Detection of Peste des petits ruminants virus and goatpox virus from an outbreak in goats with high mortality in Meghalaya state, India.

AIM: We describe a laboratory investigation carried out to confirm the etiology of the heavy mortality (37 animals died out of total 44, i.e. 84%) in goats in Ri-Bhoi district of Meghalaya, Northeast region of India in December 2015. The clinical signs observed were abortion, diarrhea, high fever (up to 104°F), pox lesion in the skin, and respiratory distress. MATERIALS AND METHODS: The samples comprising whole blood, sera, and pox lesion were collected from the animals (n=7) from an outbreak for the screening of peste des petits ruminants (PPR) and poxviruses. The whole blood and sera were used for screening of PPR virus (PPRV) by sandwich enzyme-linked immunosorbent assay (ELISA) and antibody by competitive ELISA as well as detection of PPRV partial N gene by reverse transcription-polymerase chain reaction (PCR). The skin lesions were used for the detection of poxvirus by PCR. RESULTS: The results showed the presence of PPR antigens (58-80%) in the samples by sandwich ELISA and antibody in all the sera samples ranging from 9% to 41% positivity in competitive ELISA. Four samples were positive for PPRV partial N gene. The skin lesion screened for poxvirus was also found to be positive for I3L gene of goatpox virus. CONCLUSION: We confirm the outbreak of disease in goats with high mortality is a case of mixed infection of PPR and goatpox detected for the first time in Northeast India.

Molecular Characterization and Computational Modelling of New Delhi Metallo-ß-Lactamase-5 from an Escherichia coli Isolate (KOEC3) of Bovine Origin.

Emergence of antimicrobial resistance mediated through New Delhi metallo-ß-lactamases (NDMs) is a serious therapeutic challenge. Till date, 16 different NDMs have been described. In this study, we report the molecular and structural characteristics of NDM-5 isolated from an Escherichia coli isolate (KOEC3) of bovine origin. Using PCR amplification, cloning and sequencing of full blaNDM gene, we identified the NDM type as NDM-5. Cloning of full gene in E. coli DH5α and subsequent assessment of antibiotic susceptibility of the transformed cells indicated possible role of native promoter in expression blaNDM-5. Translated amino acid sequence had two substitutions (Val88Leu and Met154Leu) compared to NDM-1. Theoretically deduced isoelectric pH of NDM-5 was 5.88 and instability index was 36.99, indicating a stable protein. From the amino acids sequence, a 3D model of the protein was computed. Analysis of the protein structure elucidated zinc coordination and also revealed a large binding cleft and flexible nature of the protein, which might be the reason for broad substrate range. Docking experiments revealed plausible binding poses for five carbapenem drugs in the vicinity of metal ions. In conclusion, results provided possible explanation for wide range of antibiotics catalyzed by NDM-5 and likely interaction modes with five carbapenem drugs.

Complications of endonasal endoscopic sinus surgery for management of nasal polyp and chronic sinusitis.

Sinusitis is defined as a condition manifested by inflammation of the mucous membrane of the nose & paranasal sinuses, fluid within this cavities and or underlying bone. Chronic rhinosinusitis is diagnosed by the presence of two or more of the following factors facial congestion/fullness, nasal obstruction, nasal discharge or discolored post nasal drainage, hyposmia/anosmia or one major and two of the following minor factors, headache. Halitosis, fatigue, dental pain, cough, ear pressure, fullness. The use of endoscope during surgery of nose & para nasal sinuses improves visualization enable greater preservation of normal structures and reduces the necessity for wide exposure. This cross-sectional study on 50 cases of chronic rhinosinusitis carried out during a period of 6 months. All the cases of both endonasal endoscopic sinus surgery (EESS) group and conventional group assessed clinically endoscopicaly and by imaging both pre and post-operative period. Twenty five cases were EESS group. Twenty five cases of conventional group. Follow-up frequencies are 1st week, 2nd week, 3rd week, 1st month, 3rd months, 6th months. In both type majority of patients in between 21-40 years, most of the cases were male, majority of patients present with headache (80)%, nasal discharge (70%), nasal obstruction (62%), major indication incase of EESS group were ethmoidal polyp 52% & conventional surgery group were chronic sinusitis (56%). Commonest complications were cheek swelling 10(40%) incase of conventional group, crusting 12(84%) in EESS group. Complete recovery incase of EESS were in 21(84%) cases and conventional surgery group 16(64%). Endonasal endoscopic sinus surgery has provided a safe & efficient method for dealing with different sinonasal diseases.

Detection of New Delhi metallo-beta-lactamase and extended-spectrum beta-lactamase genes in Escherichia coli isolated from mastitic milk samples.

In this study, eight Escherichia coli isolates were obtained from milk samples of dairy cattle suffering from clinical/subclinical mastitis. Isolates were characterized for antimicrobial resistance traits and virulence genes. Results revealed that one isolate was harbouring New Delhi metallo-beta-lactamase gene (blaNDM ). Cloning and sequencing of the PCR amplicon confirmed the identity of the gene (GenBank accession no. KC769583) having 100% homology with blaNDM-5 (GenBank accession no. JN104597.1), and this isolate was susceptible to colistin, chloramphenicol and tetracycline only. Moreover, another isolate carried extended-spectrum beta-lactamase (ESBL) gene - blaCTX-M , and all isolates possessed blaTEM gene. Of the eight isolates, only one isolate was positive for shiga toxin gene (stx2), and none were harbouring stx1 gene. Occurrence of New Delhi metallo-beta-lactamase (blaNDM ) in one E. coli isolate and ESBL genes in other isolates poses a potential threat to human health following possible entry and spread through food chain.