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.     

CO2 stunning of broilers and turkey hens

Stunning of poultry is still not solved satisfactorily. This concerns the requirements of animal welfare, meat quality and working conditions in the lairage, stunning and debleeding area. In an investigation of combined CO2-/O2-stunning in a new gas stunning system stress reactions of the animals during the induction phase and stunning effectivity were recorded in 7,000 chicken and 3.825 turkeys. During the induction phase (here: chicken 41 sek./turkeys 25-65 sec.) the animals first staid calm and then showed beak-opening as a consequence of the breathing stimulating effect of CO2. As a further sign of the aversiveness against CO2 the animals showed head shaking and wing flapping. The used settings of gas concentrations and stunning time in the system investigated lead to a very deep stunning resp. Killing of the animals. Therefore the debleeding cut could be performed late (chicken 44-55 sec., turkeys 54-90 sec.) after leaving the system without animals regaining consciousness. The stunning of turkeys with a mixture of CO2 and oxygen is an improvement according to animal welfare requirements because unnecessary pain and suffering, happening very often with electrical stunning, can be avoided. As far as animal welfare in chicken stunning is concerned it must be evaluated if gas stunning means an improvement because stress during the till now relatively long induction phase must be put into relation with comparably lower stress caused by hanging upside down and a fast and safe working electrical stunning unit.     

Erste Ergebnisse der Untersuchungen mit URUS-Mobilseilkran im Schwachholz

Ohne Zusammenfassung Diese Untersuchungen wurden von der Arbeitsgemeinschaft Deutscher Waldbesitzerverb?nde in dankenswerter Weise finanziell gef?rdert.     

Humane killing of animals in agricultural management]

The special problem of handling weak, chronically ill, injured farm animals or those not sellable for slaughter is described. Farmers generally do not have sufficient knowledge in killing animals. They need external help or they must take specific training courses to get the necessary knowledge. Solutions how to improve animal welfare in this field are given.     

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.     

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.     

Recommendations for future development in cattle transport in Europe

Every year millions of calves and cattle are transported across, from and to Europe. Most of these animals are going to slaughter houses in the respective countries or in another community state or coming or going abroad (extra-EU). These transports give cause for concern for at least three reasons: First, it can cause severe stress in animals entailing poor welfare. Second, stressful transports may have a negative effect on meat quality. Third, there is the risk of spread of infectious diseases over large distances. Existing legislation does not provide enough protection to transported animals especially over long distances largely because considerable parts of the regulations are not sufficiently based on scientific evidence. In recent years some research is carried out including the EU financed CATRA research project (contract QLK5-CT 1999-0157) concentrating on the welfare and meat quality aspects of cattle transport. This paper summarises important results of this recent research and gives some recommendations for future legislation. The welfare of the animals is limited by their needs not by a fixed maximum transport time, if vehicle and transport conditions are appropriate. Bulls, steers and heifers are reacting differently on transport. Adapt transport schemes to the needs of the animals. Meat quality is only effected in extreme situations. Some animals develop an energy deficit after 6 h of transport. Develop appropriate feeding regimes for long transport. Abolish stressful loading and unloading in staging posts (injuries, infectious diseases). Staging posts are particularly stressful for bulls. Educate handlers and drivers more intensively. Pay drivers inverse to losses. Develop monitor systems for long and short distance transport (e.g. records, GPS). Improve vehicle design (e.g. vibration).     

Detection of bicarbonate administration (milkshake) in Standardbred horses

SUMMARY Total plasma carbon dioxide (TCO₂) concentrations were measured in Standardbred horses to determine criteria to discriminate between normal horses and horses with excessive TCO₂ concentrations on raceday. TCO₂ concentrations from stabled horses were distributed normally with a mean of 30.2 mmol/L and a standard deviation of 1.2 (n = 192) while pre-race TCO₂ concentrations were not normally distributed. The results indicate that about 50 horses per million are likely to have TCO₂ concentrations greater than or equal to 35 mmol/L and that it is extremely unlikely that a normal horse would have a resting TCO₂ concentration above 36 mmol/L. These values were associated with sensitivities of 67% and 59%, respectively, and with a specificity of 100%. TCO₂ concentrations were relatively stable in blood samples stored at 4°C for 4 days, whereas the TCO₂ in specimens stored at room temperature (25°C) and at ambient temperature (16–28°C) declined progressively over 5 days. The accuracy and precision of the Beckman EL-ISE Auto Analyser were acceptable and within the manufacturers specified range. Paired specimens analysed using a Beckman EL-ISE Auto Analyser and a Kodak Dry Chemistry Analyser were not significantly different. However, the measurements made using the Kodak Dry Chemistry Analyser averaged 0.5 mmol/L higher than those analysed on the Beckman EL-ISE. The significance of these sources of variation in TCO₂ concentration in relation to the testing of horses for ‘milkshake’ administration are discussed.