Short Variable Regions flaA Gene (SVR-flaA) Diversity and Virulence Profile of Multidrug-Resistant Campylobacter from Poultry and Poultry Meat in India.

Human gastrointestinal infections caused by Campylobacter species is the second most important foodborne illness after salmonellosis worldwide. Poultry represent one of the main sources of Campylobacter organisms. In the present study, the short variable region of flagellin gene (SVR-flaA) typing was carried out to determine the variation among the circulating strains of Campylobacter jejuni and Campylobacter coli. The C. jejuni and C. coli isolated from poultry and poultry meat were screened for the presence of virulence determinants like cadF, flaA, cdtB, and wlaN gene. The screening for wlaN gene is crucial in view of the fact that most patients with Guillian Barre's (GB) syndrome with a preceding history of diarrheal illness have been found to harbor wlaN gene-positive C jejuni strains. Out of the 200 samples comprising poultry meat and cloacal swabs, 21.5% of samples were found to harbor Campylobacter spp. of which 2.5% were Campylobacter jejuni, and 19% were confirmed as Campylobacter coli. The cadF, flaA, cdtB virulence genes were detected in all the Campylobacter spp. isolated in the present study. The presence of the wlaN gene in the Campylobacter jejuni isolated in the present study may pose a public health threat with long-term human health implications. The SVR-flaA typing of twelve Campylobacter isolates obtained in the present study revealed that Campylobacter coli flaA sequence OL471375 is a new strain with a novel allele type 1,675 and peptide sequence 5 which stands deposited in pubMLST database for Campylobacter. The other flaA-SVR gene sequences identified in this study were OL471369, OL471370, OL471371, OL471372, OL471373, and OL471374. Among twelve Campylobacter spp., three distinct DdeI-RFLP patterns were observed, each varying in size from 100 to 1,000 base pairs. Antimicrobial profiling of the Campylobacter spp. isolated in the present study revealed that 50% of the strains were multidrug resistant. All the Campylobacter spp. were resistant to ciprofloxacin (CIP), ampicillin (AMP), penicillin (PEN), and nalidixic acid (NAL) whereas 57.1% of strains were resistant to tetracycline (TET) and erythromycin (ERY) 28% to amoxicillin (AMX) and enrofloxacin (ENO), 85% to amikacin (AMK). The high degree of resistance to fluoroquinolones observed in the present study is crucial in view of fluoroquinolones being drugs of choice for the treatment of human Campylobacter infections.

Orbital Mucormycosis: Understanding the Deadly Fungus Sweeping the Globe.

Introduction Mucormycosis (black fungus) is a rare opportunistic fungal infection commonly affecting immunocompromised individuals. There has been a surge in the number of these cases during the second wave of the coronavirus disease 2019 (COVID-19) in India. Mucormycosis has been reported to occur within a week or a few weeks post-recovery from COVID-19. The most common clinical manifestation of mucormycosis is rhino-orbital-cerebral mucormycosis (ROCM). At our tertiary care center, we initiated a prospective study to identify risk factors, study ocular manifestations, and explore medical and surgical management of orbital mucormycosis patients in the post-COVID-19 era. Material and methods This is a detailed description of a prospective observational hospital-based study. The study included 148 patients who presented with ROCM. A detailed history was taken regarding the complaint, duration, and associated risk factors. Systemic, local, and complete ophthalmic examinations were done that included assessment of extraocular movements, visual acuity, slit-lamp examination, and fundus examination. All data were recorded separately for each patient in a pre-decided proforma. Result The study group consisted of 148 patients. In our study, the highest association was with COVID-19-positive status (68.24%), out of which 57 (56.43%) were on oxygen support. Diabetes mellitus contributed next to COVID-19 with 86 (58.10%) patients with a positive history of diabetes. Seventy-one (47.97%) patients were on steroids, out of which 68 (67.32%) were COVID-19-positive and the rest (23%) were on steroids due to various systemic reasons. Rhinomaxillary involvement was present (51%). Out of 63 patients with orbital involvement, 16 (25.39%) presented bilaterally and 47 showed unilateral orbital involvement more on the right side (42.85%). The predominant location of orbital involvement was the orbital apex. The most common symptom seen in our study was nasal discharge (86.5%), and ophthalmoplegia was the most common sign. Conclusion Corticosteroids should be used with caution to prevent negative impact and potential ROCM. Good glycemic and metabolic control is crucial for treatment. Management of mucormycosis involves surgical debridement, antifungal agents, and retrobulbar amphotericin B injections. Early diagnosis and aggressive treatment are essential for success. Orbital exenteration may be necessary for advanced stages, while conservative approaches may work for earlier stages. Patient counseling is needed for cosmetic rehabilitation. A multidisciplinary approach involving various specialists is necessary.

Phacoemulsification in phacomorphic angle closure: Our experience.

PURPOSE: To evaluate intraocular pressure (IOP) control, visual prognosis, and complications following phacoemulsification in eyes with phacomorphic angle closure of <10 days' duration. METHODS: Prospective, nonrandomized interventional consecutive case series included all patients with phacomorphic glaucoma who presented between November 2020 and November 2021. All patients underwent slit-lamp biomicroscopy, applanation tonometry, and gonioscopy of the other eye. Phacoemulsification with IOL implantation under topical anesthesia was performed in all cases. A complete ophthalmic examination was done at each follow-up visit. RESULTS: A total of 50 eyes with phacomorphic glaucoma were included in this study. The preoperative mean IOP was 41.12 ± 8.20 mmHg and the mean IOP at last visit was 13.84 ± 2.08 mmHg. There was a statistically significant difference between IOP at presentation and IOP at last follow-up (P < 0.001) (Wilcoxon Signed-Ranks Test). There was no requirement of long-term anti-glaucoma medications in any patients. No significant intraoperative complications were noted. The final postoperative best-corrected visual acuity was 6/12 or better in 32 patients. Sixteen eyes had corneal edema and 20 eyes had anterior chamber inflammation on postoperative day one that resolved with standard medical therapy. CONCLUSION: Phacoemulsification is safe and effective in controlling IOP and achieving good functional visual acuity with minimal complications in the management of phacomorphic glaucoma in expert hands. Our study also lays emphasis on public awareness, early detection, and management of advanced cataract, so that the incidence of this potentially blinding entity can be reduced.

Clostridium perfringens from fresh water fish of Kashmir Himalaya and their aquatic environment: Toxinotyping and phylogenetic analysis.

Clostridium perfringens is a common anaerobic foodborne pathogen known to produce >20 toxins. In nature, this bacterium has 7 different toxinotypes (A-G) based on the presence of its 6 main toxins. The present study examined the occurrence of different toxinotypes of this bacterium in the ichthyofauna and aquatic environments of Kashmir Himalayan lakes, India. A total of 510 samples (210 water; 150 each of common carp and snow trout) were collected from 3 different lacustrine habitats (Dal, Anchar and Nigeen Lakes) of the region. By performing 16S rRNA PCR test, it was observed that all 210 water samples and 80 (26.66%) of 300 fish samples tested for this specific bacterial species were positive. Then by using multiplex-PCR targeting six virulence genes of C. perfringens, it was confirmed that all the 290 isolates from water and fish samples were positive for Toxinotype A, as only cpa toxin gene was amplified. Phylogenetic analysis of the amplified gene and its amino-acid sequences revealed 95%-98% homology with analogous sequences of this bacterial strain reported from China, Egypt and India. The study documents the existence of C. perfringens toxinotype A in the ichthyofauna of Kashmiri Himalayan lakes, entailing that fish can likely act as transmission medium for C. perfringens food poisoning to humans via food.

Human immune deficiency virus conundrum: An everlasting challenge!

BACKGROUND: Human immune deficiency virus (HIV) causes a wide spectrum of diseases worldwide and has the capability to affect every organ system in the body. Ocular manifestations have been reported in up to 70% of individuals infected with HIV and the ocular manifestations reflect systemic disease and maybe the first sign of disseminated infection. PURPOSE: To study the pattern of ocular manifestations related to HIV in seropositive patients with the objective of identifying its spectrum in highly active antiretroviral therapy (HAART) era and to emphasize on the role of ophthalmologists in fighting the battle against HIV. MATERIALS AND METHODS: A cross sectional study was undertaken on 300 HIV positive patients. Data were collected, tabulated and analyzed using MSTAT software. Chi-square test was applied and P < 0.05 was considered significant. RESULTS: Ocular lesions were observed in 50.33% of patients. Conjunctival microvasculopathy being the most common finding seen in 27 (9%) patients, followed by retinal microvasculopathy and trichomegaly in 20 (6.66%) patients each. Herpes zoster ophthalmicus and cytomegalovirus retinitis were observed in four (1.33%) and three (1%) patients respectively. In the study 78.33% patients were on ART. CONCLUSION: HAART has reduced serious ocular opportunistic infections and ocular malignancies, but HAART mediated visually disabling immune recovery uveitis has emerged as a clinical challenge for ophthalmologists. Improved communication between the two broad specialties of HIV medicine and ophthalmology will definitely go a long way in the battle against this dreadful disease.

Genome sequence of Dichelobacter nodosus JKS-07B isolate from J&K, India associated with virulent footrot of sheep.

INTRODUCTION: Virulent footrot of sheep caused by Dichelobacter nodosus is associated with tremendous economic losses due to recurrent treatment costs and increased culling rates. This organism being a fastidious anaerobe is difficult to isolate on ordinary media that does not support its growth. The D. nodosus serogroup B isolate described in the present study has been used in the preparation of the whole-cell killed vaccine against footrot in India. D. nodosus serogroup B is the predominant serogroup involved in virulent footrot (lesion score 4) in India as well as in many sheep-rearing countries of the globe. METHODS: Genomic DNA was extracted using wizard Genomic DNA purification kit. The whole genome of the D. nodosus strain B was sequenced using an Illumina HiSeq 2500 platform and annotated according to functional gene categories. Annotations were performed using in-house developed Perl scripts using Nr/Nt database, uniprot, Pfam, KEGG, Panther DB, and GO database. RESULT: The assembled genome size is 1.311,533 Mb and GC content is 44.38. A total of 1215 protein-coding genes, 44tRNA and 7 rRNA were identified. The genome shows 98.63% sequence homology with the reference genome. However, 21 new genes have been identified in this genome. The information will provide insights into the various genes and regulators necessary for D. nodosus growth and survival. DISCUSSION: The genome information of this serogroup B of D. nodosus isolate involved in 85-90% cases of virulent footrot of sheep in India provides further insights for improvement of the killed vaccine (B serogroup) developed recently in India. For the development of an efficacious vaccine against virulent footrot, it is essential to know the serological diversity as well as the virulent status of the strains of the D. nodosus. This serogroup isolate is a potential vaccine candidate to mitigate ovine footrot in India as the majority of virulent footrot cases belong to serogroup B of D. nodosus.

Methicillin resistance genes and in vitro biofilm formation among Staphylococcus aureus isolates from bovine mastitis in India.

INTRODUCTION: Biofilms, an assemblage of microbial cells irreversibly associated with a surface and enclosed in a matrix of polysaccharide material pose serious health challenges, resulting in high economic losses. The emergence of methicillin-resistant S. aureus (MRSA) infections and ability to form biofilms in dairy animals is of emerging concern for livestock and public health owing to their association with serious infections. The present study was undertaken to examine the presence of methicillin resistance genes among the biofilm forming Staphylococcus aureus strains isolated from cases of acute and subacute bovine mastitis. A total of 150 mastitic milk samples referred to Veterinary Clinical Complex, Shuhama (Aulesteng) SKUAST-K were screened in present study. The methicillin resistant Staphylococcus aureus isolates were also screened for in vitro biofilm forming ability. RESULTS: A total of 80 (53.33%) S. aureus isolates were recovered from cases of bovine mastitis of which 20 (25%) were methicillin (mecA) gene positive. Of the 20 mecA positive isolates, 20% were positive for SCCmec I, 35% for SCCmec IV and 45% for SCCmec V subtypes. In vitro antibiotic sensitivity testing of MRSA revealed complete resistance towards methicillin and other pencillin group of antibiotics. CONCLUSION: A significant correlation was observed between in vitro biofilm formation and presence of methicillin resistance gene in S aureus isolates recovered from acute and subacute mastitis. The Staphylococcus aureus isolates positive for methicillin resistance gene (mecA) were either strong or moderate biofilm formers.

Draft genome sequence of Dichelobacter nodosus JKS-07 serogroup E from India.

OBJECTIVES: Dichelobacter nodosus is an anaerobic bacterium with fastidious growth requirements that is the principal cause of footrot associated with lameness in sheep and goats. In India, D. nodosus serogroups B and E have been recorded as major causes of footrot. Here we report the draft genome sequence of a D. nodosus serogroup E strain (JKS-07) from a case of virulent footrot in India. METHODS: The whole genome of the D. nodosus JKS-07 serogroup E was sequenced using an Illumina HiSeq 2500 platform and was annotated according to functional gene categories. De novo genome assembly and annotation were performed using Perl scripts developed in-house using the Nr/Nt and UniProt databases. RESULTS: The assembled genome is 1389350bp and contains 1301 genes. The genome has 45 tRNAs and 9 rRNAs. The draft genome sequence will provide insight into the various genes and regulators involved in D. nodosus growth and survival. CONCLUSION: Information on the genome of the D. nodosus serogroup E strain is important bearing in mind the fact that both serogroups B and E are associated with virulent footrot, either alone or frequently together. In order to develop an efficacious vaccine against virulent footrot, it is essential to know the serological diversity as well as the virulence status of the D. nodosus strains. Serogroups B and E are potential vaccine candidates to mitigate ovine footrot in India.

The detection of Dichelobacter nodosus and Fusobacterium necrophorum from ovine footrot in Kashmir, India.

In a study conducted, a total of 450 swab samples from footrot lesions of naturally infected sheep were collected in all the ten districts of the Kashmir valley and were examined for the presence of Dichelobacter nodosus (D. nodosus) and Fusobacterium necrophorum (F. necrophorum), in order to determine if F. necrophorum was associated with ovine footrot. The detection of F. necrophorum and D. nodosus was carried out by polymerase chain reaction targeting the leukotoxin (lktA) and 16S rRNA genes, respectively. In this study, only less than 50% of positive samples contained both the bacteria, so it is not possible to conclude with certainty that both bacteria are together required for the disease manifestation.