Molecular characterization of lumpy skin disease virus (LSDV) emerged in Bangladesh reveals unique genetic features compared to contemporary field strains.

BACKGROUND: Lumpy skin disease (LSD) is a contagious viral disease of cattle caused by lumpy skin disease virus (LSDV). LSD has recently spread in Asia following outbreaks in the Middle East and Europe. The disease emerged in Bangladesh in July 2019 in the Chattogram district, then rapidly spread throughout the entire country. We investigated six LSD outbreaks in Bangladesh to record the clinical signs and collect samples for diagnostic confirmation. Furthermore, we performed the molecular characterization of Bangladesh isolates, analyzing the full RPO30 and GPCR genes and the partial EEV glycoprotein gene. RESULTS: Clinical observations revealed common LSD clinical signs in the affected cattle. PCR and real-time PCR, showed the presence of the LSDV genome in samples from all six districts. Phylogenetic analysis and detailed inspection of multiple sequence alignments revealed that Bangladesh isolates differ from common LSDV field isolates encountered in Africa, the Middle East, and Europe, as well as newly emerged LSDV variants in Russia and China. Instead, they were closely related to LSDV KSGP-0240, LSDV NI2490, and LSDV Kenya. CONCLUSIONS: These results show the importance of continuous monitoring and characterization of circulating strains and the need to continually refine the strategies for differentiating vaccine strains from field viruses.

Genomic characterisation of Escherichia coli isolated from poultry at retail through Sink Surveillance in Dhaka, Bangladesh reveals high levels of multi-drug resistance.

The surveillance of antimicrobial resistance (AMR) in commensal Escherichia coli from livestock at slaughter is widely employed to assess the potential for risk to humans. There is currently a limited understanding of AMR in Bangladesh poultry at retail in live bird markets, with studies focussing solely on phenotypic characterisation of resistance. To address this evidence gap we performed antimicrobial susceptibility testing and whole genome sequencing on E. coli obtained from chickens from live bird markets in Dhaka in 2018 (n = 38) and 2020 (n = 45). E. coli were isolated from caeca samples following ISO guidelines and sequenced using short and long read methods. Multidrug resistance was extremely common (n = 77) and there was excellent concordance between AMR phenotype and the presence of corresponding AMR genes or mutations. There was considerable genomic diversity, with 43 different sequence types detected. Public health considerations included the high occurrence of resistance to ciprofloxacin (n = 75) associated with plasmid-residing qnrS or mutations in the gyrA and parC chromosomal genes; and the detection of a tigecycline resistant isolate harbouring tet(X4) on an IncHI1A/B-IncFIA mosaic plasmid. Thirty-nine isolates were resistant to azithromycin and harboured mphA, with a significant increase in the incidence of resistance between 2018 and 2020. Although azithromycin is banned for veterinary use in Bangladesh it remains an important treatment option for humans. Interestingly, mphA confers high-level resistance to azithromycin and erythromycin, and the latter is commonly used on poultry farms in Bangladesh. Seven isolates were colistin resistant and carried mcr1. For two isolates hybrid assemblies revealed that mcr1 resided on a highly conserved IncHI2 plasmid that had 93% nucleotide identity to a plasmid from the published genome of an E. coli isolate of Bangladeshi human origin. Six isolates had resistance to third generation cephalosporins, associated with plasmid-residing bla CTX-M-55, bla CTX-M-65, or bla DHA-1. By employing phenotypic and genomic approaches for AMR surveillance we have provided new insights into the potential for One Health AMR linkages in Bangladesh. Employing similar approaches in human and environmental sectors will help inform the One Health approach to addressing AMR, and generate evidence to support mitigation measures such as improved antimicrobial stewardship.

A further outbreak of classical swine fever in indigenous pigs in Kurigram district, Bangladesh.

Deaths of native scavenging pigs were reported in mid-November 2015 at Nageswari sub-district, Kurigram district of Bangladesh. The investigation for a suspected classical swine fever (CSF) outbreak was accomplished via a joint outbreak investigation team from Department of Livestock Services (DLS) and Food and Agriculture Organization, Emergency Center for Transboundary Animal Disease (FAO-ECTAD), Bangladesh. Out of 592 pigs, 396 were infected and among them 263 died. The attack rate and case fatality rate were 66.9% and 66.4%, respectively. The epidemic curve constructed using the data captured from the CSF outbreak site was nearly bell-shaped, indicating a point source epidemic. The basic reproduction numbers (R0) were estimated to be 1.6 (95% Confidence interval [CI]: 1.5-1.7) and 1.5 (95% CI: 1.3-1.7) based on attack rate and exponential growth rate methods, respectively. Adult pigs showed signs of high fever, staggering gait and depression, whereas piglets either died without any premonitory signs or purulent exudates in the eyes were observed. Post-mortem examination was carried out on a 6-month-old piglet. The necropsy findings included were swollen lymph nodes deep red in colour, and haemorrhages on serous and mucous membranes of the intestinal organs together with button-like ulceration in the intestines. Nasal swabs and tissue samples (spleen, lung and liver) were tested using real-time reverse transcriptase polymerase chain reaction (RT-qPCR) and found to be positive for CSF virus. One-step RT-PCR was used to amplify 1,148 base pair of E2 gene in extracted RNA and was sequenced using standard Sanger's sequencing. Phylogenetic analysis revealed the virus as genotype 2.2 that clustered with CSF virus sequences from Bangladesh and India. This is only the second report of a CSF outbreak in Bangladesh. CSF appears to be an emerging transboundary disease in this country. A special programme for controlling swine diseases is needed since pigs are being reared by marginalized communities of Bangladesh to provide livelihoods and food security.