Publications

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

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Lembo T, Oura C, Parida S, Hoare R, Frost L, Fyumagwa R, Kivaria F, Chubwa C, Kock R, Cleaveland S, Batten C (2013)

Peste des petits ruminants infection among cattle and wildlife in Northern Tanzania

Emerging Infectious Diseases 19 (12), 2037-2040

Abstract

We investigated peste des petits ruminants (PPR) infection in cattle and wildlife in northern Tanzania. No wildlife from protected ecosystems were seropositive. However, cattle from villages where an outbreak had occurred among small ruminants showed high PPR seropositivity, indicating that spillover infection affects cattle. Thus, cattle could be of value for PPR serosurveillance.

Abstract

Understanding the influence of non-susceptible hosts on vector-borne disease transmission is an important epidemiological problem. However, investigation of its impact can be complicated by uncertainty in the location of the hosts. Estimating the risk of transmission of African horse sickness (AHS) in Great Britain (GB), a virus transmitted by Culicoides biting midges, provides an insightful example because: (i) the patterns of risk are expected to be influenced by the presence of non-susceptible vertebrate hosts (cattle and sheep) and (ii) incomplete information on the spatial distribution of horses is available because the GB National Equine Database records owner, rather than horse, locations. Here, we combine land-use data with available horse owner distributions and, using a Bayesian approach, infer a realistic distribution for the location of horses. We estimate the risk of an outbreak of AHS in GB, using the basic reproduction number (R0), and demonstrate that mapping owner addresses as a proxy for horse location significantly underestimates the risk. We clarify the role of non-susceptible vertebrate hosts by showing that the risk of disease in the presence of many hosts (susceptible and non-susceptible) can be ultimately reduced to two fundamental factors: first, the abundance of vectors and how this depends on host density, and, second, the differential feeding preference of vectors among animal species

Abstract

Pigs are thought to play a role in the adaptation of avian influenza (AI) viruses to mammalian hosts. To better understand this mechanism and to identify key mutations two highly pathogenic AI (HPAI) viruses (H5N1 and H7N7) were grown in pig cells, To mimic the pressure of an immune response, these viruses were grown in the presence of antiserum to the homologous virus or porcine IFN-gamma. Mutations were identified in both viruses grown in vitro in the presence and absence of antisera or IFN-gamma and included the PB2 mutations, E627K or 627E,D701N, described previously as requirements for the adaptation of AI viruses to mammalian species. Additional mutations were also identified in PB1, HA, NP and M genes for viruses passaged in the presence of immune pressure. The infectivity of these viruses was then assessed using ex vivo pig bronchi and lung organ cultures. For lung explants, higher levels of virus were detected in organ cultures infected with H5N1 HPAI viruses passaged in pig cell lines regardless of the presence or absence of homologous antisera or IFN-gamma when compared with the wild-type parental viruses. No infection was observed for any of the H7N7 HPAI viruses. These results suggest that the mutations identified in H5N1 HPAI viruses may provide a replication or infection advantage in pigs in vivo and that pigs may continue to play an important role in the ecology of influenza A viruses including those of avian origin.

Abstract

Multiple sclerosis is considered a disease of complex autoimmune etiology, yet there remains a lack of consensus as to specific immune effector mechanisms. Recent analyses of experimental autoimmune encephalomyelitis, the common mouse model of multiple sclerosis, have investigated the relative contribution of Th1 and Th17 CD4 T cell subsets to initial autoimmune central nervous system (CNS) damage. However, inherent in these studies are biases influenced by the adjuvant and toxin needed to break self-tolerance. We investigated spontaneous CNS disease in a clinically relevant, humanized, T cell receptor transgenic mouse model. Mice develop spontaneous, ascending paralysis, allowing unbiased characterization of T cell immunity in an HLA-DR15-restricted T cell repertoire. Analysis of naturally progressing disease shows that IFN gamma(+) cells dominate disease initiation with IL-17(+) cells apparent in affected tissue only once disease is established. Tregs accumulate in the CNS but are ultimately ineffective at halting disease progression. However, ablation of Tregs causes profound acceleration of disease, with uncontrolled infiltration of lymphocytes into the CNS. This synchronous, severe disease allows characterization of the responses that are deregulated in exacerbated disease: the correlation is with increased CNS CD4 and CD8 IFN gamma responses. Recovery of the ablated Treg population halts ongoing disease progression and Tregs extracted from the central nervous system at peak disease are functionally competent to regulate myelin specific T cell responses. Thus, in a clinically relevant mouse model of MS, initial disease is IFN gamma driven and the enhanced central nervous system responses unleashed through Treg ablation comprise IFN gamma cytokine production by CD4 and CD8 cells, but not IL-17 responses.

Abstract

Lumpy skin disease is an economically important disease of cattle that is caused by the lumpy skin disease virus (LSDV), which belongs to the genus Capripoxvirus. It is endemic in Africa and outbreaks have also been reported in the Middle-East. Transmission has mostly been associated with blood-feeding insects but recently, the authors have demonstrated mechanical transmission by Rhipicephalus appendiculatus as well as mechanical/intrastadial and transstadial transmission by Amblyomma hebraeum. Saliva is the medium of transmission of pathogens transmitted by biting arthropods and, simultaneously, it potentiates infection in the vertebrate host. This study aimed to detect LSDV in saliva of A. hebraeum and R. appendiculatus adult ticks fed, as nymphs or as adults, on LSDV-infected animals, thereby also demonstrating transstadial or mechanical/intrastadial passage of the virus in these ticks. Saliva samples were tested for LSDV by real-time PCR and virus isolation. Supernatants obtained from virus isolation were further tested by real-time PCR to confirm that the cytopathic effects observed were due to LSDV. Lumpy skin disease virus was detected, for the first time, in saliva samples of both A. hebraeum and R. appendiculatus ticks. At the same time, mechanical/intrastadial and transstadial passage of the virus was demonstrated and confirmed in R. appendiculatus and A. hebraeum.

Abstract

Bovine caesarean section is a common surgery performed by cattle practitioners yet evidence for justifying many aspects of the surgical procedure is lacking. Between 2001 and 2007, questionnaires were used to gather information on 103 cases of caesarean section performed in one, predominantly dairy, veterinary practice. The results showed that the 14-day cow survival rate was 80.6%, and of those surviving beyond this period, 55.4% carried another calf to term, 27.7% were culled due to infertility and 16.9% were culled due to other reasons. Variables associated with reduced 14-day dam mortality included exteriorising the uterus during surgery (odds ratio [OR] 0.018, 95% confidence interval [CI] 0.0019-0.17, P

Abstract

A randomized control trial on verocytotoxigenic Escherichia coli (VTEC)-infected farms found evidence that: (1) keeping animals in the same group; (2) maintaining dry bedding; (3) preventing direct contact with neighbouring cattle; and (4) maintaining a closed herd, were associated with a reduced risk of infection in youngstock aged 3-18 months. This study evaluated these interventions using a cost-effectiveness framework for UK dairy farms. Keeping animals in the same group was considered to have negligible cost and was feasible for herds containing over 77 dairy cows. Assuming equal efficacy of the remaining interventions, preventing direct contact between neighbouring cattle is most cost-effective with a median annual cost of pound2.76 per cow. This compares to pound4.18 for maintaining dry bedding and pound17.42 for maintaining a closed herd using quarantine procedures. Further model validation and exploration of other potential benefits are required before making policy decisions on VTEC control.

Abstract

The genus Orbivirus is the largest of the genera within the family Reoviridae, containing 22 recognised virus species as well as 15 unclassified ‘orbiviruses’, which could potentially represent further new species. The orbiviruses are transmitted by both ticks and/or haematophagous insect vectors. They have a wide host range that includes domestic and wild ruminants, equines, marsupials, sloths, bats, birds and humans. Low-level serological cross-reactions between different species of orbiviruses and lack of reference strains/antisera for existing Orbivirus species make serological identification of new virus isolates difficult. Recently, whole genome sequence data (WGS) has become an important tool for the detection, classification and epidemiological investigations of different pathogens. This study presents full genome sequence database of all known 22 Orbivirus species (including 5 unclassified viruses). Development of novel sequencing strategies and phylogenetic analysis of the orbiviruses using this database has identified five novel Orbivirus species and has facilitated development of a pan-orbivirus RT-PCR assay that can be used to identify the RNA of any Orbivirus species. These techniques will support Orbivirus discovery with greater accuracy than before and can be used for definitive diagnosis of suspected Orbivirus infection.
Madani T A, Abuelzein el T M, Bell-Sakyi L, Azhar E I, Al-Bar H M, Abu-Araki H, Hassan A M, Masri B E, Ksiazek T G (2013)

Susceptibility of tick cell lines to infection with Alkhumra haemorrhagic fever virus

Transactions of the Royal Society of Tropical Medicine and Hygiene 107 (12), 806-11

Abstract

BACKGROUND: Although Alkhumra haemorrhagic fever virus (AHFV) has been isolated from ticks, epidemiological data suggest that it is transmitted from livestock to humans by direct contact with animals or by mosquito bites, but not by ticks. This study was carried out to assess the ability of the virus to replicate in tick cells in vitro. METHODS: AHFV was inoculated into cell lines derived from the hard ticks Hyalomma anatolicum (HAE/CTVM9) and Rhipicephalus appendiculatus (RAE/CTVM1) and the soft tick Ornithodoros moubata (OME/CTVM24). Inoculated cells were directly examined every week for 4 weeks by real-time reverse transcription PCR and by IFAT using polyclonal antibodies. RESULTS: AHFV RNA was detected in all three inoculated tick cell lines throughout the 4-week observation period at levels up to almost twice that of the inoculum, but none of them exhibited a cytopathic effect. AHFV antigen could be detected in all three cell lines by IFAT. Titration of tick cell culture suspension in LLC-MK2 cells yielded AHFV titres of 10(6.6) 50% tissue culture infective dose (TCID50)/ml for OME/CTVM24 and 10(5.5) TCID50/ml for RAE/CTVM1 cells after 4 weeks of culturing; no viable virus was detected in HAE/CTVM9 cells. CONCLUSION: This is the first description of propagation of AHFV in tick cells.

Abstract

A one-step, real-time reverse transcription-loop-mediated isothermal amplification assay (RT-LAMP) for rapid detection and serotyping of Indian foot-and-mouth disease virus (FMDV) is described. The RT-LAMP assay was found to be 103–105 fold more sensitive in comparison with RT-PCR, with a detection limit ranging from 10?3 to 10?5 TCID50 of virus samples of all three serotypes. The RT-LAMP assay and qRT-PCR could detect 100 percent of clinical samples of three serotypes, whereas the RT-PCR detected 69.7% of type O, 58.1% of type A and 60.0% of Asia 1 samples. The qRT-PCR has the same sensitivity as the RT-LAMP. The assay conditions with absence of cross reactivity within the three serotypes of FMDV and FMDV negative samples were established. The RT-LAMP assay could detect 100% of samples stored in FTA cards. In comparison with the performance of the RT-PCR; the RT-LAMP appears to be more sensitive, rapid and specific, with the potential for use as a point-of-care (POC) test, especially in developing countries. The use of FTA cards for the preservation of RNA samples coupled with the RT-LAMP assay for the identification of serotypes may help in achieving improved FMDV serotype identification both in the field and in the laboratory.

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