Feline Ubiquitin Fusion Protein Genes

Using cDNA from a CRFK cell line as a template, PCR amplification was performed with the Ub1S and poly(dT) primers to isolate feline ubiquitin genes. Sequencing of the 495 bp PCR fragment revealed that the putative amino acids induced by this fragment gave a fusion protein consisting of a ubiquitin polypeptide (76 amino acids) and an extension protein of ribosomal proteins L40 (52 amino acids). The putative amino acid sequence of ubiquitin was identical to those of humans, rats and pigs.The recombinant glutathione S-transferase (GST)–feline ubiquitin fusion proteins were produced in Escherichia coli and purified. The fusion proteins had a molecular weight of about 42 kDa and were detected by immunoblot assay with rabbit anti-ubiquitin antiserum.The mRNAs from heat-shocked and non-heat-shocked cells were subjected to RT-PCR (Ub1S and poly(dT) primers) analysis. The molecular weights of the ubiquitinated proteins in heat-shocked CFRK cells were between 18 kDa and 24 kDa by immunoblot assay.These results suggested that there were more ubiquinated proteins in the heat-shocked CRFK cells than in the pre-heat-shocked cells.     

Characterisation of cross-reactivity of virus neutralising antibodies induced by feline panleukopenia virus and canine parvoviruses

It was recently reported that canine parvoviruses (CPV) had entered cat populations and induced disease in infected cats, while they had affected only dogs in the past. It is important to determine whether conventional feline panleukopenia virus (FPLV) vaccines protect against recent CPV infections. In this study, the cross-reactivity of virus-neutralising (VN) and haemagglutinin-inhibition (HI) antibodies in cats induced by FPLV and CPV s were examined. Lower cross-reactivities of VN and HI antibodies against each CPV strain were observed in cats experimentally inoculated with FPLV or vaccinated with an inactivated FPLV vaccine. In addition, we revealed the existence of a novel type of FPLV, which reacted weakly with antibodies induced by the conventional FPLV vaccine.     

Milk fever and subclinical hypocalcaemia--an evaluation of parameters on incidence risk, diagnosis, risk factors and biological effects as input for a decision support system for disease control.

The present review analyses the documentation on incidence, diagnosis, risk factors and effects of milk fever and subclinical hypocalcaemia. It is hereby evaluated whether the existing documentation seems sufficient for further modelling in a decision support system for selection of a control strategy. Several studies have been carried out revealing an incidence of milk fever most often in the level of 5-10%. Few studies indicate that the incidence of subclinical hypocalcaemia is several times higher than milk fever. The diagnosis based on clinical or laboratory methods or based on presence of risk factors is outlined. The clinical symptoms of milk fever are highly specific and the disease level may thus be determined from recording of treatments. Diagnosis of subclinical hypocalcaemia needs to include laboratory examinations or it may be determined by multiplying the incidence of milk fever by a certain factor. From the documentation on risk factors, it is very complex to predict the incidence from the exposure level of the risk factors. Due to uncertainty, sensitivity analyses over a wide range of values for each parameter are needed. The documentation of cow characteristics, nutrition, environment and management as risk factors are described. Among cow characteristics, parity or age, body condition and production level were found to be important. Risk factors associated with nutrition included most importantly dietary cation-anion difference and calcium level whereas the importance of general feeding related factors like type of feed stuff and feeding level were less clear. Environment and management included season, climate, housing, pasturing, exercise, length of dry period and prepartum milking. Several of the parameters on environment and management were confounded among each other and therefore firm conclusions on the importance were difficult. The documentation of the effect of milk fever includes the downer cows, reproductive disorders, occurrence of other diseases and the effect on milk production, body weight and culling. The reproductive disorders included most importantly dystocia, uterine prolapse, retained placenta, metritis and repeat breeding, and occurrence of other diseases included ketosis, displaced abomasum and mastitis. The documentation was substantial and often quantifiable within certain limits. Overall it is concluded that the present documentation on milk fever concerning incidence, diagnosis, risk factors and effects seems sufficient for a systematic inclusion in a decision support system. A model on milk fever should take into consideration the variation in biological data and individual herd characteristics. The inclusion of subclinical hypocalcaemia would be more uncertain and probably should await further documentation on possibilities of determining the herd level incidence and also the effect of this condition on production.     

A review of diet formulation strategies and feeding systems to reduce excretory and feed wastes in aquaculture

Significant efforts should be devoted to reducing waste outputs from aquaculture operations in order to lower the environmental impacts of aquaculture in many parts of the world. Since most aquaculture wastes are ultimately from dietary origin, reduction of waste outputs should first be through improvements of diet formulation and feeding strategies. The first step in the production of feeds producing less solid waste is to eliminate poorly digestible ingredients (such as whole grain or grain by-products used as binders and fillers in the feed formulae) and to use highly digestible ingredients with good binding properties. Further reduction of solid waste can then be achieved through careful selection of the ingredients to improve apparent digestibility and the nutrient balance of the feed. Nitrogen waste outputs can be reduced through the reduction of the digestible protein to digestible energy (DP/DE) ratio of the diet. Phosphorus waste outputs can be reduced through careful selection of the ingredients and optimization of the digestible phosphorus content of the diet to meet the requirement of the fish but avoid greatly exceeding this required level. Finally, feeding practices that minimize feed wastage should be adopted since feed wastage can have a very significant impact on waste outputs from fish culture operations.     

Localization of extracellular matrix receptors in ICGN mice, a strain of mice with hereditary nephrotic syndrome.

Fibrotic degeneration was examined in the kidneys of ICR-derived glomerulonephritis (ICGN) mice, a novel inbred mouse line with a hereditary nephrotic syndrome of unknown etiology considered to be a good model of human idiopathic nephrotic syndrome. In the present study, we histochemically revealed changes in accumulation of extracellular matrix (ECM) components and in localization of integrins, cellular receptors for ECM, in the kidneys of ICGN mice with the progression of renal failure. Excessive accumulation of basement membrane (laminin and collagen IV) and interstitial (type III collagen) ECM components were demonstrated in the glomeruli and tubulointerstitum of ICGN mice. Marked deposition of type I collagen and tenascin was seen only in the glomeruli of ICGN mice but not in those of ICR mice as normal controls. Increased expression of integrin alpha1-, alpha2-, alpha5- and beta1-subunits in glomeruli with fibrotic degeneration and abnormal distribution of alpha6-subunit were noted in the kidneys of ICGN mice. Excessive laminin, a ligand of alpha6beta1-integrin, was demonstrated on the tubular basement membrane, but alpha6-subunit diffusely disappeared on the basal side of the tubular epithelial cells. We presumed that abnormal integrin expression in renal tubules causes epithelial cell detachment, and consequently tubular nephropathy, and results in disorder of ECM metabolism causing excessive accumulation of ECM components in the kidneys of ICGN mice.     

Protection of turbot, Scophthalmus maximus (L.), and rainbow trout, Oncorhynchus mykiss (Richardson), against vibriosis using two different vaccines

Abstact. The potency of a whole-cell bacterin (WCB) and a toxoid enriched whole-cell vaccine (WCEB) administered intraperitoncally into rainbow trout, Oncorhynchus mykiss (Richardson), were compared. The most effective vaccine was further evaluated by bathing turbot, Scophthalmus maximus (L.). These vaccines were composed of three strains of V. anguillarun , of the serotypes 01 and 02. Both vaccines conferred the highest protection against strains of serotype 01 within 4 weeks. With the toxoid enriched vaccine giving the best results (77 RPS). When trout were revaccinated after 7 weeks with this vaccine, good protection was achieved against strains of serotypes 01 and 02. Interestingly, when the WCEB was administered by bath to turbot, acceptable levels of protection against strains of both serotypes were obtained after 4 weeks of immunization.     

Use of androgenesis in intersubspecific hybrid rice breeding

Significant differences of callus induction, green plant regeneration and culture efficiency were observed among restorer lines and their hybrids in rice. Average callus induction percentage of F₁, hybrids was about twice that of their parents. Twenty doubled haploid (DH) lines that showed normal fertility with both indica and japonica CMS lines, were selected as widely compatible restorers (WCRs). TG8 derived from CPSLO17/Minghui 63 not only showed normal fertility to both indica and japonica test varieties and CMS lines, but also exhibited superior agronomic traits, in particular short plant, desired plant type, high tillerability, large panicles, good grain quality and resistance to rice blast disease. Strong heterosis for yield was observed in crosses between indica or japonica CMS lines and TG8. TG8 possessed a dominant, widely compatible gene (WCG) that was inherited from the variety CPSLO17. Results confirmed that the anther culture technique is a quick and effective way of developing widely compatible restorers in rice and that it is applicable to the direct use of intersubspecific heterosis with the three-line method.