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The Clinicopathology of Foot and Mouth Disease and Molecular Epidemiology of the Viruses Circulated in Cattle of Pabna District, Bangladesh

M R Nazneen
Department of Pathology, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh -2202, Bangladesh

M S Islam
Department of Pathology, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh -2202, Bangladesh

U K Rima
Department of Medicine, Surgery and Obstetrics Faculty of Veterinary & Animal Science, Hajee Mohammad Danesh Science & Technology University, Dinajpur, Bangladesh

D Biswas
Department of Pathology, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh -2202, Bangladesh

R Afroze
Department of Pathology, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh -2202, Bangladesh

M M Hossain
Department of Pathology, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh -2202, Bangladesh

M A H N A Khan
Department of Pathology, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh -2202, Bangladesh

Abstract/ Executive Summary:

This study investigated outbreaks of Foot and Mouth Disease (FMD) in cattle in Pabna district, Bangladesh, during August-September 2015. Out of 100 cattle, 45 were infected with FMD virus, of which five young and two adult cattle died and the postmortem changes in internal organs were recorded. Oral tissue samples from infected cattle (n = 20) of four Upazillas (Sub-district) of Pabna district were collected for viral RNA extraction and serotype identification using reverse transcriptase polymerase chain reaction (RT-PCR). Gross and histopathological changes in oral, pedal and mammary tissues were typical of FMD. Characteristics lesion of 'tiger heart disease' was seen in heart muscle of young and adult cattle. There was thickening of interlobular septum of lungs and characteristics of interstitial pneumonia. The uniplex and multiplex RT-PCR detected FMD viruses (430 bp) and FMD viral serotypes “O” (402 bp) and “Asia 1” (292 bp). Cattle of Sujanagar, Chatmahar and Isshardi Upazillas of Pabna district were infected with FMD viral serotype “O” and responsible for mortality of three young and two adult cattle. FMD viral serotype “ Asia 1” causing death of two young cattle at Pabna Sadar. The fragment (485 bp) of Vp1 gene of FMD viral serotype “O” sequenced showed mutation in main antigenic sites. The phylogenetic analysis carried out with the Vp1 gene of FMD viral ser otype “O” showed the viruses belonging to ME-SA topotype. The death of young and adult cattle was probably due to cardiac and/ or respiratory failure. The adapted RT-PCR protocol can be used in practice for detecting FMD viruses and its serotyping. Larger samples sizes require investigating to identify existing FMD viral serotypes and topotypes in order to design future preventive strategies.

Keywords: cattle, FMD, FMDV,

Location: Sujanagar, Chatmahar, Isshardi and Sadar Upazilla of Pabna district, Bangladesh

Start Date: 01-08-2015

End Date: 30-09-2015

Program Area: Animal Health and Management

Commodity: Cattle

Objectives:

Investigate into the outbreak of foot and mouth disease in cattle and its morbidity and mortality pattern.

Methodology:

This investigation was carried out in Sujanagar, Chatmahar, Isshardi and Sadar Upazilla of Pabna district, Bangladesh during August and September 2015. A total of 25 cattle from each Upazilla were investigated to know the morbidity and mortality pattern of cattle due to FMD. 20 infected cattle from four Upazillas, five from each were selected to collect oral tissues for FMD viral RNA extraction and detection of viral serotypes. Three infected dead calves and two adult dead cattle were investigated by necropsy and histopathology to assess the lesions. Clinicopathological investigations: Ten cattle in a dairy farm (Sujanagar Upazilla, Pabna) were investigated to assess the infectivity during outbreaks. Rectal temperature was recorded for 7 consecutive days. The oral and pedal lesions and relevant signs were recorded. In acute phase of infection scrapings from oral lesions were collected and snap -frozen for the extraction of viral RNA and RT-PCR detection of viral serotypes. Three infected calves (one year of age) were found dead, and post-mortem and histopathological investigations were done. Representative tissues from the affected organs were preserved in 10% buffered neutral formalin, sectioned and stained using haematoxylin and eosin. Extracted FMD viral RNA: The samples collected from oral lesions (n = 20) of acutely infected cattle were snap - frozen for the extraction of viral RNA. Viral RNA from the infected and control tissues (one representative sample from each farm) were extracted using Viral Nucleic Acid Extraction Kit II as per manufacturer’s instructions. The high quality RNA wa s eluted in 30 µL of nuclease-free water, the purity and concentration of viral RNA (A 260/ A 280 and A 260) were determined by using spectrophotometer. The pure RNA has an approximate A 260/ A 280 ratio above 1.8 in Tris -HCl buffer and proven pure RNA was used in RT-PCR detection of the viruses. RT-PCR detection of FMD viruses: Uniplex and multiplex RT-PCR were used to detect FMD viruses and viral serotypes using published primers. The RT-PCR reaction was carried out in 25 µL contain ing 2x Master mix, 70 to 100 ng RNA and 20 pmoL primers in each with the thermal profile. Five infected samples from each of the farms were tested including known positive RNA (from repository) to detect the FMD viruses and viral serotypes circulating. Amplification of VP1 gene and sequencing: Viral RNA extracted from the oral lesion of infected cattle was used in RT-PCR amplification of the fragment of VP1 gene. The RT-PCR amplicons was gel-purified, and the sequences data were analysed to evaluate the extent of mutation and phylogenetic analysis. Sequence editing and alignment were carried out using the “Lasergene” software package. Sequence and phylogenetic analysis: The cDNA of 1D and VP1 genes (639 bp) of FMD viral serotype O was used in analysis. All positions containing gaps and missing data were eliminated; a total of 485 nucleic acid bases remained in the final dataset for translational and phylogenetic analysis. The sequence information was used in Neighbour-Joining tree with kimura-2 parameter to construct the phylogenetic tree and to know the topotype of the viruses. The robustness of the tree topology was assessed with 1,000 bootstrap replicates as implemented in the program.

Source of Information: The Bangladesh Veterinarian (2016) 33(2) : 39 – 50

Article Link (Online Journal):

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

The fragment of Vp1 gene of FMD viral serotype “O” sequenced showed mutation in main antigenic sites. The phylogenetic analysis carried out with the Vp1 gene of FMD viral serotype “O” showed the viruses belonging to ME-SA topotype. The death of young and adult cattle was probably due to cardiac and/ or respiratory failure. The adapted RT- PCR protocol can be used in practice for detecting FMD viruses and its serotyping. Larger samples sizes require investigating to identify existing FMD viral serotypes and topotypes in order to design future preventive strategies.

Knowledge Management: Journal