Isolation and adaptation of bovine herpes virus Type 1 in embryonated chicken eggs and in Madin–Darby bovine kidney cell line

Aim: Objective of the present study was to isolate bovine herpes virus Type 1 (BHV-1) from semen of infected bull and to adapt it onto embryonated eggs and Madin–Darby bovine kidney (MDBK) cell line. Further, the virus was identified by agar gel immunodiffusion (AGID) test. Materials and Methods: Semen samples were collected from five BHV-1 positive bulls previously confirmed for the presence of antibodies against BHV-1 using avidin-biotin enzyme linked immunosorbent assay test. The virus from semen samples was adapted in chorioallantoic membrane (CAM) of 11-day-old embryonated chickens eggs and in MDBK cell line. The presence of BHV-1 in infected CAM and cell culture fluid was confirmed by AGID test. Results: Virus infected CAM showed edema, congestion and thickening at first passage level. Small foci ranged from 1 to 2 mm in diameter, scattered all over the membrane were observed at first passage. More severe changes were observed in CAM after serial passaging. The large pock lesions, round in shape with opaque raised edge and depressed gray central area of necrosis ranged from 3 to 5 mm in diameter were developed at fourth passage. Blind passages in MDBK cell culture were made. The MDBK cell line at second passage level showed characteristic cytopathic effect viz. rounding of cells with shrinkage, followed by aggregation or clumping of cells which progressed rapidly and appeared as “bunch of grapes” at 72 h post inoculation. Few cells become elongated when compared with uninfected controls. A homogenate of CAM with distinct pock lesions and infected cell culture fluid developed precipitation line within 48 h against specific anti-BHV-1 immune serum by AGID test. Conclusion: BHV-1 was easily adapted in CAM of chicken embryos and in MDBK cell line. Virus infected CAM and cell culture fluid showed precipitin band by AGID test

Effect of angiotensin-converting enzyme inhibitor and angiotensin receptor blocker initiation on organ support-free days in patients hospitalized with COVID-19

IMPORTANCE Overactivation of the renin-angiotensin system (RAS) may contribute to poor clinical outcomes in patients with COVID-19. Objective To determine whether angiotensin-converting enzyme (ACE) inhibitor or angiotensin receptor blocker (ARB) initiation improves outcomes in patients hospitalized for COVID-19. DESIGN, SETTING, AND PARTICIPANTS In an ongoing, adaptive platform randomized clinical trial, 721 critically ill and 58 non–critically ill hospitalized adults were randomized to receive an RAS inhibitor or control between March 16, 2021, and February 25, 2022, at 69 sites in 7 countries (final follow-up on June 1, 2022). INTERVENTIONS Patients were randomized to receive open-label initiation of an ACE inhibitor (n = 257), ARB (n = 248), ARB in combination with DMX-200 (a chemokine receptor-2 inhibitor; n = 10), or no RAS inhibitor (control; n = 264) for up to 10 days. MAIN OUTCOMES AND MEASURES The primary outcome was organ support–free days, a composite of hospital survival and days alive without cardiovascular or respiratory organ support through 21 days. The primary analysis was a bayesian cumulative logistic model. Odds ratios (ORs) greater than 1 represent improved outcomes. RESULTS On February 25, 2022, enrollment was discontinued due to safety concerns. Among 679 critically ill patients with available primary outcome data, the median age was 56 years and 239 participants (35.2%) were women. Median (IQR) organ support–free days among critically ill patients was 10 (–1 to 16) in the ACE inhibitor group (n = 231), 8 (–1 to 17) in the ARB group (n = 217), and 12 (0 to 17) in the control group (n = 231) (median adjusted odds ratios of 0.77 [95% bayesian credible interval, 0.58-1.06] for improvement for ACE inhibitor and 0.76 [95% credible interval, 0.56-1.05] for ARB compared with control). The posterior probabilities that ACE inhibitors and ARBs worsened organ support–free days compared with control were 94.9% and 95.4%, respectively. Hospital survival occurred in 166 of 231 critically ill participants (71.9%) in the ACE inhibitor group, 152 of 217 (70.0%) in the ARB group, and 182 of 231 (78.8%) in the control group (posterior probabilities that ACE inhibitor and ARB worsened hospital survival compared with control were 95.3% and 98.1%, respectively). CONCLUSIONS AND RELEVANCE In this trial, among critically ill adults with COVID-19, initiation of an ACE inhibitor or ARB did not improve, and likely worsened, clinical outcomes. TRIAL REGISTRATION ClinicalTrials.gov Identifier: NCT0273570

Occurrence and characteristics of extended-spectrum β-lactamases producing Escherichia coli in foods of animal origin and human clinical samples in Chhattisgarh, India

Aim: To assess the prevalence of antimicrobial resistance producing extended-spectrum β-lactamases (ESBL) (blaTEM, blaSHV, and blaCTX-M) genes in Escherichia coli isolated from chicken meat, chevon meat, raw milk, and human urine and stool samples collected from tribal districts of Chhattisgarh, viz., Jagdalpur, Dantewada, Kondagaon, and Kanker. Materials and Methods: A total of 330 samples, comprising 98 chicken meat, 82 chevon meat, 90 raw milk, and 60 human urine and stool samples, were processed for isolation of E. coli. Isolates were confirmed biochemically and further tested against commonly used antibiotics to know their resistant pattern. The resistant isolates were tested for ESBL production by phenotypic method followed by characterization with molecular method using multiplex-polymerase chain reaction technique. Results: Overall 57.87% (191/330) samples were found positive for E. coli, which include 66.32% (65/98) chicken meat, 46.34% (38/82) chevon meat, 81.11% (73/90) raw milk, and 25% (15/60) human urine and stool samples. Isolates showed the highest resistance against cefotaxime (41.36%) followed by oxytetracycline (34.03%), ampicillin (29.31%), cephalexin (24.60%), cefixime (16.75%), and ceftazidime (13.08%). Phenotypic method detected 10.99% (21/191) isolates as presumptive ESBL producers, however, molecular method detected 3.66% (7/191), 2.09% (4/191), and 0.00% (0/191) prevalence of blaTEM, blaCTX-M, and blaSHV, respectively. Conclusion: The present study indicates a high prevalence of E. coli in raw chicken meat, chevon meat, and milk due to poor hygienic practices. The antibiotic susceptibility test detected the presence of the resistance pattern against ESBL in E. coli isolated from raw chicken meat, chevon meat, milk, and also in human clinical samples is of great concern. The appearance of E. coli in the human food chain is alarming and requires adaptation of hygienic practices and stipulate use of antibiotics

A Comparative Study Between Two Different Doses of Dexmedetomidine Combined with Ropivacaine in Ultrasound Guided Supraclavicular Brachial Plexus Block

Background: Though, various studies have been conducted to show the efficacy of            dexmedetomidine as an adjuvant to brachial plexus block, there is no clear consensus for its optimal dose. We compared 2 different doses of dexmedetomidine (1mcg/kg Vs 2mcg/kg) with ropivaciane in USG guided brachial plexus block for the quality of anesthesia and analgesia. Materials and Methods: Fifty patients (18-50years, 50-60kgs), ASA PS I and II undergoing surgery of upper limb were enrolled in this prospective, double blind, randomized control trial. Group Rd received 19ml of 0.5% Ropivaciane with 1mcg/kg  of dexmedetomidine (total 20ml), and Group RD received 19ml of 0.5% Ropivacaine with 2mcg/kg of dexmedetomidine (total 20ml). Onset and duration of sensory and motor block, duration of analgesia, sedation score, hemodyanamic changes were compared. We also monitored for various un towards effects. Results: The onset time of sensory and motor block (9.36±1.114 mins and 14.40 ± 1.528 mins in group Rd vs 8.32 ±0.945 mins and 12.40 ± 1.21 mins in group RD) were not significantly different between two groups (p-value &gt; 0.05).The duration of sensory and motor block (596.20 ± 76.859 mins and 541.20±81.564 mins in group Rd vs 730.80 + 65.187 mins and 659.80±93.607 mins in group RD) were not significantly longer in group RD than group Rd (p-value &gt; 0.05). The duration of analgesia (626.00±70.475 mins in group Rd vs 754.00±60.139 mins in group RD) were not significantly longer in group RD than group Rd (p-value &gt; 0.05). Hemodynamic parameters were also comparable between two groups (p-value &gt;0.05) but 3 patients in RD group (2mcg/kg) developed bradycardia and required treatment with atropine whereas in Rd group (1mcg/kg) none developed bradycardia. Conclusions: There is no significant difference in the onset and duration of block between the two groups. However, higher dose of    dexmedetomidine is associated bradycardia. Hence, in comparision with 2mcg/kg, 1mcg/kg of   dexmedetomidine is better adjuvant to 0.5% ropivaciane in terms of safety and effectiveness.Keywords: dexmedetomidine; ropivacaine; supraclavicular brachial plexus block. </p

Development of Monsoonal Rainfall Intensity-Duration-Frequency (IDF) Relationship and Empirical Model for Data-Scarce Situations: The Case of the Central-Western Hills (Panchase Region) of Nepal

Intense monsoonal rain is one of the major triggering factors of floods and mass movements in Nepal that needs to be better understood in order to reduce human and economic losses and improve infrastructure planning and design. This phenomena is better understood through intensity-duration-frequency (IDF) relationships, which is a statistical method derived from historical rainfall data. In Nepal, the use of IDF for disaster management and project design is very limited. This study explored the rainfall variability and possibility to establish IDF relationships in data-scarce situations, such as in the Central-Western hills of Nepal, one of the highest rainfall zones of the country (~4500 mm annually), which was chosen for this study. Homogeneous daily rainfall series of 8 stations, available from the government's meteorological department, were analyzed by grouping them into hydrological years. The monsoonal daily rainfall was disaggregated to hourly synthetic series in a stochastic environment. Utilizing the historical statistical characteristics of rainfall, a disaggregation model was parameterized and implemented in HyetosMinute, software that disaggregates daily rainfall to finer time resolution. With the help of recorded daily and disaggregated hourly rainfall, reference IDF scenarios were developed adopting the Gumbel frequency factor. A mathematical model [i = a(T)/b(d)] was parameterized to model the station-specific IDF utilizing the best-fitted probability distribution function (PDF) and evaluated utilizing the reference IDF. The test statistics revealed optimal adjustment of empirical IDF parameters, required for a better statistical fit of the data. The model was calibrated, adjusting the parameters by minimizing standard error of prediction; accordingly a station-specific empirical IDF model was developed. To regionalize the IDF for ungauged locations, regional frequency analysis (RFA) based on L-moments was implemented. The heterogeneous region was divided into two homogeneous sub-regions; accordingly, regional L-moment ratios and growth curves were evaluated. Utilizing the reasonably acceptable distribution function, the regional growth curve was developed. Together with the hourly mean (extreme) precipitation and other dynamic parameters, regional empirical IDF models were developed. The adopted approach to derive station-specific and regional empirical IDF models was statistically significant and useful for obtaining extreme rainfall intensities at the given station and ungauged locations. The analysis revealed that the region contains two distinct meteorological sub-regions highly variable in rain volume and intensity

Characterization of Salmonella Gallinarum from an outbreak in Raigarh, Chhattisgarh

Aim: The present investigation was conducted to isolate and characterize Salmonella Gallinarum from an outbreak of fowl typhoid in layer birds. Materials and Methods: Clinically ill and dead layer birds from an outbreak were investigated. History, clinical signs, and postmortem lesions were suggestive of fowl typhoid. Postmortem samples including heart blood, intestinal contents, pieces of ovary, and liver were collected and processed immediately for bacterial culture, serotyping and antibiotic sensitivity tests. Isolates were further screened for the presence of extended spectrum beta lactamase (ESBL) (blaTEM) gene by polymerase chain reaction. Results: On the basis of cultural, staining and biochemical characteristics; three bacterial isolates were confirmed as S. Gallinarum. On serotyping, somatic antigen O: 9 and 12 with nonflagellated antigen were detected in all three isolates. Isolates were intermediate sensitive to amoxycillin, amoxyclav, gentamicin and ciprofloxacin and resistant to most of the antibiotics including chloramphenicol, ampicillin, ceftazidime, cefexime, cefepime, azithromycin, nalidixin, tetracycline, oxytetracycline, and streptomycin. Two isolates were found to harbor ESBL (blaTEM) gene. Conclusion: Beta lactamase producer S. Gallinarum was confirmed as cause of increased mortality in layer birds during present investigation. Existence of multi drug resistant Salmonella poses serious threat to poultry industry in Chhattisgarh

Adverse events following Immunization with Sinopharm (Vero Cell) inactivated COVID-19 vaccine: Adverse events of Vero Cell vaccine

Introduction: Various types of COVID 19 vaccines are being used globally to control the current pandemic. Post-licensure surveillance of vaccines is essential to ensure safety. This study aimed to determine Adverse Events Following Immunization (AEFIs) of Sinopharm (Vero cell), the inactivated COVID-19 vaccine from China.   Method: This is a cross-sectional observational study conducted at Patan hospital, Patan Academy of Health Sciences (PAHS). Vaccine recipients between April and May were contacted through a phone call after 72 h of vaccination to record the AEFIs. Pattern and distribution of AEFIs were analyzed. Ethical approval was taken from PAHS IRC.   Result: A total of 6142 individuals got the first dose of the vaccine and out of them we were able to contact 4574 through phone calls. Of the 4574, only 941 were included for the follow-up phone call after the second dose of the vaccine. A total of 1336 AEFIs were reported in 868(19%) first dose vaccine recipients while 147 AFEIs were reported in 105(11.2%) second dose vaccine recipients.  The frequently reported AEFIs were pain at the injection site, lethargy, headache, muscle ache, and fever. All the AEFIs were mild to moderate in severity. Most of the AEFIs started within 24 h and resolved within 72 h.   Conclusion: The Sinopharm (Vero cell) vaccine was found to have mild to moderate AEFIs in our study cohort and no case of severe AEFI was identified.