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The Pirbright Institute publication directory contains details of selected publications written by our researchers.

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Limon G, Lewis E G, Chang Y-M, Ruiz H, Balanza M E, Guitian J (2014)

Using mixed methods to investigate factors influencing reporting of livestock diseases: A case study among smallholders in Bolivia

Preventive Veterinary Medicine 113 (2), 185-196
Publisher’s version: https://doi.org/10.1016/j.prevetmed.2013.11.004

Abstract

Livestock disease surveillance is particularly challenging in resource-scarce settings, where disease events are often unreported. Surveillance performance is determined as much by the quantifiable biological attributes of the disease, as it is by motivations and barriers perceived by livestock keepers for disease reporting. Mixed methods designs, which integrate the collection, analysis and interpretation of qualitative and quantitative data in a single study, are increasingly used across different disciplines. These designs allow for a deeper exploration of the topic under investigation, than can be achieved by either approach alone. In this study a mixed methods design was used in order to gain a greater understanding of the factors that influence reporting of livestock diseases in Bolivia. There is a need to strengthen passive surveillance in this country, among other reasons as part of an eradication programme for Foot and Mouth Disease (FMD). Findings revealed livestock keepers in the study area were extremely unlikely to report the occurrence of livestock health events to the Official Veterinary Services (OVS). Communication outside the local community occurs more often through alternative routes and this is positively correlated with disease awareness. The main barriers to disease reporting identified were a lack of institutional credibility and the conflicting priorities of the OVS and livestock keepers. As for other animal and human diseases across the developing world, passive surveillance of livestock diseases in Bolivia should be enhanced; this is urgent in view of the current FMD eradication programme. Increasing timeliness and smallholders’ participation requires a detailed understanding of their likely actions and perceived barriers towards disease reporting. These insights are most likely to be developed through a holistic mixed methods approach of quantitative and qualitative analyses.

Rooney A L, Limon G, Vides H, Cortez A, Guitian J (2014)

Sarcocystis spp. in llamas (Lama glama) in Southern Bolivia: A cross sectional study of the prevalence, risk factors and loss in income caused by carcass downgrades

Preventive Veterinary Medicine 116 (3), 296-304
Publisher’s version: https://doi.org/10.1016/j.prevetmed.2013.11.014

Abstract

Llamas (Lama glama) are intermediate hosts of the protozoan parasite Sarcocystis spp. This parasite is described as causing economic losses in the production of llama meat in South America. The aim of this study was to estimate prevalence, identify risk factors and explore spatial patterns of Sarcocystis in llamas in an area of the Bolivian High Plateau including estimating financial losses due to carcass downgrades as a result of the presence of Sarcocystis cysts. Information was collected from a local abattoir between 2006 and 2011 on 1196 llamas. Sarcocystis status was determined at meat inspection where any carcasses with one or more visible cysts were deemed Sarcocystis positive. A high prevalence was found, estimated to vary between 23.4% (95% CI 16.6–30.1) in 2007 and 50.3% (95% CI 44.4–56.3) in 2011. Period prevalence between 2006 and 2011 was estimated at 34.1% (95% CI 31.4–36.8). Age, sex and type (analogous to breed) were identified as risk factors for Sarcocystis using a mixed-effects logistic regression model adjusting for clustering by community and owner. Llamas over 4.5 years of age had an increased odds of being Sarcocystis positive (OR 19.31, 95% CI 9.10–40.98) as well as females (OR 1.75, 95% CI 1.13–2.68) and long haired type llamas (OR 1.90, 95% CI 1.26–2.87). An interaction between age and sex was detected indicating that the increased odds of disease from the youngest age group to the 2.5–4.5 years group was much more pronounced in females than in males. Spatial patterns of Sarcocystis were explored at district level by means of Standardised Morbidity Ratios and some spatial heterogeneity was revealed. Estimates of financial loss due to the disease were calculated using the difference in price paid for Sarcocystis positive and negative meat. Loss due to Sarcocystis varied per year but could be up to 20% of the annual income generated through the abattoir by sale of meat. Overall this study shows a high prevalence of Sarcocystis in the study area with some heterogeneity between districts. It also identifies some previously unknown risk factors for Sarcocystis and gives financial estimates of the cost of the disease as a result of carcass downgrades. We hoped these findings will add to the understanding of Sarcocystis in llamas in Southern Bolivia and will be useful when considering if controls are necessary, worthwhile and practical.

Chung L, Bailey D, Leen E N, Emmott E P, Chaudhry Y, Roberts L O, Curry S, Locker N, Goodfellow I G (2014)

Norovirus translation requires an interaction between the C Terminus of the genome-linked viral protein VPg and eukaryotic translation initiation factor 4G

Journal of Biological Chemistry 289 (31), 21738-50
Publisher’s version: http://dx.doi.org/10.1074/jbc.M114.550657

Abstract

Viruses have evolved a variety of mechanisms to usurp the host cell translation machinery to enable translation of the viral genome in the presence of high levels of cellular mRNAs. Noroviruses, a major cause of gastroenteritis in man, have evolved a mechanism that relies on the interaction of translation initiation factors with the virus-encoded VPg protein covalently linked to the 5' end of the viral RNA. To further characterize this novel mechanism of translation initiation, we have used proteomics to identify the components of the norovirus translation initiation factor complex. This approach revealed that VPg binds directly to the eIF4F complex, with a high affinity interaction occurring between VPg and eIF4G. Mutational analyses indicated that the C-terminal region of VPg is important for the VPg-eIF4G interaction; viruses with mutations that alter or disrupt this interaction are debilitated or non-viable. Our results shed new light on the unusual mechanisms of protein-directed translation initiation.

Goodfellow I, Bailey D (2014)

Detection of protein-protein interactions using tandem affinity purification

Methods in Molecular Biology 1177, 121-33
Publisher’s version: http://dx.doi.org/10.1007/978-1-4939-1034-2_10

Abstract

Tandem affinity purification (TAP) is an invaluable technique for identifying interaction partners for an affinity tagged bait protein. The approach relies on the fusion of dual tags to the bait before separate rounds of affinity purification and precipitation. Frequently two specific elution steps are also performed to increase the specificity of the overall technique. In the method detailed here, the two tags used are protein G and a short streptavidin binding peptide; however, many variations can be employed. In our example the tags are separated by a cleavable tobacco etch virus protease target sequence, allowing for specific elution after the first round of affinity purification. Proteins isolated after the final elution step in this process are concentrated before being identified by mass spectrometry. The use of dual affinity tags and specific elution in this technique dramatically increases both the specificity and stringency of the pull-downs, ensuring a low level of background nonspecific interactions.

Shortland A, Chettle J, Archer J, Wood K, Bailey D, Goodfellow I, Blacklaws B A, Heeney J L (2014)

Pathology caused by persistent murine norovirus infection

Journal of General Virology 95 (2), 413-422
Publisher’s version: http://dx.doi.org/10.1099/vir.0.059188-0

Abstract

Subclinical infection of murine norovirus (MNV) was detected in a mixed breeding group of WT and Stat1(-/-) mice with no outward evidence of morbidity or mortality. Investigations revealed the presence of an attenuated MNV variant that did not cause cytopathic effects in RAW264.7 cells or death in Stat1(-/-) mice. Histopathological analysis of tissues from WT, heterozygous and Stat1(-/-) mice revealed a surprising spectrum of lesions. An infectious molecular clone was derived directly from faeces (MNV-O7) and the sequence analysis confirmed it was a member of norovirus genogroup V. Experimental infection with MNV-O7 induced a subclinical infection with no weight loss in Stat1(-/-) or WT mice, and recapitulated the clinical and pathological picture of the naturally infected colony. Unexpectedly, by day 54 post-infection, 50 % of Stat1(-/-) mice had cleared MNV-O7. In contrast, all WT mice remained infected persistently. Most significantly, this was associated with liver lesions in all the subclinically infected WT mice. These data confirmed that long-term persistence in WT mice is established with specific variants of MNV and that despite a subclinical presentation, active foci of acute inflammation persist within the liver. The data also showed that STAT1-dependent responses are not required to protect mice from lethal infection with all strains of MNV.

Adams M J, Lefkowitz E J, King A M Q, Carstens E B (2014)

Ratification vote on taxonomic proposals to the International Committee on Taxonomy of Viruses (2014)

Archives of Virology 159 (10), 2831-2841
Publisher’s version: http://dx.doi.org/10.1007/s00705-014-2114-3

Abstract

Changes to virus taxonomy approved by a vote of all ICTV members in February-March 2014 are reported.
Hocking P M, Stevenson E, Beard P M (2014)

Supplementary biotin decreases tibial bone weight, density and strength in riboflavin-deficient starter diets for turkey poults

British Poultry Science 54 (6), 801-809
Publisher’s version: http://dx.doi.org/10.1080/00071668.2013.860213

Abstract

1. Growth and skeletal responses to different dietary concentrations of riboflavin and biotin were compared in turkey poults from hatch to 21 d of age. The birds were fed on a turkey starter diet with different concentrations of supplementary riboflavin (0, 20 and 40 mg/kg) and biotin (0, 0.3 and 0.6 mg/kg) in a factorial design. 2. Poults fed on diets with no supplementary riboflavin had poor gait scores, decreased times to sit and higher rates of culling compared to poults fed on the control diet (20 mg biotin and 0.3 mg riboflavin/kg diet). Histologically, riboflavin deficiency was associated with a peripheral neuropathy similar to that described previously in chicks and, unexpectedly, in growth plate abnormalities. 3. Tibiae of poults fed on the control diet were larger, more dense, stronger and stiffer than the diets with no supplementary riboflavin. 4. Increasing supplementary biotin in poults fed on diets with no supplementary riboflavin was associated with a decrease in tibia weight, density, strength and stiffness. 5. The results demonstrated that riboflavin deficiency in fast-growing turkey poults was associated with growth retardation, growth plate disturbance and peripheral nerve dysfunction leading to an inability to walk.
Beard P M, Griffiths S J, Gonzalez O, Haga I R, Pechenick Jowers T, Reynolds D K, Wildenhain J, Tekotte H, Auer M, Tyers M, Ghazal P, Zimmer R, Haas J (2014)

A loss of function analysis of host factors influencing vaccinia virus replication by RNA interference

Plos One 9 (6), e98431
Publisher’s version: https://dx.doi.org/10.1128/jvi.01464-16

Abstract

Vaccinia virus (VACV) is a large, cytoplasmic, double-stranded DNA virus that requires complex interactions with host proteins in order to replicate. To explore these interactions a functional high throughput small interfering RNA (siRNA) screen targeting 6719 druggable cellular genes was undertaken to identify host factors (HF) influencing the replication and spread of an eGFP-tagged VACV. The experimental design incorporated a low multiplicity of infection, thereby enhancing detection of cellular proteins involved in cell-to-cell spread of VACV. The screen revealed 153 pro-and 149 anti-viral HFs that strongly influenced VACV replication. These HFs were investigated further by comparisons with transcriptional profiling data sets and HFs identified in RNAi screens of other viruses. In addition, functional and pathway analysis of the entire screen was carried out to highlight cellular mechanisms involved in VACV replication. This revealed, as anticipated, that many pro-viral HFs are involved in translation of mRNA and, unexpectedly, suggested that a range of proteins involved in cellular transcriptional processes and several DNA repair pathways possess anti-viral activity. Multiple components of the AMPK complex were found to act as pro-viral HFs, while several septins, a group of highly conserved GTP binding proteins with a role in sequestering intracellular bacteria, were identified as strong anti-viral VACV HFs. This screen has identified novel and previously unexplored roles for cellular factors in poxvirus replication. This advancement in our understanding of the VACV life cycle provides a reliable knowledge base for the improvement of poxvirus-based vaccine vectors and development of anti-viral theraputics.

Haga I R, Pechenick Jowers T, Griffiths S J, Haas J, Beard P M (2014)

TRAF2 facilitates vaccinia virus replication by promoting rapid virus entry

Journal of Virology 88 (7), 3664-3677
Publisher’s version: http://dx.doi.org/10.1128/jvi.03013-13

Abstract

Tumor necrosis factor receptor (TNFR)-associated factor 2 (TRAF2) is a pivotal intracellular mediator of signaling pathways downstream of TNFR1 and -2 with known pro-and antiviral effects. We investigated its role in the replication of the prototype poxvirus vaccinia virus (VACV). Loss of TRAF2 expression, either through small interfering RNA treatment of HeLa cells or through genetic knockout in murine embryonic fibroblasts (MEFs), led to significant reductions in VACV growth following low-multiplicity infection. In single-cycle infections, there was delayed production of both early and late VACV proteins as well as accelerated virus-induced alterations to cell morphology, indicating that TRAF2 influences early stages of virus replication. Consistent with an early role, uncoating assays showed normal virus attachment but delayed virus entry in the absence of TRAF2. Although alterations to c-Jun N-terminal kinase (JNK) signaling were apparent in VACV-infected TRAF2(-/-) MEFs, treatment of wild-type cells with a JNK inhibitor did not affect virus entry. Instead, treatment with an inhibitor of endosomal acidification greatly reduced virus entry into TRAF2(-/-) MEFs, suggesting that VACV is reliant on the endosomal route of entry in the absence of TRAF2. Thus, TRAF2 is a proviral factor for VACV that plays a role in promoting efficient viral entry, most likely via the plasma membrane.

Walker D, Beard P M, Sharp C P, Philbey A W (2014)

Canine distemper imported into the UK

Veterinary Record 175 (17), 433
Publisher’s version: http://dx.doi.org/10.1136/vr.g6493

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