Proteomic profiling of plasma-derived small extracellular vesicles: a novel tool for understanding the systemic effects of tick burden in cattle

Cattle ticks pose a significant threat to the health and profitability of cattle herds globally. The investigation of factors leading to natural tick resistance in cattle is directed toward targeted breeding strategies that may combat cattle tick infestation on the genetic level. Exosomes (EXs), small extracellular vesicles (EVs) of 50 to 150 nm diameter, are released from all cell types into biofluids such as blood plasma and milk, have been successfully used in diagnostic and prognostic studies in humans, and can provide essential information regarding the overall health state of animals. Mass spectrometry (MS) is a highly sensitive proteomics application that can be used to identify proteins in a complex mixture and is particularly useful for biomarker development. In this proof of principle study, EXs were isolated from the blood plasma of cattle (Bos taurus) with high (HTR) and low tick resistance (LTR) (n = 3/group). Cattle were classified as HTR or LTR using a tick scoring system, and EXs isolated from the cattle blood plasma using an established protocol. EXs were subjected to MS analysis in data-dependent acquisition mode and protein search performed using Protein Pilot against the B. taurus proteome. A total of 490 unique proteins were identified across all samples. Of these, proteins present in all replicates from each group were selected for further analysis (HTR = 121; LTR = 130). Gene ontology analysis was performed using PANTHER GO online software tool. Proteins unique to HTR and LTR cattle were divided by protein class, of which 50% were associated with immunity/defense in the HTR group, whereas this protein class was not detected in EXs from LTR cattle. Similarly, unique proteins in HTR cattle were associated with B-cell activation, immunoglobins, immune response, and cellular iron ion homeostasis. In LTR cattle, unique exosomal proteins were associated with actin filament binding, purine nucleotide binding, plasma membrane protein complex, and carbohydrate derivative binding. This is the first study to demonstrate that MS analysis of EXs derived from the blood plasma of HTR and LTR cattle can be successfully applied to profile the systemic effects of tick burden.     

Immune response of chicks to oral vaccination against Newcastle disease and fowl pox

The humoral immune response and immunity conferred in chicks were compared following separate and combined oral vaccination with F strain of Newcastle disease virus (NDV) and HP1 strain of fowl pox virus. The haemagglutination inhibition (HI) antibody titre against NDV and passive haemagglutination (PHA) antibody titre against fowl pox virus were comparable in two respective groups. The serum IgG concentration increased significantly after the second vaccination in all the groups. The NDV vaccine induced significantly higher IgG production as compared to fowl pox virus vaccine. There was no significant difference in serum IgG concentration produced by combined vaccine and separate F strain vaccine. The protection afforded by combined and separate vaccinations did not vary significantly against challenge with virulent strains of NDV and fowl pox virus at different stages.     

Romifidine-ketamine anaesthesia in atropine and triflupromazine pre-medicated buffalo calves

The study was conducted on 10 buffalo calves with a weight of 98.5 +/- 3.9 kg and age 9.7 +/- 1.3 months. Ten trials of two treatments were carried out using a randomized block design. Atropine at the dose of 0.02 mg/kg bodyweight was administered in both the groups. The animals of group I received romifidine at the dose of 10 microg/kg i.v., 10 min after atropine administration, whereas, animals of group II received triflupromazine at the dose of 0.3 mg/kg i.m. and 10 min later romifidine at the dose of 10 microg/kg i.v. immediately followed by ketamine at the dose of 5 mg/kg i.v. The onset of action of romifidine in group I occurred within 2 min and the animals remained under mild sedation for 31 +/- 4.8 min. In group II, the triflupromazine-romifidine-ketamine combination induced anaesthesia for 14 +/- 2.3 min. Hypothermia, significant bradycardia and respiratory depression was noticed in both groups at different time intervals.     

Xylazine, diazepam and midazolam premedicated ketamine anaesthesia in white Leghorn cockerels for typhlectomy

Thee different combinations of ketamine hydrochloride were used to induce general anaesthesia for surgical operations (typhlectomy) in 30 adult, single-comb White Leghorn cockerels. They were randomly divided into three groups, each comprising 10 birds. Birds in Group I received xylazine-ketamine combinations at the dose rate of 2 mg xylazine and 10 mg ketamine per kg i.v., whereas birds of Group II received diazepam (2.5 mg/kg i.v.) and 5 min later ketamine (75 mg/kg i.m.). In the Group III, midazolam (2 mg/kg i.m.) and 5 min later ketamine (50 mg/kg i.v.) was administered. The onset of sedation/anaesthesia was shortest (1.60 +/- 0.27 min) in Group I, followed by Group II (8.40 +/- 0.83 min) and Group III (17.10 +/- 1.71 min). Recovery period was shortest in the Group I (65-75 min) followed by Group II (80-85 min) and Group III (92-105 min). Sedation, muscle relaxation and surgical anaesthesia was optimal and excellent in Group I compared with the other two groups. Torticollis, salivation and dyspnoea were observed in Group III. Short-term limb contractions were present in all birds in Groups II and III, up to 20 min of observation. Recovery from anaesthesia was smooth in all three groups. A Surgical procedure (typhlectomy) was performed on all birds. Hypothermia was observed in Group II, whereas heart and respiratory depression was recorded in Group I. Blood sugar level did not vary significantly in any anaesthetic regime. The reduction of haemoglobin was maximum in Group II compared with Groups I and III. Hypoxaemia and hypercapnaea were elevated in all birds in Groups II and III. Blood electrolytes did not vary significantly from the baseline values among the three groups of birds during the period of observation (120 min). The xylazineketamine combination was found to be the best anaesthesia for surgical intervention in chickens.     

新疆药桑组培育苗的方法与培养基筛选试验研究

新疆药桑的组培育苗需要经过初代培养、继代培养、生根培养、驯化移栽几个阶段。经试验比较筛选出了适合不同阶段生根发苗的培养基配方,并阐述了新疆药桑组培快繁的方法和步骤。     

Genotyping of Edwardsiella tarda isolated from freshwater fish culture system

The applicability of PCR-RFLP of 16S rDNA and conventional phenotypic methods for differentiation of Edwardsiella tarda associated in freshwater fish culture system was studied. In this study, by conventional biochemical tests and antibiotic resistant patterns 2 and 14 groups were obtained. But these methods failed to discriminate the isolates habitat wise. However, PCR-RFLP of 16S rDNA was found to be specific to detect habitat-specific isolates. All the fish isolates belonging to particular genotypes were found only in fish, not in water or sediment. Some of the genotypes were exclusively present in water and sediment. This study indicates the prevalence of site-specific genotypes in freshwater ecosystems. Molecular method is found to be superior to discriminate the E. tarda habitat wise to conventional typing methods.     

Karanj protein isolate prepared from karanj seed cake: Effect on growth,body composition and physiometabolic responses in Labeo rohita fingerlings

A protein isolate was prepared from karanj seed (KPI) with 921.2 g protein/kg seed, which contained a negligible amount of anti‐nutritional factors and a balanced amino acid composition, especially rich in methionine. For 60‐day feeding trial, five isonitrogenous (300 g/kg CP) and isocaloric (15 MJ DE/kg) diets were formulated by replacing soybean protein isolate (SPI) on protein equivalent basis, KPI‐0 (control, 0 g/kg KPI); KPI‐25 (replacing 250 g/kg SPI protein with KPI); KPI‐50 (replacing 500 g/kg SPI protein with KPI); KPI‐75 (replacing 750 g/kg SPI protein with KPI) and KPI‐100 (replacing 1,000 g/kg SPI protein with KPI) for the feeding of L. rohita. The weight gain percentage, specific growth rate, feed conversion ratio and protein efficiency ratio were not significantly (p > .05) varied among the KPI fed and control groups. A significantly higher hepatosomatic index was recorded in the control and KPI‐25 groups compared with other groups. The whole‐body compositions, except ether extract, did not differ significantly (p > .05) among the groups. Digestive (amylase, protease, lipase and alkaline phosphatase) and metabolic enzyme activities (hexokinase, transaminases and lactate dehydrogenase) and glycogen stores were not significantly affected, whereas intestinal alkaline phosphatase differed significantly (p < .05). The RNA–DNA ratio was significantly (p < .05) higher in the KPI‐75 group. Thus, the study revealed that KPI can completely replace SPI protein at 191 g/kg inclusion level in the diets of L. rohita fingerlings.