A heterologous expression system for bovine lens transmembrane main intrinsic protein (MIP) in Nicotiana tabacum plants

We have developed a heterologous expression system for transmembrane lens main intrinsic protein (MIP) in Nicotiana tabacum plant tissue. A native bovine MIP26 amplicon was subcloned into an expression cassette under the control of a constitutive Cauliflower Mosaic Virus promoter, also containing a neomycin phosphotransferase operon. This cassette was transformed into Agrobacterium tumefaciens by triparental mating and used to infect plant tissue grown in culture. Recombinant plants were selected by their ability to grow and root on kanamycin-containing media. The presence of MIP in the plant tissues was confirmed by PCR, RT-PCR and immunohistochemistry. A number of benefits of this system for the study of MIP will be discussed, and also its application as a tool for the study of heterologously expressed proteins in general.

Farm costs associated with premovement testing for bovine tuberculosis

Sixty cattle farmers in England were questioned about the costs associated with premovement testing for bovine tuberculosis (TB). On average, the farmers had premovement tested 2-45 times in the previous 12 months, but the majority had tested only once. An average of 28.6 animals were tested on each occasion, but there were wide variations. The average farm labour costs were (sic)4.00 per animal tested, veterinary costs were (sic)4.33 and other costs were (sic)0.51, giving a total cost of (sic)8.84, but there were wide variations between farms, and many incurred costs of more than (sic)20 per animal. A majority of the farmers also cited disruption to the farm business or missed market opportunities as costs, but few could estimate their financial cost. Most of the farmers thought that premovement testing was a cost burden on their business, and over half thought It was not an effective policy to control bovine TB.

Economic assessment of bovine tuberculosis and alternative policies

Break-even analyses of the costs and benefits of six alternative bovine tuberculosis (bTB) control strategies were undertaken. The results show that some strategies, such as zoning, would require relatively small reductions in bTB incidence as a result to be cost effective, whilst for others, such as proactive badger removal, the costs would require a substantial and relatively rapid reduction in bTB incidence to be worthwhile.

Public values for badgers, bovine TB reduction and management strategies

This paper applies an attribute-based stated choice experiment approach to estimate the value that society places on changes to the size of the badger population in England and Wales. The study was undertaken in the context of a rising incidence of bovine tuberculosis (bTB) in cattle and the government's review of current bTB control policy. This review includes consideration of culling badgers to reduce bTB in cattle, since badgers are thought to be an important wildlife reservoir for the disease. The design of the CE involved four attributes (size of badger population, cattle slaughtered due to bTB, badger management strategy and household tax) at four levels with eight choice sets of two alternatives presented to respondents. Telephone interviews were undertaken with over 400 respondents, which elicited their attitudes and preferences concerning badgers, bTB in cattle and badger management strategies. The study estimated a willingness to pay of 0.10 pound per household per year per 100,000 badgers and 1.52 pound per household per year per 10,000 cattle slaughtered due to bTB which aggregated to 22 per badger and 3298 pound per bTB slaughtered animal for all households in England and Wales. Management strategy toward badgers had a very high valuation, highlighting the emotive issue of badger culling for respondents and the importance of government policy towards badgers.

Dietary supplements of whole linseed and vitamin E to increase levels of alpha-linolenic acid and vitamin E in bovine milk

The potential to increase the concentrations of n-3 polyunsaturated fatty acids (PUFAs) in milk fat was investigated by studying the effects of feeding a xylose-treated, whole cracked linseed supplement ( rich in alpha-linolenic acid) to dairy cows. Also the effect of increasing the dietary intake of vitamin E on the vitamin E status of milk was investigated. The effect of pasteurisation on milk fatty acid composition was also examined. Using a 3 x 2 factorial design, a total of 60 Holstein dairy cows were fed a total mixed ration based on grass silage supplemented with one of three levels of whole cracked linseed (78, 142 or 209 g . kg(-1) diet dry matter (DM); designated LL, ML or HL, respectively) in combination with one of two levels of additional dietary vitamin E intake ( 6 or 12 g vitamin E . animal(-1) . day(-1); designated LE or HE, respectively). Increasing lipid supplementation reduced (P < 0.01) diet DM intake and milk yield, and increased (P < 0.001) the overall content of oleic, vaccenic, alpha-linolenic and conjugated linoleic acids, and total PUFAs and monounsaturated fatty acids (MUFA). Myristic and palmitic acids in milk fat were reduced ( P < 0.001) through increased lipid supplementation. While α-linolenic acid concentrations were substantially increased this acid only accounted for 0.02 of total fatty acids in milk at the highest level of supplementation (630 g α-linolenic acid &BULL; animal(-1) &BULL; day(-1) for HL). Conjugated linoleic acid concentrations in milk fat were almost doubled by increasing the level of lipid supplementation (8.9, 10.4 and 16.1 g &BULL; kg(-1) fatty acids for LL, ML and HL, respectively). Although milk vitamin E contents were generally increased there was no benefit (P > 0.05) of increasing vitamin E intake from 6 to 12 g . animal(-1) . day(-1). The fatty acid composition of milk was generally not affected by pasteurisation.

Enhancement of oleic acid and vitamin E concentrations of bovine milk using dietary supplements of whole rapeseed and vitamin E

With the aim of reducing the degree of saturation and increasing the C18:1 cis fatty acid content of milk fat, the effects of feeding high levels of whole cracked rapeseed to dairy cows was investigated together with the effect of increasing dietary intake of vitamin E on the vitamin E content of milk. Using a 3 x 3 factorial design, 90 Holstein dairy cows were fed one of three levels of whole cracked rapeseed (0 (ZR), 134 (MR) and 270 g . kg(-1) diet dry matter (DM) (HR)) in combination with one of three intakes of supplementary vitamin E (0 (ZE), 2 (ME) and 4 g . cow(-1) . d(-1) (HE)). Supplementing with up to almost 2 kg . d(-1) of rapeseed oil (diet HR) significantly (P < 0.001) increased C18: 1cis in milk fat, from 181 (ZR) to over 400 g &BULL; kg(-1) (HR) of total milk fatty acids. Concentrations of C18: 0, C18: 2 and C18: 3 fatty acids were also increased ( P < 0.001) but by a much lesser degree, and the saturated fatty acids C4: 0 to C16: 0 decreased substantially. Vitamin E supplementation increased ( P < 0.01) milk vitamin E concentrations from 1.29 (ZE) to 1.68 mg &BULL; kg(-1) whole milk (HE). Thus substantial changes in milk fat composition with potentially beneficial effects on human health were achieved and without any adverse effects on milk taste. However, these improvements must be offset against the substantial reductions ( P < 0.001) observed in voluntary feed DM consumption (ZR, 20.6; HR, 15.2 kg DM . d(-1)), milk yield (ZR, 22.9; HR, 13.2 kg . d(-1)) and milk fat concentration (ZR, 42.1; HR, 33.4 g . kg(-1)) which would not be commercially sustainable unless a considerable premium was paid for this modified milk. It seems likely that the optimum dose of dietary rapeseed is lower than used in this study.

Enhancing the selenium content of bovine milk through alteration of the form and concentration of selenium in the diet of the dairy cow

Since estimated dietary selenium intake in the UK has declined steadily from around 60 mug day(-1) in 1975 to 34 mug day(-1) in 1997, there is a need to increase selenium intake from staple foods such as milk and milk products. An experiment was therefore done to investigate the relationship between dietary source and concentration of selenium and the selenium content of bovine milk. In a 3 x 3 factorial design, 90 mid-lactation Holstein dairy cows were supplemented over 8 weeks with either sodium selenite (S), a chelated selenium product (Selenium Metasolate(TM)) (C) or a selenium yeast (Sel-plex(TM)) (Y) at three different dietary inclusion levels of 0.38 (L), 0.76 (M) and 1.14 (H) mg kg(-1) dry matter (DM). Significant increases in milk selenium concentration were observed for all three sources with increasing inclusion level in the diet, but Y gave a much greater response (up to +65 mug l(-1)) than the other two sources of selenium (S and C up to +4 and +6 mug l(-1) respectively). The Y source also resulted in a substantially higher apparent efficiency of transfer of selenium from diet to milk than S or C. Feeding Y at the lowest dietary concentration, and thus within the maximum level permitted under EU regulations, resulted in milk with a selenium concentration of 28 mug l(-1). If the selenium concentration of milk in the UK was increased to this value, it would, at current consumption rates, provide an extra 8.7 mug selenium day(-1), or 11 and 14% of daily recommended national intake for men and women respectively. (C) 2004 Society of Chemical Industry.

Effect of replacing grass silage with maize silage in the diet on bovine milk fatty acid composition

Even though extensive research has examined the role of nutrition on milk fat composition, there is less information on the impact of forages on milk fatty acid (FA) composition. In the current study, the effect of replacing grass silage (GS) with maize silage (MS) as part of a total mixed ration on animal performance and milk FA composition was examined using eight multiparous mid-lactation cows in a replicated 4 X 4 Latin square with 28-day experimental periods. Four treatments comprised the stepwise replacement of GS with MS (0, 160, 334 and 500 g/kg dry matter (DM)) in diets containing a 54:46 forage: concentrate ratio on a DM basis. Replacing GS with MS increased (P < 0.001) the DM intake, milk yield and milk protein content. Incremental replacement of GS with MS in the diet enhanced linearly (P < 0.001) the proportions of 6:0-14:0, decreased (P < 0.01) the 16:0 concentrations, but had no effect on the total milk fat saturated fatty acid content. Inclusion of MS altered the distribution of trans-18:1 isomers and enhanced (P < 0.05) total trans monounsaturated fatty acid and total conjugated linoleic acid content. Milk total n-3 polyunsaturated fatty acid (PUFA) content decreased with higher amounts of MS in the diet and n-6 PUFA concentration increased, leading to an elevated n-6: n-3 PUFA ratio. Despite some beneficial changes associated with the replacement of GS with MS, the overall effects on milk FA composition would not be expected to substantially improve long-term human health. However the role of forages on milk fat composition must also be balanced against the increases in total milk and protein yield on diets containing higher proportions of MS.

Evaluation of PCR to detect Theileria parva in field-collected tick and bovine samples in Tanzania

The ability of PCR to detect infections of Theileria parva, the cause of East Coast Fever, in field-collected tick and bovine samples from Tanzania was evaluated. PCR-detected infection prevalence was high (15/20, 75%) in unfed adult Rhipicephalus appendiculatus ticks that fed as nymphs on an acutely-infected calf, but low (22/836, 2.6%) in unfed adult R. appendiculatus collected from field sites in Tanzania. Tick infection prevalence was comparable to that in previous studies that used salivary gland staining to detect T parva infection in field-collected host-seeking ticks. Of 282 naturally-exposed zebu calves, seven had PCR-positive buffy coat samples prior to detection of Theileria spp. parasites in stained huffy coat cells or lymph node biopsies. Evidence of Theileria spp. infections was detected in stained smears of lymph node biopsies from 109 calves (38.6%) and huffy coat samples from 81 (28.7%), while huffy coat samples from 66 (23.4%) were PCR-positive for T parva. Implications of these findings for the sensitivity and specificity of the PCR are discussed. (C) 2003 Elsevier Science B.V. All rights reserved.

A cost-benefit analysis of culling badgers to control bovine tuberculosis

Bovine tuberculosis (TB) is an important economic problem. The incidence of TB in cattle herds has steadily risen in the UK, and badgers are strongly implicated in spreading disease. Since the mid-1970s the UK government has adopted a number of badger culling strategies to attempt to reduce infection in cattle. In this report, an established model has been used to simulate TB in badgers, transmission to cattle, and control by badger culling. Costs were supplied by the UK Government's Department for Environment Food and Rural Affairs (Defra) for badger trapping and gassing. Regardless of culling intensity or area simulated, an overall reduction in the herd breakdown rate was seen. With a high culling efficacy and no social perturbation, the mean Net Present Value of a few simulated culling strategies in an "ideal world" was positive, meaning the economic benefits outweighed the costs. Further work is required before these results could be considered definitive, as it is necessary to evaluate uncertainties and simulate less than perfect conditions. (c) 2005 Elsevier Ltd. All rights reserved.

A breakthrough in lupin biotechnology: prolific protocolonisation in recalcitrant white lupin (Lupinus albus) triggered by bovine serum albumin

Six nutrient formulations were studied for their efficacy in inducing mitosis in white lupin seedling cotyledon protoplasts of which the formulations of Schafer-Menuhr & Sturmer (AS) and Kao (K8p) were found to be superior over the other four when supplemented with 6-benzylaminopurine and alpha-naphthaleneacetic acid (alpha-NAA). An unltrafiltration treatment of K8p increased mitotic frequency by 130% when compared with the untreated control. Medium enrichment with 0.2% bovine serum albumin (BSA) brought about a dramatic 1341% rise in protoplast division in comparison with BSA-free medium but only when the enrichment was carried out in Kao and Michayluk (KM8p) background containing 2, 4-dichlorophenoxyacetic acid, alpha-NAA and zeatin. A higher number of protocolonies (each proliferating from single protoplast following multiple divisions) were seen in 0.4% BSA. With this breakthrough in white lupin protoplast research, it is now possible to reproducibly obtain protocolonies that was hitherto not possible.

Viral nucleolar localisation signals determine dynamic trafficking within the nucleolus

Localisation of both viral and cellular proteins to the nucleolus is determined by a variety of factors including nucleolar localisation signals (NoLSs), but how these signals operate is not clearly understood. The nucleolar trafficking of wild type viral proteins and chimeric proteins, which contain altered NoLSs, were compared to investigate the role of NoLSs in dynamic nucleolar trafficking. Three viral proteins from diverse viruses were selected which localised to the nucleolus; the coronavirus infectious bronchitis virus nucleocapsid (N) protein, the herpesvirus saimiri ORF57 protein and the HIV-1 Rev protein. The chimeric proteins were N protein and ORF57 protein which had their own NoLS replaced with those from ORF57 and Rev proteins, respectively. By analysing the sub-cellular localisation and trafficking of these viral proteins and their chimeras within and between nucleoli using confocal microscopy and photo-bleaching we show that NoLSs are responsible for different nucleolar localisations and trafficking rates.

The postfusion structure of baculovirus gp64 supports a unified view of viral fusion machines

Viral fusion proteins mediate the merger of host and viral membranes during cell entry for all enveloped viruses. Baculovirus glycoprotein gp64 (gp64) is unusual in promoting entry into both insect and mammalian cells and is distinct from established class I and class II fusion proteins. We report the crystal structure of its postfusion form, which explains a number of gp64's biological properties including its cellular promiscuity, identifies the fusion peptides and shows it to be the third representative of a new class (III) of fusion proteins with unexpected structural homology with vesicular stomatitis virus G and herpes simplex virus type 1 gB proteins. We show that domains of class III proteins have counterparts in both class I and II proteins, suggesting that all these viral fusion machines are structurally more related than previously thought.