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Classical Swine Fever in the Lao Peoples’ Democratic Republic: Virological, Epidemiological and Clinical studies. Classical swine fever (CSF) is a highly contagious virus infection of swine caused by classical swine fever virus (CSFV). The CSF virus is a member of the genus Pestivirus of the family Flaviviridae. Classical swine fever is believed to be endemic in Lao Peoples’ Democratic Republic (Lao PDR). Infectious diseases, including CSF, are a major constraint to pig production in developing countries such as Lao PDR. The aim of this thesis was to investigate aspects and present data regarding the nature of CSF pertinent to Lao PDR. An introduction to Lao PDR, local pig production and a review of pertinent CSF literature is presented in Chapter 1. Low levels of veterinary infrastructure have exacerbated infectious disease problems in developing countries. Chapter 2 of this thesis described the construction and refurbishment of a project laboratory in Lao PDR for the diagnosis of viral diseases, in particular CSF virus Furthermore, a diagnostic specimen submission system was adapted to the local domestic and economic conditions. Poor diagnostic facilities and lack of disease reporting systems in Lao PDR have allowed diseases to spread largely unchecked due to low levels of recognition. The process of development and assessment of appropriate diagnostic assays to the local conditions is presented and discussed in Chapter 3. ELISA and RT-PCR technologies for CSF virus detection in clinical specimens were implemented. Variations to RT-PCR methodologies were also investigated to determine the most suitable technique for the local situation. Results indicated that the RT-PCR methodology was more sensitive than ELISA for the detection of CSF virus in fresh clinical specimens. Notably, the situation was reversed when decomposed samples were tested. Methodologies for the preservation and detection of CSF virus in samples subjected to local tropical condition were also investigated. The proprietary reagent RNAlater ™ was found to be suitable for the preservation of CSF virus RNA under local conditions. Methodologies for CSF virus serology using the ELISA technique are also described. The majority of the pigs in Lao PDR are raised within village small-holder systems, with indigenous breeds being the most popular. Very little is known about the response of indigenous breed pigs to CSF virus infection. Chapter 4 described the pathogenicity of a Lao strain of CSF virus (Lao/Kham225) in both indigenous (Moo Laat) and imported pig breeds (Large white/Landrace cross-breed). Statistically significant (p = 0.05) differences in the breed-related susceptibility to CSF infection were demonstrated in clinical and haematological responses, and post-mortem pathology. The results demonstrated the course of CSF infection in the Large white/Landrace cross-breed was generally more acute than in the indigenous breed. Investigations into the epidemiology of CSF in Lao PDR are presented in Chapter 5. The distribution of CSF outbreaks during the period of mid-1997 to the end of 1999 was investigated and crude incidence results were calculated. Serological surveillance to determine the serological prevalence of CSF virus antibodies in selected regions of Lao PDR was performed during 1997 to 1999. Structured serological surveillance was performed in Vientiane Municipality, Bhorikhamxai, Khammouane and Savannakhet provinces during the survey period. Passive serological surveillance using samples from eight abattoirs in Lao PDR was also performed. Statistically significant (p = 0.05) intra- and inter-provincial differences were noted in a number of the surveys. The success of CSF vaccination via post-vaccination serology was also assessed. The results of the investigations determined that vaccination to prevent CSF infection was insufficient and post-vaccination responses were variable. Phylogenetic and phylogeographic studies to determine the genetic characteristics of Lao PDR and other regional CSF virus isolates are presented in Chapter 6. The 5’-non-coding region and the E2 gene of CSF viruses were investigated to determine genetic relationships between Lao PDR and regional isolates. Genetic typing of all field virus isolates using phylogenetic analysis techniques indicated that all viruses belonged to genogroup 2. Phylogeographic analysis of field viruses revealed a delineation of sub-genogroup allocation on a geographic basis. Members of the sub-genogroup 2.1 originated in Northern and Central regions of Lao PDR. Conversely, members of the sub-genogroup 2.2 originated in Southern and Central regions of Lao PDR. All Vietnamese viruses examined belonged to sub-genogroup 2.2. Phylogenetic analysis indicated that the Vietnamese viruses were largely distinct from Lao and Thai CSF viruses. With the exception of one virus isolate, all Thai viruses also belonged to sub-genogroup 2.2. With the exception of one Vietnamese vaccine virus, all vaccines examined belonged to genogroup 1. A general discussion of the results presented in all chapters, as well as implications for future research into this field, are presented in Chapter 7.