An amplicon panel for high-throughput and low-cost genotyping of Pacific oyster.

Maintaining genetic diversity in cultured shellfish can be challenging due to high variance in individual reproductive success, founder effects, and rapid genetic drift, but is important to retain adaptive potential and avoid inbreeding depression. To support broodstock management and selective breeding in cultured Pacific oysters (Crassostrea (Magallana) gigas), we developed an amplicon panel targeting 592 genomic regions and SNP variants with an average of 50 amplicons per chromosome. Target SNPs were selected based on elevated observed heterozygosity or differentiation in Pacific oyster populations in British Columbia, Canada. The use of the panel for parentage applications was evaluated using multiple generations of oysters from a breeding program on Vancouver Island, Canada (n = 181) and families selected for Ostreid herpesvirus-1 resistance from the Molluscan Broodstock Program in Oregon, USA (n = 136). Population characterization was evaluated using wild, naturalized, farmed, or hatchery oysters sampled throughout the Northern Hemisphere (n = 190). Technical replicates showed high genotype concordance (97.5%; n = 68 replicates). Parentage analysis found suspected pedigree and sample handling errors, demonstrating the panel's value for quality control in breeding programs. Suspected null alleles were identified and found to be largely population dependent, suggesting population-specific variation impacting target amplification. Null alleles were identified using existing data without the need for pedigree information, and once they were removed, assignment rates increased to 93.0% and 86.0% of possible assignments in the two breeding program datasets. A pipeline for analyzing the amplicon sequence data from sequencer output, amplitools, is also provided.

Endoscopic Repair of CSF Rhinorrhea: Our Experience at a Tertiary Centre.

Spontaneous or post-traumatic CSF (cerebrospinal-fluid) rhinorrhea occurs when there is a skull-base defect. In our study, we tried endoscopic approach, exclusively as surgical modality. To study feasibility of trans-nasal endoscopic approach for repair of skull-base defects and success rates at each anatomical subsite, with complications. Patients, who underwent endoscopic repair of CSF rhinorrhea between 2016 and 2019 were recruited into study. Details of investigative work-up, aetiology, surgery done, site of leak, number of surgical procedures done, post-operative complications and their management, success rate for each anatomical sub-site, was retrieved retrospectively and analyzed. All patients were initially managed with conservative measures before taking up for surgery. Eighteen-patients (male-11, female-7, mean age-40.3 years) with CSF rhinorrhea {spontaneous-5(27.7%), traumatic-13(62.3%)} were found. Sites of leak were cribriform-plate (CP), fovea-ethmoidalis (FE) and posterior-table of frontal-sinus (FS) in 8(44.4%), 5(27.7%) and 5(27.7%) respectively. Twelve (66.6%) patients had no postoperative complications. No patients with defects in CP had post-operative complication. Two (11.1%) patients with FS defect had meningitis, one (5.5%) patient with FS defect developed pneumocephalus. One (5.5%) patient developed frontal sinusitis at end of 4-months. Two (11.1%) patients, each with defects in FE and FS needed a revision repair on postoperative day 0 and 90-till date, none of the patients have any delayed procedure related complications or recurrences. Endoscopic repair of CSF leaks is the norm of the current day due to its minimally invasive nature. However, endoscopic repair of leaks through the frontal sinus were challenging and was associated with a high rate of complications.

To evaluate the efficacy of topical anti-fungal therapy in postoperative cases of COVID Associated Mucormycosis (CAM): A single-blinded randomized control trial.

PURPOSE: To compare the efficacy of 0.1 % w/w Liposomal Amphotericin-B gel with 10 % w/w Povidone-Iodine and saline nasal douching in preventing revision surgery in patients with CAM. STUDY DESIGN: Multi-arm, parallel randomized control trial. STUDY SETTING: The trial was conducted in the Department of ENT, All India Institute of Medical Sciences (AIIMS) Bhubaneswar. METHODS: Participants: Microbiologically and histologically proven cases of mucormycosis who underwent surgical debridement were included in the study. INTERVENTIONS: Postoperatively, patients were randomized into three groups based on the type of topical intervention received, in the form of Lipid-based Amphotericin B gel, povidone­iodine ointment or saline nasal douching. OUTCOME: Requirement of revision surgery in postoperative cases of CAM. RANDOMIZATION: Participants were allotted to one of the three arms by block randomization. BLINDING: Single-blinded trial. RESULTS: Numbers randomized: 15 participants were randomized to each group. Recruitment: Completed recruiting. Numbers analyzed: 15 participants were analyzed in each group. OUTCOMES: Control arm's risk of revision surgery was 4.50 (95 % CI: 1.16-17.44) times than Lipid-based Amphotericin B gel arm and 1.50 (95 % CI: 0.71-3.16) times that of the Povidone- Iodine arm. The difference was statistically significant (p = 0.02) for Amphotericin but not for Povidone-Iodine. CONCLUSIONS: Topical Amphotericin-B gel application in the postoperative cavity can decrease the need for revision surgery and help in early recovery. TRIAL REGISTRATION: CTRI/2021/10/037257. Clinical Trials Registry of India.

Efficacy of Two-phase Treatment Regimen for Rhino-facial Entomophthoromycosis: Our Experience and Review of Literature.

This paper reviews our institutional experience in the management of rhino-facial entomophthoromycosis, a rare subcutaneous fungal infection. We adopt a fixed two-phase protocol comprising an initial 'intensive phase' (two-drug regimen) followed by an 'eradication phase' (monotherapy). We present five patients treated between May 2013 and May 2020 in our institution who were subjected to the mentioned protocol. All five patients received six months of supersaturated potassium iodide solution and itraconazole, followed by four months of monotherapy with only supersaturated potassium iodide solution. All patients responded well to the treatment and were disease-free on follow-up of a minimum two years post-treatment. We recommend this procedure for all typical cases of entomophthoromycosis and suggest avoiding surgical intervention which may disseminate the fungus and delay response to treatment.

Predisposing factors of rhino-orbital-cerebral mucormycosis in patients with COVID 19 infection.

The predisposing factors of invasive fungal disease in COVID 19 infection are still debatable because of the limited human understanding of the virus with the current literature. In this study, we have tried to correlate the various predisposing factors influencing the clinical profile and treatment outcomes in patients with covid associated mucormycosis (CAM). It is a retrospective analysis of cases of CAM during the second wave of COVID 19 infection, which was managed in the department of Otorhinolaryngology from Dec 1, 2020, to June 10, 2021. The detailed clinical, radiological and management of patients with CAM were collected, recorded, evaluated and correlated with the predisposing factors. Of the total, 46 patients, 44(95.65%) were diabetic and 41 patients had a previous history of steroid intake. When clinical parameters were compared between blood sugar < 200 mg/dl and > 200 mg/dl, the old and newly diagnosed diabetes mellitus in patients with CAM, there was no significant differences in any of the above clinical parameters (p > 0.05), except the hospital stay (p = 0,004). Steroid intake in patients with coexisting DM associated with CAM is considered the most important factor for the development of the CAM. There was are no significant difference in any of the clinical/treatment outcomes in patients with CAM with respect to the initial blood sugar, except for the hospital stay. A large sample size with a long-term follow-up period may be needed for a better understanding of common predisposing factors for the development of CAM.

Topographic specificity within membranes of a single muscle detected in vitro.

Spinal motor pools project to target muscles forming distinct rostrocaudal topographic maps during development and regeneration. To define the mechanisms underlying these neuromuscular maps we studied the preferential outgrowth of embryonic spinal cord neurites on muscle membranes from different axial positions and explored the role of ephrin A ligands. We found all five ephrin As (EphAs) expressed in serratus anterior, gluteus maximus and diaphragm muscles. In the diaphragm, four of the five ephrin As are expressed as a caudal to rostral gradient. When ephrin A function is disrupted in muscle membranes by deletion of glycosyl-phosphatidylinositol anchored ephrin A ligands with phosphatidylinositol-specific phospholipase C enzyme treatment or by blocking of ephrin A ligands with EphA fusion proteins, or by genetic manipulation leading to ephrin A2/A5 mutant mice, the spinal cord neurites loose their preference for the membranes of corresponding axial position; suggesting a significant role for ephrins in topographic choices made by growing motor neurons. To closely approximate topographic choices presented to embryonic neurites in vivo, neurites within the phrenic motor pool were challenged to make outgrowth choices on membranes of their normal target, the diaphragm muscle. We observed that neurites from rostral cervical segments (C1 and C2) prefer to grow on rostral diaphragm membranes; caudal cervical neurites (C6-C8) choose caudal diaphragm membranes; a transition of positional preference occurs at C4 and this ability is lost in ephrin A2/A5 mutant mice. These results demonstrate for the first time topographical outgrowth of axons from within a motor pool onto a single target muscle in vitro.