineral availability of products developed from potato,soy and corn flour

A technique for development of potato flour was standardized. Five products viz. cake, biscuit, weaning food, panjiri and ladoo were prepared incorporating potato flour, defatted soy flour and corn flour. Baking and roasting were the major processing techniques employed for the development of these products. Protein, ash and fat contents of potato flour were almost similar to those of raw potatoes. Significant differences in protein, ash and fat contents of all the products were observed. Protein and starch digestibility of potato flour was significantly higher than that of raw potatoes. Protein digestibility increased by 12 to 17 percent on baking or roasting of products. Processed products had significantly higher starch digestibility and mineral availability compared to raw products. Thus, it can be concluded that roasting and baking are effective means of improving starch and protein digestibility and mineral availability of products.     

A sensitive loop-mediated isothermal amplification (LAMP) method for detection of Renibacterium salmoninarum, causative agent of bacterial kidney disease in salmonids

Loop-mediated isothermal amplification (LAMP) is a novel technique for nucleic acid amplification with high specificity, sensitivity and rapidity and does not require expensive equipment or reagents. In the present study, we developed and evaluated a LAMP method for the rapid detection of Renibacterium salmoninarum causing the bacterial kidney disease in salmonids. This method was more sensitive than quantitative real-time polymerase chain reaction (qPCR). Using DNA template extracted from cultured R. salmoninarum , the LAMP method gave an amplification signal from template diluted to 10⁻⁸ while the limit of detection of qPCR was10⁻⁷. The LAMP method was also highly specific and did not amplify DNA purified from five other Gram-positive and -negative bacterial fish pathogens. The method also worked well using extracts of macrophages infected with R. salmoninarum and kidney material from rainbow trout, which were positive for R. salmoninarum by qPCR and crude R. salmoninarum culture. There was some evidence for inhibitors of the LAMP reaction in the kidney samples, which was overcome by diluting the sample.     

A sensitive non-destructive method for detecting IPNV carrier Atlantic salmon, Salmo salar L., by culture of virus from plastic adherent blood leucocytes

In populations of Atlantic salmon in sea water, infectious pancreatic necrosis virus (IPNV) could be detected by standard virological culture methods in sonicated kidney homogenates and in mucus samples (gill, skin and rectum) from 14 and nine of 25 fish, respectively, but all fish were positive by virus culture from lysates of kidney macrophages and adherent blood leucocytes. In fish which tested negative for IPNV by the standard method of detection, the virus could be detected using adherent blood leucocytes isolated on a Percoll gradient from as little as 10 microL of blood. The blood sample could be stored for at least 3 days in a heparinized tube on ice before preparing the plastic adherent leucocytes. Furthermore, the latter could be prepared without prior fractionation on Percoll simply by incubating whole blood (33 microL) in cell culture medium (66 microL) in 96-well plates overnight and washing away the non-adherent cells before lysing the adherent cells and inoculation of the lysate onto CHSE-214 cells. This highly sensitive method for detecting IPNV-carriers is therefore very suitable for non-destructive sampling of fish in the field.     

Shelf life of weaning foods developed from locally available food stuffs

Four weaning foods were formulated using locally available cereals and pulses such as wheat (Triticum aestivum), barley (Hordeum vulgare), green gram (Vigna radiata) and jaggery. Cereal, pulse and jaggery were used in the proportion of 70:30:25. Roasting and malting were two processing techniques used. The developed weaning foods were evaluated for their nutritional characteristics and shelf life. All the formulations had a nutrient composition within the range prescribed by the Indian Standard Institute (ISI) for processed weaning foods. Peroxide value and fat acidity of weaning foods increased with increase in storage period. Malting of weaning foods resulted in higher increase of peroxide value and fat acidity as compared to roasted ones during the period of storage. All the blends were found to be acceptable up to 60 days of storage. The results, indicated that weaning foods developed from locally available less inexpensive foods may be used as good supplements for infants.     

Sequence and topological characterization of Toll-like receptor 8 gene of Indian riverine buffalo (Bubalus bubalis)

In this study, buffalo (Bubalus bubalis) Toll-like receptor 8 (TLR8) gene has been characterized by sequence analysis and detecting polymorphism. Complete ORF of buffalo TLR8 gene was amplified using the RNA isolated from spleen tissue, which was found to be 3,102 nucleotides long encoding a 1,033 amino acid protein. Buffalo TLR8 had 10 nucleotide changes as compared to other livestock species resulting in six unique amino acid changes, four of them lying within leucine-rich repeat (LRR) domains. As compared to cattle (Bos indicus and Bos taurus), out of fifteen cysteine residues, fourteen were conserved and Cys at position 521 was replaced by Arg. Nine of the LRR domains had no amino acid change as compared to cattle, whereas LRR-C-terminus had maximum, five amino acid changes. Sequence characterization of 12 riverine and swamp buffaloes revealed presence of four polymorphic nucleotides, two of them were non-synonymous, one synonymous and one site in 3′UTR. PCR-RFLP genotyping of non-synonymous SNP 2758A>G (ILeu920Val) in Toll–interleukin-1 receptor domain of 463 swamp and riverine buffaloes showed a higher frequency of allele A in swamp (95 %) as compared to riverine (9.84 %) buffaloes.