Biotechnology

Hereditary hypotrichosis is a rare autosomal recessive condition characterized clinically by alopecia. Three consanguineous kindreds with multiple affected individuals were ascertained from different regions of Pakistan. A novel hypotrichosis locus was mapped to a 5.5 cM region on chromosome 18q21.1. A maximum two-point LOD score of 5.25 was obtained at marker D18S36 (h ¼ 0.0). Three genes each for desmoglein and desmocollin proteins are located in this region. The expression in epidermal desmosomes and their connection to the keratin intermediate filaments make these genes excellent candidates for recessive hypotrichosis.

Autosomal recessive nonsyndromic hearing impairment (ARNSHI) is the most frequent form of prelingual hereditary hearing loss in humans. Between 75 and 80% of all nonsyndromic deafness is inherited in an autosomal recessive pattern. Using linkage analysis, we have mapped a novel gene responsible for this form of nonsyndromic hearing impairment, DFNB65, in a consanguineous family from the Azad Jammu and Kashmir regions, which border Pakistan and India. A maximum multipoint LOD score of 3.3 was obtained at marker D20S840. The three-unit support interval is contained between markers D20S902 and D20S430, while the region of homozygosity is flanked by markers D20S480 and D20S430. The novel locus maps to a 10.5-cM region on chromosome 20q13.2-q13.32 and corresponds to a physical map distance of 4.3 Mb. DFNB65 represents the first ARNSHI locus to map to chromosome 20.

The present investigation included a detailed description of the factors directly associated with PPRV infection in Pakistan. A total of 1,056 suspected serum samples were analyzed for the presence of antibodies to PPRV with no history of vaccination against PPR. The samples were collected from sixty two (62) suspected outbreaks from twenty five (25) major regions of the country. Samples were collected from the animals suffering from diarrhea and showing severe respiratory signs. Competitive enzyme linked immuno-sorbant assay (cELISA) was performed to detect the presence of antibodies in the serum against PPRV. Findings suggested that the overall PPR antibody sero-prevalence recorded in sheep was 54.09% as compared to 44.15% in goats. Geography, species, sex, age and season are the major factors associated with PPRV infection. Among various age groups, the animals showed the higher prevalence (67.48% and 52.28% in sheep and goats, respectively) at >2 years as compared with the other age groups. The area-wise highest sero-prevalence was in Sindh province which was 55.10%. It was highest in the plains of Punjab and Sindh province and also in hilly areas (AJK, northern areas and northern Punjab). These findings may be correlated with variations in the sheep and goats husbandry practices within different geographic regions and the topography of different areas. The study also indicated the scenario of antibodies circulation in the population and proving that PPR is now becoming endemic and is one of the major emerging TAD in Pakistan.

Nucleic acid based differentiation of closely related Pasteurella multocida vaccinal strains was performed. Morphological and biochemical characterization, HS-specific and species-specific PCR analysis of Pasteurella multocida vaccinal strains were demonstrated useful in distinguishing hemorrhagic septicemia-causing type B strains. The PCR assay performed for species specific P. multocida by using primer pair KMT1T7 and KMTISP6 resulted in amplification of all the strains.

The present study reports the distribution of different serotypes of foot-and-mouth disease virus (FMDV) in Pakistan during the period 1952-2007. During this time, a total of 1,543 epithelial samples out of 2,484 tested were found positive for various serotypes of FMDV. Serotype O was found to be the most prevalent (p \ 0.001) followed by serotype Asia-1 and A. Serotype C was detected only in 1954, 1963 and 1995. The disease was found to be more prevalent (p \ 0.0001) in cattle than buffaloes. The geographical distribution of 153 laboratory confirmed FMD outbreaks from 2002 to 2007 and the serotypes of the virus involved has been mapped. Higher number of outbreaks of the disease was noted between the months of January to March during this period, which may be attributed to the livestock movement in the country particularly due to religious festival, Eidul Azha, in which the animals are sacrificed.

The present study was carried out to observe inter-strain variation at molecular level in the two vaccinal types (B:3,4 and B:6) of Pasteurella multocida. The micro-organisms were identified and characterized by using conventional microbiological tests. The results of these tests ...

The aim of this study was to determine a relationship between vaccine potency (amount of PD 50 per dose) and fraction of clinically protected cattle following homologous challenge with infectious foot-andmouth disease (FMD) virus, and to determine the effect of method of fractionation, serotype, type of adjuvant, valency and type of virus culture on the dose-response curve. Data from 297 potency tests of FMD vaccines, comprising 4004 vaccinated cattle, performed at the FMD vaccine production facility in the Netherlands, were used for the present study. A generalised linear mixed effect model was used to analyse the results. Our study showed that the relation between FMD vaccine potency and fraction protected was also affected by the serotype and type of adjuvant. No common level of protection could be assigned to all FMD vaccines with the same amount of PD 50 per dose, this information is essential when designing a new standard FMD vaccines control.

Foot-and-mouth disease (FMD) is a highly contagious disease of cloven-hoofed animals including cattle, pigs, sheep and many wildlife species. It can cause enormous economic losses when incursions occur into countries which are normally disease free. In addition, it has long-term effects within countries where the disease is endemic due to reduced animal productivity and the restrictions on international trade in animal products. The disease is caused by infection with foot-and-mouth disease virus (FMDV), a picornavirus. Seven different serotypes (and numerous variants) of FMDV have been identified. Some serotypes have a restricted geographical distribution, e.g. Asia-1, whereas others, notably serotype O, occur in many different regions. There is no cross-protection between serotypes and sometimes protection conferred by vaccines even of the same serotype can be limited. Thus it is important to characterize the viruses that are circulating if vaccination is being used for disease control. This review describes current methods for the detection and characterization of FMDVs. Sequence information is increasingly being used for identifying the source of outbreaks. In addition such information can be used to understand antigenic change within virus strains. The challenges and opportunities for improving the control of the disease within endemic settings, with a focus on Eurasia, are discussed, including the role of the FAO/EuFMD/OIE Progressive Control Pathway. Better control

Foot-and-mouth disease virus (FMDV) is responsible for one of the most economically important infectious diseases of livestock. The virus spreads very easily and continues to affect many countries (mainly in Africa and Asia). The risks associated with the introduction of FMDV result in major barriers to trade in animals and their products. Seven antigenically distinct forms of the virus are known, called serotypes, but serotype C has not been detected anywhere for many years and may now be extinct. The serotypes have been further divided into topotypes (except for serotype Asia-1 viruses, which comprise a single topotype), genotypes, lineages and sub-lineages, which are usually restricted to specific geographical regions. However, sometimes, trans-regional spread of some strains occurs. Due to the error-prone replication of the RNA genome, the virus continuously evolves and new strains frequently arise (e.g. with modified antigenicity). Using nucleotide sequencing technologies, this...

Phylogenetic studies on foot-and-mouth disease viruses (FMDVs) circulating in the West Eurasian region have largely focused on the genomic sequences encoding the structural proteins that determine the serotype. The present study has compared near complete genome sequences of FMDVs representative of the viruses that circulate in this region. The near complete genome sequences (ca. 7600nt) were generated from multiple overlapping RT-PCR products. These amplicons were from FMDVs belonging to serotypes O, A and Asia-1, including members of the O-PanAsia-II and the A-Iran05 lineages, and of Group-II and Group-VII (Sindh-08) within serotype Asia-1, which are currently predominant and widespread in West Eurasia. These new sequences were analysed together with other sequences obtained from GenBank. Comparison of different regions of the FMDVs genomes revealed evidence for multiple, inter-serotypic, recombination events between FMDVs belonging to the serotypes O, A and Asia-1. It is concluded from the present study that dramatic changes in virus sequences can occur in the field through recombination between different FMDV genomes. These analyses provide information about the ancestry of the serotype O, A and Asia-1 FMDVs that are currently circulating within the West Eurasian region.

Background: Foot-and-mouth disease (FMD) is an infectious and highly contagious disease of cloven-hoofed domestic and wild animals, causing heavy economic losses to the livestock industry. Rapid and reliable diagnosis of the disease is essential for the implementation of effective control measures. This study compared sandwich enzyme-linked immunosorbent assay (S-ELISA) and conventional reverse transcription polymerase chain reaction (RT-PCR) for the diagnosis of FMD. Methods: A total of 60 epithelial samples from suspected cases of FMD were tested using both S-ELISA and RT-PCR assays. The level of agreement between the assays was assessed by calculating the Kappa value. Results: S-ELISA detected 38 (63%) samples positive for FMD virus (FMDV). Being predominant, serotype O was detected in 22 (57.9%) of the total samples tested positive, whereas 9 (23.7%) and 7 (18.4%) samples were found positive for serotypes A and Asia-1, respectively. RT-PCR detected viral genome in 51 (85%) of th...

Identification and characterization of the foot-and-mouth disease virus (FMDV) strains circulating in endemic countries and their dynamics are essential elements of the global FMD control strategy. Characterization of FMDV is usually performed in reference laboratories (RL). However, shipping of FMD samples to RL is a challenge due to the cost and biosafety requirements of transportation, resulting in a lack of knowledge about the strains circulating in some endemic areas. In order to simplify this step and to encourage sample submission to RL, we have previously developed a low-cost protocol for the shipment of FMD samples based on the use of lateral flow devices (LFDs) combined with a simple virus inactivation step using 0.2% citric acid. The present study aimed to evaluate this inactivation protocol in the field. For this purpose, 60 suspected FMD clinical samples were collected in Nigeria, Pakistan, and Turkey, three countries where FMD is endemic. Sample treatment, testing on L...