Identification and validation of cold responsive microRNAs of <Emphasis Type="Italic">Hevea brasiliensis</Emphasis> using high throughput sequencing

Cold stress is one of the major abiotic factors that influence the productivity and geographical distribution of many agriculturally important crops like Hevea brasiliensis. Cultivation of H. brasiliensis in India is being extended to northeastern regions, where low temperature during winter adversely affects its survival, growth, and productivity. Developing cold-tolerant genotypes is a primary requisite to maximize the productivity under such challenging environmental conditions. However, lack of methods for early evaluation of cold tolerance in the newly developed clones and the extensive time required for assessing their tolerance in the field are major constraints for clonal selection. The present study was initiated with an objective to identify and characterize cold stress responsive miRNAs from H. brasiliensis that show stronger association with cold tolerance. Next generation sequencing using Illumina HiSeq method revealed the expression of 21 and 29 conserved miRNA (from clone RRIM 600) families in cold-stressed and control samples, respectively. Forty-two novel miRNAs were identified from this study. Upon differential expression analysis, eight conserved miRNAs were found commonly expressed in both the samples. When expression analyses were performed subsequently with six selected miRNAs in two Hevea clones (viz. RRII 105 and RRIM 600), miR169 showed a strong association with cold tolerance. miRNAs such as miR482 and miR159 also exhibited association with cold tolerance. This study suggests the possibility of employing these miRNAs as markers for cold tolerance after validation in more number of genotypes with varying levels of cold tolerance.     

Problems and Programs in the Application of the Sterility Principle for the Control of Ceratitis capitata Wied.

In spite of the good results obtained in the control of Ceratitis under field conditions on the Italian islands of Capri and Procida, in Nicaragua and in the Spanish Murcia area, several problems connected with the sterile male technique still have to be solved. Mass rearing of the insects must be improved. Perhaps, total sterility of the males is not required and the necessary dose of irradiation must be studied in relation to its effect on their competitiveness in the field. In checking the efficacy of field operations the recapture of marked individuals will have to be supplemented or gradually replaced by the time-consuming assessment of egg-hatching and fruit-damage. Studies of the influence of environmental factors on population densities must be intensified.     

Formation of Biogenic Amines and Associated Biochemical and Microbial Attributes of Whole Sutchi Catfish (Pangasianodon hypophthalmus) during Iced Storage

Sutchi catfish (Pangasianodon hypophthalmus) is a highly preferred farmed species that is produced in huge quantities. Meat from ice-stored (4 ± 2°C) whole Sutchi catfish was evaluated for formation of biogenic amines, such as putrescine, cadaverine, histamine (HIM), agmatine, tyramine, spermine, and spermidine, and compared with biochemical, microbial, and sensory attributes during 22 days. Analysis of content of biogenic amines in the meat was carried out by liquid chromatography with tandem mass spectrometry using a without derivatization method. Three amines, namely, tyramine, spermidine, and spermine, were only present on 0^th day of storage. Presence of cadaverine was noticed from 9^th day onwards, where the aerobic plate count (APC) reached 4.85 log CFU/g. Putrescine was detected on the 22^nd day of storage, where the APC crossed 7 log CFU/g. HIM was detected in lower quantities from 3^rd day onwards. A shelf life of 15 days was determined based on sensory and microbiological evaluation. Although the samples were biochemically acceptable throughout the storage period, APC exceeded the limit on day 19, and the gradual increase of H₂S-producing bacteria, Brochothrix thermosphacta, Aeromonas, and Enterobacteriaceae, was observed during storage.     

Effect of repeated exposure to AQUI‐S® on the viability and growth of Neoparamoeba perurans

There have been recent efforts amongst immunologists to develop approaches for following individual fish during challenges with viral and bacterial pathogens. This study contributes to assessing the feasibility of using such approaches to study amoebic gill disease (AGD). Neoparamoeba perurans, agent of AGD, has been responsible for widespread economic and fish loss in salmonid aquaculture. With the emergence of AGD in Europe, research into infection dynamics and host response has increased. This study investigated the effect of repeat exposure to anaesthesia, a necessary requirement when following disease progression in individual fish, on N. perurans. In vitro cultures of N. perurans were exposed every 4 days over a 28‐day period to AQUI‐S^® (isoeugenol), a popular anaesthetic choice for AGD challenges, at a concentration and duration required to sedate post‐smolt salmonids. Population growth was measured by sequential counts of amoeba over the period, while viability of non‐attached amoeba in the culture was assessed with a vital stain. AQUI‐S^® was found to be a suitable choice for in vivo ectoparasitic challenges with N. perurans during which repetitive anaesthesia is required for analysis of disease progression.     

Expression of kit ligand and insulin‐like growth factor binding protein 3 during in vivo or in vitro development of ovarian follicles in sheep

Expression of Kit ligand (KL) and insulin‐like growth factor binding protein (IGFBP3) genes was studied at different in vivo and corresponding in vitro stages of development of the ovarian follicles in sheep. The expression of both KL and IGFBP3 was significantly higher in the primordial follicles relative to any other stage of development. Compared to the other stages, the KL expression in the cumulus cells from in vivo grown large antral follicles and that of IGFBP3 in COCs’ isolated from large antral follicles matured in vitro for 24 hr were significantly higher. In the oocytes from in vivo grown ovarian follicles, the expression of KL was the same at all the stages of development. Insulin‐like growth factor binding protein 3 expression was also the same in the oocytes at all the stages of the development except for a significantly lower expression in those from antral follicles. The expression of KL in the cumulus cells decreased significantly in the in vitro grown early antral follicles but did not change further as the development progressed. The expression of IGFBP3 in the cumulus cells from in vitro grown ovarian follicles appeared to increase as the development progressed although the increase was not significant between any two consecutive stages of development. In the oocytes in in vitro grown ovarian follicles, the expression levels of KL and IGFBP3 genes did not change with development. It is concluded that (i) KL and IGFBP3 genes follow specific patterns of expression during ovarian folliculogenesis and (ii) in vitro culture of preantral follicles compromises the development potential through alterations in the stage‐specific patterns of expression of these and other developmentally important genes.