RESUMEN
Antibiotic residues in pharmaceutical wastewater pose a significant environmental concern due to their potential role in fostering antimicrobial resistance. South Indian pharmaceutical companies produce a wide range of antibiotics. As a result, the industries that discharge water may include antibiotic residues, which could be harmful to the environment. In this study, a novel, quick, accurate, and sensitive approach for the simultaneous detection of 11 antibiotics was established, and triple quadrupole mass spectrometry, ultra-fast liquid chromatography-tandem mass spectrometry (UFLC-MS/MS), and selective solid-phase extraction (SPE) were used for validation. Utilizing a mixed mode reversed-phase/cation-exchange cartridge (SPE using Strata X, 33 µm), the single-cartridge extraction procedure was performed and validated. Relative standard deviations for most of the antibiotics ranged from 3.5 to 0.56 with recoveries ranging from 57 to 85%. The samples were injected into the UFLC-MS/MS apparatus at a volume of 10 µL for analysis. The auto sampler cooler temperature was kept at 150 °C, while the column temperature was kept at 40 °C. After validation, the technique was determined to be linear in the range of 2.0-1000.0 ng/mL. The retention period for antibiotics was between 1.2 and 1.5 min. Antibiotics transitions for multiple reaction monitoring| were between 235.1/105.9 and 711.5/467.9 m/z. The method of analysis took 2.5 min to run completely. Antibiotic residues were efficiently analyzed using the established analytical approach in pharmaceutical wastewater (influent and effluent), surface, and groundwater. Eleven antibiotics were found in the water samples during examination with concentrations ranging between 2.313 and 95.744 ng/L. The procedure was shown to be much more environmentally friendly than other contemporary methods based on the green analytical procedure index's evaluation of greenness. Blue applicability grade index tool indicated the developed method's practicality in comparison with that of other reported method.
RESUMEN
Sudden death of ducklings was reported in a duck farm located at Tiruvallur district in Tamil Nadu, India. Disease investigation began with post mortem findings of dead birds revealing enlarged pale-pink / pale-yellow liver with multifocal petechiae and ecchymosis. A positive amplification with duck hepatitis A virus specific primers by reverse transcription-polymerase chain reaction (RT-PCR) on the tissue samples collected from dead birds indicated infection by duck hepatitis A virus (DHAV), an avian picornavirus, known to cause acute and high-mortality in ducklings. The virus isolation was successful in 9-days old embryonated chicken eggs, in primary chicken embryo fibroblast (CEF) cells and from experimentally infected ducklings. The embryonic death on day 5 to 7 post inoculation in chicken embryos with signs of cutaneous hemorrhage, edema and greenish yellow liver together with histopathology of embryonic liver and kidney further confirmed DHAV infection. TEM analysis of the infected allantoic fluid and infected CEF cell culture supernatant showed the presence of spherical shaped, non-enveloped virion particles of ~ 20-38 nm diameter, typical for DHAV. Experimental infection of ducklings with RT-PCR positive tissue supernatant caused 40% to 50% mortality with typical petechial hemorrhages on the surface of liver. Further, histopathological analysis and RT-PCR of the inoculated duckling's tissues confirmed the presence of DHAV. Nucleotide sequencing of the 5'UTR region and VP1 region confirmed duck hepatitis A virus genotype 2 (DHAV-2). To the best of our knowledge, this is the first report of laboratory confirmation of DHAV-2 in India. This study warrants the need for the extensive epidemiological surveillance to understand the prevalence of DHAV-2 in India and to take appropriate control measures to curtail the disease spread.
