Virulence gene profile and antimicrobial resistance of Staphylococcus aureus isolated from bovine mastitis in Kashmir, India

The Staphylococcus aureusis often responsible for a number of diseases in humans and animals, and it is considered as a main etiological agent of bovine mastitis. The pathogenicity of S. aureus is due to both its ability to resist antibiotics, and the production of toxins. This study investigated virulence genes, prevalence and antibiogram profile of S. aureus isolated from dairy cows suffering from mastitis in the Kashmir. A total of 70 S. aureus isolates were obtained from 250 mastitic milk samples collected from both organized (47/150) and unorganized (23/100) dairy farms. Five pathogenic factors including clfA, hld, seo, lukM, and coa and one resistance gene mecAgene were checked through PCR. Clumping factor gene (clfA) was found in most of the isolates with a percentage of 81.42 % whereas,hld, seo, lukM, and coa were present in 61.2, 54.28, 70, and 71 percent of isolates, respectively. However, amplification of coagene yielded DNA bands of two different sizes. A high percentage of antimicrobial resistance rates were observed, wherein, Ampicillin showed highest resistance with 85.7 %, followed by Kanamycin, Cefotaxime, Sulphadizine and Streptomicin showing 71.42 %, 54.28 %, 51.48 % and 42 %, respectively. A high frequency of Methicillin resistant S. aureus (MRSA 28.57 %) was observed in these isolates and all methicillin resistant isolates were found to be positive for mecA gene via PCR amplification. These results revealed that mastitis-associated S. aureus among bovines of Kashmir is able to accumulate different virulence factors and resistance to antimicrobials, making the treatment of infections difficult

Isolation, virulence gene profiling with molecular cloning of ibeA gene and antibiogram of Escherichia coli from respiratory tract infections of broiler chicken in Kashmir, India

he present study has determined prevalence, serological diversity, virulence gene profileand in vitro antibiogram of Escherichia coli (E. coli) isolates from respiratory tract of broiler chicken in Kashmir valley along with molecular cloning of ibeA gene, an important zoonotic among invasion-associated genes responsible for neonatal meningitis in humans. A total of 224 broilerchickens with a history of respiratory tract infections, in a flock size of 2692 birds from organised and unorganised poultry farms of Kashmir valley, were screened for E. coli isolation. The prevalence of E. coli in birds with respiratory tract infection was found to be 6.38 per cent. Serogrouping revealed that the E. coli isolates were typeable into 10 serogroups with prevalence rate of 8.8% for serogroup O2, along with few isolates rough or untypeable. The screening of E. coli isolates for virulence gene profile revealed the presence of iss in (89.53%), iucDin (85.46 %), tsh in (51.74 %), cva/cvi in (33.14 %), irp2in (55.23 %), papCin (33.72 %), vat in (31.39 %), astAin (38.95 %), fimCin (93.60 %) and ibeAin (1.16 %) isolates. The antibiogram screening of E. coli isolates revealed majority of isolates to be resistant to tetracycline (94.71%), ampicillin (92.67%) and cephalaxin (79.67%).However, most of the isolates were sensitive to amikacin (74.41%) followed by amoxacillin-sulbactam (68.60%) and gentamicin (67.44%). The study thus, suggests that the E. coli isolates from respiratory tract infection of poultry harbour differential virulence genes along with the genes of zoonotic interest and variable antimicrobial resistance, thus, opening the areas for devising dynamic, emergent and effective treatment line

Correlation between serum ferritin level and liver function tests in thalassemic patients receiving multiple blood transfusions

Background:Multiple blood transfusions are the mainstay of thalassemic patients in order to combat the severe anemia. These frequent blood transfusions result in the excessive iron deposition, leading to multiple injuries to a variety of organs in the body. In response to these injuries, the levels of various enzymes are disturbed. The whole phenomena usually involve the interrelation of one parameter with some other. The present study aimed to estimate the levels of serum ferritin and hepatic enzymes and to find out any possible correlation between them in thalassemic patients receiving multiple blood transfusions.  Methods:A total number of 90 thalassemic patients of both sexes ranging from 10-15 years, receiving multiple blood transfusions were included in the present study. Blood samples from all the patients were withdrawn and analyzed for the values of serum ferritin, hemoglobin and hepatic enzymes (serum alanine transaminase, serum aspartate transaminase, serum alkaline phosphatase). Pearson correlation coefficient was applied to observe correlation between serum ferritin level and hepatic enzymes. A P value of ≤0.05 was considered statistically significant.Results:The overall values of serum ferritin, and hepatic enzymes (serum Alanine Transaminase, serum Aspartate Transaminase, serum Alkaline Phosphatase) were remarkably increased than their normal values. However, hemoglobin level was considerably decreased in thalassemic patients. A weak positive insignificant correlation was observed between serum ferritin with hepatic enzymes and hemoglobin in thalassemic patients.Conclusion:Multiple blood transfusions cause iron overload in the body, which in turn, lead to increased serum ferritin levels in thalassemic patients. High levels of hepatic enzymes are somewhat correlated to serum ferritin concentration. However, the exact reason of elevated levels is still unclear. Further detailed studies should be conducted in order to identify the exact mechanism behind this and to search for the promising correlations of various parameters in thalassemic patients receiving multiple blood transfusions.

Frequency of Subdural and Epidural Hematoma in Brain Injury Via Computed Tomography in Trauma Center of DHQ Teaching Hospital Sargodha

At least 10 million TBIs serious enough to result in death or hospitalization occur annually. The mortality associated with acute subdural hematoma has been reported to range from 36-79%. Epidural hematoma occurs in approximately 2% of patients with head injuries and 5–15% of patients with fatal head injuries. Both can be caused by fall, motor vehicle crashes, assaults, blasts and sports activities. CT is best modality for diagnosis of brain injury. Objective:To measure the frequency of subdural and epidural hematoma in brain injury via computed tomography in trauma center of DHQ Teaching Hospital Sargodha.Methodology:In this descriptive study, among 137 patients of traumatic brain injury (TBIs) were selected with age and gender discrimination by convenient sampling, at Department of Radiology, DHQ Teaching Hospital Sargodha. Single slice Computed Tomography Toshiba asteion machine was used.Results:Out of 137 patients collected, 35 were females and 102 were males who visited emergency department due to brain Injury. It shows 25.5% were females and males were 74.5%.Out of 137 patients, 63.5% were injured with RTA and 35.8% came with the history of fall. 67.2% patients present with loss of consciousness, 67.9% patients with skull fractures and 73% with swelling. Out of 137 patients 85.4% develop SDH and 14.6% develop EDH. Conclusion:In this study we conclude that male develop larger number of brain injuries than females. Most patients with history of RTA had epidural hematoma. Females most likely develop subdural hematoma. Most patients with brain injury later develop subdural hematoma. Keywords: Subdural Hematoma, Epidural Hematoma, Traumatic Brain Injury(TBI), Road Traffic Accident(RTA) DOI: 10.7176/JHMN/71-01 Publication date: February 29th 202

Advances in Nematode Identification: A Journey from Fundamentals to Evolutionary Aspects

Nematodes are non-segmented roundworms evenly distributed with various habitats ranging to approximately every ecological extremity. These are the least studied organisms despite being the most diversified group. Nematodes are the most critical equilibrium-maintaining factors, having implications on the yield and health of plants as well as well-being of animals. However, taxonomic knowledge about nematodes is scarce. As a result of the lack of precise taxonomic features, nematode taxonomy remains uncertain. Morphology-based identification has proved inefficacious in identifying and exploring the diversity of nematodes, as there are insufficient morphological variations. Different molecular and new evolving methodologies have been employed to augment morphology-based approaches and bypass these difficulties with varying effectiveness. These identification techniques vary from molecular-based targeting DNA or protein-based targeting amino acid sequences to methods for image processing. High-throughput approaches such as next-generation sequencing have also been added to this league. These alternative approaches have helped to classify nematodes and enhanced the base for increased diversity and phylogeny of nematodes, thus helping to formulate increasingly more nematode bases for use as model organisms to study different hot topics about human well-being. Here, we discuss all the methods of nematode identification as an essential shift from classical morphometric studies to the most important modern-day and molecular approaches for their identification. Classification varies from DNA/protein-based methods to the use of new emerging methods. However, the priority of the method relies on the quality, quantity, and availability of nematode resources and down-streaming applications. This paper reviews all currently offered methods for the detection of nematodes and known/unknown and cryptic or sibling species, emphasizing modern-day methods and budding molecular techniques

Genomic Determinants and Antimicrobial Resistance Pattern of Clinical Isolates of Extended Spectrum Beta Lactamase (ESBL) Producing Escherichia coli

The growing prevalence of antibiotic-resistant bacteria is a worldwide public health apprehension, and Escherichia coli (E. coli) is one of the most commonly implicated bacterial species. Among E. coli isolates, extended-spectrum β-lactamase (ESBL)-producing strains have been identified as a key contributor to antibiotic resistance. Penicillin’s, cephalosporins, and monobactams are only a few of the β-lactam antibiotics that can be rendered inactive by ESBLs. This investigation’s goals were to determine the prevalence of ESBL-producing E. coli isolates found in clinical samples and to analyze the distribution of the blaTEM, blaSHV, and blaCTX-M genes among them. Additionally, we aimed to determine the antibiotic susceptibility patterns of these isolates to other antibiotics. Clinical isolates from urine, ear swab, and wound/pus swabs were collected from patients with suspected E. coli bacterial infections from different regions of north India viz., SKIMS-JVC medical college and NABL accredited Dr. Qadri’s Lab both from Srinagar Kashmir valley region and SMSR, SU from Greater Noida UP. Standard laboratory techniques were used to identify E. coli isolates, and the combined disc method and other phenotypic confirmation techniques were used to confirm ESBL formation. PCR amplification and sequencing were used to find the blaTEM, blaSHV, and blaCTX-M genes. The Kirby-Bauer disc diffusion method was used to test the antimicrobial susceptibility of various bacteria to different antibiotics. A total of 210 E. coli isolates were collected from different clinical samples and only 158 isolates showed positive results for ESBL by DDST and phenotypic confirmatory tests. Of these, 124 (78.48%) were ESBL-producing isolates. We found that blaTEM was the most prevalent gene (45.16%), followed by blaCTX-M (34.16%) and blaSHV (12.09%). Antimicrobial resistance profiles were assessed for each of the 120 isolates. Ampicillin and Cefepime were the most resistant drugs to ESBL-producing isolates, followed by Gentamicin, Ceftriaxone, and Cefixime

Termodinamička svojstva nove invertaze proizvedene s pomoću dvostruko mutiranoga kvasca Kluyveromyces marxianus

Kinetics of intracellular invertase production employing a double mutant derivative of Kluyveromyces marxianus was optimized by varying different process variables in a 23-litre fermentor. The maximum volumetric rate (QP) and invertase yield (YP/S) by M15 mutant were 1222 U/(L·h) and 160 U/g of substrate utilized, respectively (2-fold more than those of parental strain) at 50 °C on the molasses (150 g/L of total fermentable sugars) at pH=5.5. Glucose or sucrose (100, 150 or 170 g/L) did not repress invertase catabolically under the optimized fermentation conditions, contrary to the previous reports on other yeasts and filamentous fungi, where catabolite repression of sugars was predominant. Invertases derived by the wild (IW) and mutant (IM) strains were purified employing ammonium sulphate precipitation, and then characterized by column chromatographic techniques both kinetically and thermodynamically. The acidic limb of invertases was missing and collation of pKa and the heat of ionization values indicated that carboxyl groups were involved in proton transfer during active catalysis. Ratios of Kcat/Km and vmax/Km indicated that IM was significantly more specific for sucrose hydrolysis. The IM exhibited stability in different buffers at pH=3.0–10.0 and temperature of 50–70 °C, as reflected by long half-lives. IM showed significantly lower values of enthalpy of activation (ΔH*) and entropy of activation (ΔS*), while Gibbs free energy (ΔG*) was significantly increased at higher temperatures, making the IM thermodynamically more thermostable. Thus IM could be used as a catabolite-resistant invertase for the production of fructose syrup or high gravity ethanol.Promjenom različitih varijabla procesa optimirana je kinetika proizvodnje intracelularne invertaze u 23-litarskom fermentoru s pomoću dvostruko mutiranoga kvasca Kluyveromyces marxianus. Maksimalni protok (QP) i prinos invertaze, dobivene s pomoću mutanta M15 na podlozi od melase (150 g/L ukupnih fermentirajućih šećera), pri 50 °C i pH=5,5 bili su: 1222 U/(L•h) i 160 U/g podloge, tj. dvostruko veći nego s roditeljskim sojem. Za razliku od prijašnjih istraživanja na drugim kvascima i filamentoznim plijesnima, u kojima su šećeri uzrokovali kataboličku represiju enzima, u ovom ispitivanju glukoza ili saharoza (u koncentracijama od 100, 150 ili 170 g/L) nisu potaknule kataboličku represiju invertaze pri optimiranim uvjetima fermentacije. Invertaze dobivene s pomoću divljeg i mutiranog soja kvasca pročišćene su taloženjem pomoću amonijeva sulfata i razdvojene kromatografijom na stupcu, te su ispitana njihova kinetička i termodinamička svojstva. Dobivene invertaze nisu imale izraženi pad aktivnosti u kiselom području, a usporedbom vrijednosti pKa i energije ionizacije utvrđeno je da su karboksilne skupine enzima sudjelovale u prijenosu protona tijekom aktivne katalize. Iz omjera Kcat/Km i νmax/Km vidljivo je da je invertaza dobivena s pomoću mutiranog soja kvasca specifična za hidrolizu sukroze, te da je stabilna u različitim puferima pH-vrijednosti od 3 do 10 i pri temperaturi od 50 do 70 °C, što se vidi iz dugog poluvremena njezina raspada. Ta je invertaza imala kudikamo niže vrijednosti entalpije (ΔH*) i entropije (ΔS*), termodinamički je bila stabilnija, a to se vidi iz porasta Gibbsove slobodne energije pri višim temperaturama. Stoga je zaključeno da je invertaza dobivena s pomoću mutiranog soja kvasca Kluyveromyces marxianus otporna na kataboličku represiju te se može upotrijebiti u proizvodnji fruktoznoga sirupa ili etanola s velikim udjelom alkohola

SGCD Homozygous Nonsense Mutation (p.Arg97∗) Causing Limb-Girdle Muscular Dystrophy Type 2F (LGMD2F) in a Consanguineous Family, a Case Report

Background: Limb-girdle muscular dystrophy (LGMD) is an increasingly heterogeneous category of inherited muscle diseases, mainly affecting the muscles of shoulder areas and the hip, segregating in both autosomal recessive and dominant manner. To-date, thirty-one loci have been identified for LGMD including seven autosomal dominant (LGMD type 1) and twenty four autosomal recessive (LGMD type 2) inherited loci.Methodology/Laboratory Examination: The present report describes a consanguineous family segregating LGMD2F in an autosomal recessive pattern. The affected individual is an 11-year-old boy having two brothers and a sister. Direct targeted next generation sequencing was performed for the single affected individual (VI-1) followed by Sanger sequencing.Results: Targeted next generation sequencing revealed a novel homozygous nonsense mutation (c.289C>T; p.Arg97∗) in the exon 3 of the delta-sarcoglycan (SGCD) gene, that introduces a premature stop codon (TCA), resulting in a nonsense mediated decay or a truncated protein product.Discussion and Conclusion: This is the first report of LGMD2F caused by an SGCD variant in a Pakistani population. The mutation identified in the present investigation extends the body of evidence implicating the gene SGCD in causing LGMD2F and might help in genetic counseling, which is more important to deliver the risk of carrier or affected in the future pregnancies

Burnout among surgeons before and during the SARS-CoV-2 pandemic: an international survey

Background: SARS-CoV-2 pandemic has had many significant impacts within the surgical realm, and surgeons have been obligated to reconsider almost every aspect of daily clinical practice. Methods: This is a cross-sectional study reported in compliance with the CHERRIES guidelines and conducted through an online platform from June 14th to July 15th, 2020. The primary outcome was the burden of burnout during the pandemic indicated by the validated Shirom-Melamed Burnout Measure. Results: Nine hundred fifty-four surgeons completed the survey. The median length of practice was 10 years; 78.2% included were male with a median age of 37 years old, 39.5% were consultants, 68.9% were general surgeons, and 55.7% were affiliated with an academic institution. Overall, there was a significant increase in the mean burnout score during the pandemic; longer years of practice and older age were significantly associated with less burnout. There were significant reductions in the median number of outpatient visits, operated cases, on-call hours, emergency visits, and research work, so, 48.2% of respondents felt that the training resources were insufficient. The majority (81.3%) of respondents reported that their hospitals were included in the management of COVID-19, 66.5% felt their roles had been minimized; 41% were asked to assist in non-surgical medical practices, and 37.6% of respondents were included in COVID-19 management. Conclusions: There was a significant burnout among trainees. Almost all aspects of clinical and research activities were affected with a significant reduction in the volume of research, outpatient clinic visits, surgical procedures, on-call hours, and emergency cases hindering the training. Trial registration: The study was registered on clicaltrials.gov "NCT04433286" on 16/06/2020