Antibiotic-resistant bacteria in bovine milk in India.

Antibiotic resistance (ABR) is a global issue that draws the attention of all healthcare experts in the veterinary and medical fields. Of various factors, indiscriminate and unregulated antibiotic usage in the animals reared for food production, especially in cows and buffaloes suffering from mastitis, contribute significantly to the rising incidence of resistant bacteria. A literature survey reveals the spread of resistant strains of mastitis-causing bacteria, like Staphylococcus aureus and Escherichia coli, to humans. In addition, antibiotic residues detected in milk samples against all major groups of antibiotics are likely to enter the human body through the food chain and aggravate the condition. The cumulative effects of ABR have emerged as a silent killer. The benefits of systematic surveillance on ABR in India are yet to be available. Here is an attempt to understand the ABR burden in India associated with bovine milk and its mitigation strategies.

Demographic profile and clinical characteristics of Fuchs' endothelial corneal dystrophy in a tertiary eye care center.

Purpose: This study was performed to determine the demographic profile and clinical characteristics in patients with Fuchs' endothelial corneal dystrophy (FECD) reporting to a tertiary eye care center in India. It is a retrospective, single-center, observational study. Methods: The study included 280 patients (559 eyes) diagnosed with FECD presenting between January 2013 and December 2020. The data was collected from the electronic medical record system of the institute. Patient data included demographic features, clinical characteristics, investigations, and surgical interventions. Results: The mean age of the patients was 62 years. Late-onset FECD (95.7%) was more common than early-onset FECD (4.3%). Male: female ratio for late-onset FECD and early-onset FECD was 1:1.65 and 3:1, respectively. More than one-third of the patients had associated systemic history. Preexisting ocular diseases were seen in 5.9% of eyes. Blurring of vision was seen in 383 eyes (68.5%), 13 eyes (2.1%) had glare, and 163 eyes (29.2%) were asymptomatic. A total of 113 surgical interventions were done in 108 eyes (including repeat transplants). Only cataract surgery was done in 40 (7.2%) eyes, whereas penetrating keratoplasty, Descemet stripping endothelial keratoplasty, and Descemet membrane endothelial keratoplasty without or with cataract surgery (sequential or triple procedure) were done in 12 (2.1%), 47 (8.4%), and 14 (2.5%) eyes, respectively. Conclusion: Patients with FECD present mostly during the sixth decade. Posterior lamellar keratoplasty is the most common transplant procedure being performed on FECD patients.

Comparative evaluation of diagnostic methods for detection of Theileria spp. in cows.

Theileriosis is a hemoprotozoan illness of cattle in tropical regions that poses a severe economic loss to dairy farmers in the form of production loss and mortality. We designed and optimized a multiplex real-time PCR by using Taq-Man® probe for detection and quantification of Theileria orientalis and Theileria annulata simultaneously by targeting 18 s rRna and MPSP (surface merozoite protein) genes, respectively. Fifty-five EDTA blood samples from clinically Theileria-suspected cows of three Theileria-endemic districts of Odisha were processed using acridine dye based fluorescent microscopy, Giemsa staining, and PCR. PCR revealed T. annulata and T. orientalis in 11/42 (26.11%) and 24/42 (57.14%) cases, respectively. Mixed infection due to both the Theileria spp. was recorded in 7/42 (16.66%). On comparison with gold standard test (PCR), the accuracy, sensitivity, and specificity were 92.72, 95.12, and 85.71% for Giemsa staining and 96.36, 97.56, and 92.85% for acridine orange dye. Multiplex real time PCR using Taq-Man probe detected two species of T. annulata and T. orientalis simultaneously. Acridine dye based fluorescent microscopy is comparatively easy and rapid method in detection of Thelieria spp.

Multidrug-resistant keratitis: challenging yet manageable.

PURPOSE: To study the incidence, clinical features and outcomes of multidrug-resistant (MDR) bacterial keratitis. METHODS: All cases of MDR-bacterial keratitis presenting to our institute over a period of 2 years were retrospectively analysed. Details of risk factors, size and depth of infiltrate, treatment, and outcome were noted. Antibiotic susceptibility tests were done on the ocular isolates from the culture of samples obtained from ocular infections, and resistance or sensitivity of the organisms to the commonly used antibiotics was studied. RESULTS: Forty patients were diagnosed with MDR-bacterial keratitis in the study period. The mean age of patients was 50.9±25.4 years. Most common risk factors were vegetative trauma (n=12, 30.0%), followed by corneal transplantation (n=7, 17.5%) and systemic comorbidities (n=7, 17.5%). Infiltrate was small (<6 mm) in 22 (55%) and large (>6 mm) in 18 (45%) patients. It involved the superficial, mid and deep stroma in 11 (27.5%), 9 (22.5%) and 15 (37.5%) cases, respectively. Gram-negative bacilli (n=18, 45%) were the maximum, among which Pseudomonas aeruginosa (15%) was the most common. Resistance to 3 (n=17, 42.5%) and 4 (n=17, 42.5%) classes of antibiotics was the most commonly observed. One (2.5%) patient showed resistance to all seven classes of drugs tested. Complete resolution of infection was seen in 15 (37.5%) MDR patients on medical management alone. Five (12.5%) patients underwent therapeutic penetrating keratoplasty. Size of the infiltrate was found to have a significant correlation with the outcome (p=0.002). CONCLUSION: MDR keratitis, despite being a challenge to treat, can be successfully managed by medical therapy alone, if appropriate therapy is started early in the clinical course.

Flavonoids for the Treatment of Breast Cancer, Present Status and Future Prospective.

Breast Cancer is one of the most notorious cancer affecting women globally. Current therapies available for breast cancer treatment have certain limited efficacy; develop drug resistance and severe adverse effects. Thus, identifying novel therapies for treatment will reduce the devastating effect on cancer survivors. The exhilarating and fastgrowing studies on flavonoids have evidenced that it has the potential to inflect various antitumor activity and modulate various signal transduction pathways in carcinogenesis. Flavonoids also have been found to regulate cellular metabolism and oxidative stress, cell cycle progression, angiogenesis and metastasis, ultimately preventing the progression of the diseases. As per the reports, a flavonoid-rich diet appears to be the most potent and promising approach to abate the risk of cancer. Thus, now a day, these are the prime target for drug discovery research. Based on existing findings, it can be concluded that beyond the currently employed chemotherapeutics, natural products (like flavonoids) exhibit pleiotropic, multi-target activities and are budding as possible complementary chemopreventive molecules against breast cancer with fewer side effects than conventional therapy. In this review, we comprehensively highlight an outline of the multiple pleiotropic pharmacological effects of various major classes of flavonoids on breast cancer with their specific mechanisms underlying its anticancer effect.

Use of different stains for microscopic evaluation for the diagnosis of Pythium keratitis.

PURPOSE: To evaluate the efficacy of various staining techniques for detection of Pythium in keratitis cases. METHODS: Data of nineteen consecutive culture-positive cases of Pythium keratitis were retrospectively analysed. Corneal scrapings and corneal buttons (in the cases which underwent therapeutic penetrating keratoplasty [TPK]) were sent for microbiological and histopathological examination. The direct smears were stained with Potassium hydroxide and calcofluor white (KOH â€‹+ â€‹CFW), Gram and Iodine-Potassium Iodide-Sulphuric Acid (IKI-H2SO4) stains. The corneal buttons were stained with Gomori's Methanamine Silver (GMS), Periodic Acid-Schiff (PAS) and Iodine-Potassium Iodide-Sulphuric Acid (IKI-H2SO4) stains. The positivity of various stains in detecting Pythium was studied. RESULTS: Gram and KOH â€‹+ â€‹CFW staining from smear was done in 16 out of 19 (84.2%) cases. KOH â€‹+ â€‹CFW and Gram stains were suggestive of Pythium in 10 (62.5%) and 7 (43.8%) cases, respectively. IKI-H2SO4 staining in scraping samples was positive for Pythium in all the 4 (100%) cases in which it was performed. Half corneal buttons were positive for Pythium with IKI-H2SO4 stain as well as GMS stain in all the 18 cases that underwent TPK (100%). PAS stain showed weak to faint pink staining of Pythium filaments in 7 out of 18 cases (38.9%). CONCLUSION: IKI-H2SO4 stain followed by KOH â€‹+ â€‹CFW stain detects Pythium filaments most accurately in corneal scraping samples from keratitis patients, although the differences were not statistically significant. The positivity of the stains depends on astute observation by an experienced ocular microbiologist and pathologist.

Identification of Candidate Immune System MicroRNAs Differentially Found in Colostrum and Milk Exosomes.

BACKGROUND: The fetus grows in a sterile womb environment. After birth, the newborn immune system has two immediate hurdles to clear. First immediate suppression of the womb compatible immune system and turn on the immune system of the newborn that can counter the antigenic world. The underlying mechanism of immune fluctuation by milk microRNAs (miRNAs) can be crucial for the treatment of critical or premature newborn. METHODS: We collected fourteen samples of each colostrum and mature milk from lactating mothers, four samples of each were used for microarray analysis, and the other ten were used for miRNA expression profiling by real-time PCR. RESULTS: From the microarray, 154 differentially expressed miRNAs were identified, whereas 49 miRNAs were revealed as immune-related miRNAs based on a literature study. Among the 49 miRNAs, 33 were already shown as strongly validated immune-related miRNAs (validated by qPCR, Western Blot, and Luciferase assay) and were considered for further analysis. Twenty-two miRNA expressions were analysed by real-time PCR as their Ct values were within considerable limits. Twelve numbers of miRNAs were significantly downregulated in mature milk compared to colostrum, which were again subjected to bioinformatics analysis to predict the biological mechanisms behind the differentially expressed miRNAs. CONCLUSION: This study shed light on the human milk exosome miRNA expression dynamics during lactation and their possible role in the gradual skewing of the newborns' immune system. The information is crucial for the development and onset of sepsis in premature newborns in the NICU.

Comparison of Corneal Tissue Profile of Pseudophakic and Phakic Donors.

PURPOSE: To compare the endothelial quality of corneas obtained from pseudophakic donors with age-matched phakic controls. METHODS: Retrospective analysis of 100 corneas each from pseudophakic and phakic eyes with donor age ≥60 years in both the groups was performed. The endothelial cell density, coefficient of variation, and percentage of hexagonal cells obtained by specular microscopy were compared between the two groups. The cut-off level of endothelial cell density (ECD) taken for optical keratoplasty was 2,000 cells/mm2. RESULTS: The male and female donors constituted 60% (n=120) and 40% (n=80), respectively. The mean age of the donors was 66.9±7.3 years in the phakic group and 69.9±7.7 years in pseudophakic group. The mean ECD in the phakic group was 2757.6±328.5 cells/mm2 and that in the pseudophakic group was 2225.5±471.9 cells/mm2 (P<0.0001). The mean coefficient of variation in the phakic group was 37.1±5.0 and that in the pseudophakic group was 38.6±11.1 (P=0.234). The mean percentage of hexagonality in the phakic group and the pseudophakic group was 52.0±6.4% and 51.2±7.2%, respectively (P=0.414). Both in univariate and multivariate linear regression analysis, age of the donor was found to be negatively associated in predicting ECD (P=0.002 and P=0.003, respectively). Sixty-nine and thirty-three corneas from the phakic and pseudophakic donor pool were used respectively. CONCLUSION: Difference in ECD between the phakic and the pseudophakic donor group was found to be statistically significant. The ECD in the pseudophakic group was found to be above the cut-off limit required for keratoplasty. Hence, pseudophakic corneas may also be used for transplantation.

Outcome of four-point fixated open loop polymethyl methacrylate anterior chamber intraocular lens.

PURPOSE: To study long-term visual and refractive outcomes and complications in eyes with anterior chamber intraocular lens (ACIOL) implantation. METHODS: Data of patients who underwent primary and secondary ACIOL implantation at L V Prasad Eye Institute, Bhubaneswar between 2011 and 2020 was collected, including details of post-operative visits. For analysis, sample was divided into: group Ia (primary ACIOL in cases without risk factors, n = 104); group Ib (primary ACIOL in cases with pre-existing risk factors, n = 49); and group II (secondary ACIOL, n = 40). RESULTS: A total of 193 eyes of 192 patients were included. Mean post-operative follow-up in groups I and II were 8.6 and 11.51 months, respectively. Mean pre-operative and last visit corrected distance visual acuity were 1.73 ± 0.11 and 0.42 ± 0.05 logMAR units in group Ia (p < 0.001), and 1.53 ± 0.14 and 0.49 ± 0.10 logMAR units in group Ib (p < 0.001). The mean spherical equivalent (MSE) for last refraction was -0.37 ± 0.18 diopters (D) and -0.15 ± 0.51 D in groups I and II, respectively. Of 76 eyes in which addition of 2.5 D (over the near emmetropic posterior chamber intraocular lens power) was taken for ACIOL, 40 (52.6%) had MSE within ± 0.5 D. Most common complications were transient corneal edema and anterior chamber reaction. Eyes on anti-glaucoma medications at last visit were eight (7.7%), 15 (30.6%), and two (5.0%) in groups Ia, Ib, and II, respectively. CONCLUSION: We observed that ACIOLs have good visual and refractive outcomes. Raised IOP is a concern in eyes with pseudoexfoliation, but can be managed with close monitoring. Hence ACIOL can be a good option for managing aphakia after cataract surgery.

Determining Theileria annulata parasitemia through qPCR in lactating cows of Odisha, - a theileriosis-endemic region of India.

Study was undertaken in a theileriosis-endemic region of India during May 2018 to April 2019 among milch cows. Blood samples collected from apparently healthy (n = 65) and Theileria-suspect cows (n = 65) were screened against T. annulata and T. orientalis infection by SYBR Green‒based real time PCR using primers designed from the isolates of study area. Cows having single infection with T. annulata with/without clinical signs of inappetence, low milk yield, pale mucous membranes, fever, enlarged prescapular lymph node, soil licking, panting, coughing, salivation and lachrymation were subjected to further investigation where parasitaemia and piroplasms per 1000 erythrocytes ranged from 1.6 × 107 to 1.2 × 108 parasites/mL of blood and 3-24 piroplasms in moderate group (16/65), 4.4 × 108 to 6.9 × 109 parasites/mL of blood and >88 piroplasms in severe group (30/65) and 1.6 × 104 to 5.5 × 106 parasites/mL of blood and 0-1 piroplasms in asymptomatic or carriers (17/65), respectively. Study unfolded significant difference in T. annulata parasitaemia among apparently healthy and ill cows. Phylogenetic analysis of our T. annulata isolates (NCBI accession numbers MN098316, MN098317 and MN098318) exhibited maximum similarity with the isolates detected in other parts of India.