Comparative molecular characterization and phylogenetic analysis of cerebral and non-cerebral coenurosis in Indian goats.

Coenurus cerebralis is the larval stage of Taenia multiceps, which infects the muscles and brain of goats and, to a lesser extent, sheep. The resulting cerebral and non-cerebral infections caused by the larval form (metacestode) of this cestode are commonly known as coenurosis. A weak emaciated carcass of five months old female goat, on necropsy, revealed numerous parasitic cysts (n = 56, grossly visible) in the visceral cavity including heart, diaphragm, thoracic cavity, abdominal cavity and pelvic inlet. A large number of variable sized parasitic cysts were also observed embedded in the pericardium and myocardium causing functional damage to the heart. The parasite caused extensive tissue damage at gross and microscopic levels in the heart including traumatic destruction of the myocardium with degenerative and necrotic changes and infiltration of mononuclear cells. On parasitological examination, the cysts were identified as Coenurus cerebralis, as the scolices had characteristic four suckers and a rostellum with a double crown of hooks. Further confirmation was done using polymerase chain reaction targeting specific ND1 and CO1 genes. Phylogenetic analysis of CO1 and ND1 genes showed a major branch comprising two clades of T. multiceps grouped as separate entities with the first clade showing T. multiceps/Coenurus cerebralis native CIRG strain (cerebral) being placed in proximity to T. multiceps/Coenurus cerebralis CIRG strain (non-cerebral/visceral) compared to the Chinese strains of T. multiceps. The phylogenetic analysis of ND1 and CO1 genes of C. cerebralis of cerebral and non-cerebral isolates revealed close proximity but expressed in two different disease forms (i.e., visceral coenurosis and neural coenurosis) which indicated that they were very close divergent from a common ancestor. On the basis of the observations it was concluded that goat died due to cardiac dysfunction resulting from severe systemic infection of metacestode of T. multiceps was closely related to isolate that caused neural coenurosis in another goat. Based on the sequencing analysis and phylogenetic information, the possible differences in the clinical manifestation (neural or visceral) could be attributed to the pathogenesis.

Eucalyptus robusta leaves methanolic extract suppresses inflammatory mediators by specifically targeting TLR4/TLR9, MPO, COX2, iNOS and inflammatory cytokines in experimentally-induced endometritis in rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Bacterial endometritis is one of the major causes of reproductive disorders including infertility in farm animals. Antibiotics are generally used for treatment of such disorders but now a days residues of antibiotics are of great public health concern, therefore, phytoremediation is being considered as an alternative to use of antibiotics. AIM OF THE STUDY: Present study was undertaken to investigate the efficacy of Eucalyptus robusta leaves methanolic extract against endometritis along with the possible mechanism of action especially targeting inflammatory biomarkers. MATERIALS AND METHODS: Bacterial endometritis was produced using clinical isolates of E. coli and Staphyloccocus aureus from bovines (cows and buffaloes) endometritis cases. After seven days of inoculation of the mixed bacterial culture, endometritis was confirmed based on the presence of visible pus and edema, thinning of endometrial lining and presence of large number of polymorphonuclear cells and bacterial load in uterine flushing. Female Wistar rats were divided in to five groups namely control, sham-operated, endometritis, endometritis plus Eucalyptus leaves extract and endometritis plus cefixime. Serum specific inflammatory biomarkers (interleukin-1ß, interleukin-10, tumor necrosis factor-α, intercellular adhesion molecule-1, serum amyloid A) and myleoperoxidase, toll like receptors-4 and -9, inducible nitric oxide synthase, nitric oxide, cyclooxygenase 1 and 2 were estimated in uterine tissues using ELISA kits. RESULTS: Interleukin-10, serum amyloid A, myleoperoxidase, toll like receptors-4 and-9, cyclooxygenase-2, inducible nitric oxide synthase and nitric oxide were significantly increased while non significant increase in interleukin-1ß, cycloxygenase-1 and intercellular adhesion molecule-1 were observed but level of tumor necrosis factor-α was found decreased in rats of endometritis group. Histopathological lesions in uterus showed efficient induction of endometritis by presence of inflammatory cells which are lessened effectively after treatment with Eucalyptus leaves extract. Eucalyptus robusta leaves extract produced curative and protective effect against endometritis and results were comparable to or even better than cefixime. CONCLUSIONS: Eucalyptus robusta leaves extract possess promising antibacterial activity and efficacy against experimental endometritis and, therefore, holds promising potential for development of effective formulation for treatment of endometritis in animals.

Fast-dissolving ocular films of riboflavin acetate conjugate for treatment of keratoconus in UVA-CXL procedure: ex vivo permeation, hemolytic toxicity and apoptosis detection.

BACKGROUND: Attempts to facilitate corneal epithelial penetration of riboflavin (Rb) without de-epithelization, so far, include the use of penetration enhancers, to devitalize corneal epithelium in order to disturb tight epithelial interjunctional complexes and zonulae occludentes. Though such approaches result in sufficient epithelial permeability of Rb to guarantee efficacy of CXL procedure, they lack the evidences of safety. Prodrug with improved lipophilicity targeted toward esterases and amidases has proven to be an effective and promising approach to overcome lipophilic corneal epithelial barrier. OBJECTIVES: Fast-dissolving ocular films of newly synthesized and characterized riboflavin lipid conjugate (RbLDC) were developed to overcome corneal epithelial barrier resistance for treatment of keratoconus. The safety concern of the film was assessed by in vitro hemolytic toxicity and in vitro apoptosis detection for its safe clinical use. RESULTS: The optimized film was tough, flexible and dissolved rapidly within 36.86 s in simulated tear fluid, pH 7.4. FE-SEM/EDX showed smooth surfaces of films and evidenced the quantitative elemental similarity, indicating drug homogeneity. The permeation profile of F18 demonstrated 13.28-fold increased permeation of RbLDC relative to Rb solution across intact cornea. Safety was confirmed by 3.74% hemolysis and 10% apoptosis. CONCLUSION: Safe and efficient RbLDC fast-dissolving ocular films capable of overcoming corneal epithelial barrier resistance to avoid surgical intervention of corneal epithelial debridement were developed.