First report of molecular and phylogenetic analysis of Physaloptera praeputialis in naturally infected stray cats from India.

Nematodes of the genus Physaloptera are globally distributed and infect a multitude of hosts. Their life cycle involves orthopterans and coleopterans as intermediate hosts. The morphological characters alone are inadequate to detect and differentiate Physaloptera spp. from its congeners. Moreover, molecular studies are limited to compare them precisely. The present communication reports the first molecular phylogenetic characterization of feline Physaloptera spp. from India based on mitochondrial cytochrome c oxidase subunit 1 (COX1) and small subunit ribosomal DNA (18S rDNA). The nematodes were first isolated from the stomach of adult stray cats during necropsy examination. Based on the gross and microscopic characters, the worms were identified as P. praeputialis. Morphological identification was further confirmed through PCR targeting the barcode region of the mitochondrial cytochrome c oxidase subunit I (MT-COI) gene, using nematode-specific primers cocktail followed by species specific primers targeting partial COX1 and 18S rRNA genes. Generated sequences were submitted in NCBI GenBank (MW517846, MW410927, MW411349), and phylogenetic trees were constructed using the maximum likelihood method. When compared with other sequences of Physaloptera species across the globe, the present isolates showed 85.6-97.7% and 97.3-99% nucleotide homology based on COX1 and 18S rRNA gene, respectively. BLASTn analysis revealed a strong identity to other Physaloptera spp., and the phylogenetic tree placed all Physaloptera spp. in the same cluster. This study again indicates the usefulness of molecular techniques to substantiate the identity of species that may lack adequate descriptions and impart new insight for the potentially overlooked significance of P. praeputialis infections in felines.

First report of the isolation and phylogenetic characterization of equine Setaria digitata from India based on mitochondrial COI, 12S rDNA, and nuclear ITS2 sequence data.

Equine ocular setariasis arising mainly from ectopic infestation of Setaria digitata is a common vision impairing ophthalmic disease in India, and the identification of this filarial nematode is based solely on morphology. However, morphological characters alone are inadequate to detect and differentiate S. digitata from its congeners. The present communication reports the first phylogenetic characterization of equine S. digitata from India based on sequences derived from the mitochondrial cytochrome c oxidase subunit 1 (COI), the mitochondrial small subunit ribosomal DNA (12S rDNA), and the nuclear internal transcribed spacer 2 (ITS2). Three isolates were characterized for each gene, and respective sequences were submitted to NCBI database (MN078131, MN078132, and MN095798). The sequences were also compared with the other related sequences available from PubMed around the globe, and phylogenetic analysis was carried out in conjunction with nucleotide homologies. There was no intraspecific variation among the Indian isolates. The phylogenetic analysis of S. digitata, inferred from these genes, showed that the isolate sequences obtained from different host species created a separate monophyletic clade within the genus Setaria with minor sequence variations revealing similar molecular characteristics of S. digitata isolates throughout the globe. In addition, the studied Indian isolates were found closer to Sri Lankan isolates. The S. digitata and S. labiatopapillosa appeared as sister species.

Hematobiochemical alterations and direct blood polymerase chain reaction detection of Theileria annulata in naturally infected crossbred cows.

AIM: The aim was to determine hemato-biochemical changes and rapid diagnosis of Theileria annulata in naturally infected crossbred cows. MATERIALS AND METHODS: Blood samples from lactating crossbred cows (n=40) between 3 and 7 years of age and showing clinical signs of tropical theileriosis were collected, with or without anticoagulant, and analyzed for tropical theileriosis by direct smear, direct blood polymerase chain reaction (PCR) detection of merozoite-piroplasm surface antigen (Tams1) gene specific amplicon, estimation of hematological and biochemical parameters. Healthy crossbred cows (n=6), examined free from hemoprotozoan infections were included as control. RESULTS: The infected crossbred cows revealed significantly (p<0.001) lower values of total erythrocytic counts (4.46±0.2 × 10(6)/µL), hemoglobin (Hb 6.025±0.39 g%), packed cell volume (17.05±1.1%), mean corpuscular volume (37.94±1.70 fL) and mean corpuscular Hb (13.5±0.48 pg; p<0.002) compared with healthy control. The serum samples of infected cows revealed profound (p<0.05) hyponatremia (Na 133.21±2.36 mEq/l) and hypocalcemia (Ca 8.39±0.34 mg%). Infected crossbred cows showed a significant increase (p<0.05) of mean serum activity of alanine aminotransferase (61.45±13.36 U/L), aspartate aminotransferase (146.1±20.97 U/L), blood urea nitrogen (28.26±3.90 mg%), creatinine (1.55±0.13 mg%), direct bilirubin (0.33±0.04 mg%; p<0.001) and lactate dehydrogenase (3001.32±167.0 U/L; p<001). Blood direct PCR revealed a 721-bp fragment amplified from the target gene encoding 30-kDa major merozoite surface antigen of T. annulata using specific primer pairs. This assay was positive for all the infected animals. CONCLUSION: The assessments of hemato-biochemical parameters in T. annulata infected crossbred cows may be useful in understanding disease pathogenesis, prognosis and corrective measures for supportive therapy. Moreover, blood direct PCR can reliably be used for rapid detection of T. annulata in conjunction with microscopic examination.

Auto-immunological basis of disk degeneration.

Ninety-four subjects were divided into 5 groups according to symptoms and radiologic findings to devise a test for immunoglobin levels and lymphocyte microstimulation. Cellular immunity to disk antigens can be demonstrated in disk degeneration which may be primary or secondary. The serum level of IgM, irrespective of the age of the individual was significantly higher in patients with backache than asymptomatic patients.

Coccidioidomycosis of bone and joints.

Sixteen cases of coccidioidomycosis involving bone and joints were treated with Amphotericin B. In all systemic or disseminated cases Amphotericin B should be prescribed. Local Amphotericin B is useful in joints if the disease is limited, and in patients who do not tolerate Amphotericin B by systemic route. If the patient cannot be treated with systemic Amphotericin B, surgical treatment should be directed to remove all possible infected tissue. Patients with a negative skin test and albuminuria in disseminated coccidioidomycosis should be treated with Amphotericin B and passive immune transfer.

Transfer of tibialis posterior tendon in cerebral palsy.

Posterior tendon transfer to the dorsum of the foot through the interosseous membrans is an effective procedure for spastic equinovarus deformity. The complications of excessive calcaneal or excessive valgus deformity can be avoided if the transfer is not inserted too tightly with the foot in dorsiflexion and if it is not transferred to a cuboid insertion. Electromyographic activity was demonstrated in some of the transferred muscles during the swing phase of gait. Tendo achillis lengthening in association with posterior tibial transfer must be done judiciously to avoid deformity of the calcaneus.

Fat embolism following bilateral total knee replacement with the Guepar prosthesis. A case report.

Fat embolism followed bilateral total knee replacement with Guepar prostheses in a rheumatoid patient the day after the operation. The risk of fat embolism following total knee replacement with a hinge prosthesis is especially high in rheumatoid patients with compromised pulmonary functions and a history of long-term treatment with corticosteroids.

Total hip replacement in patients with ankylosing spondylitis with involvement of the hip.

Twenty-three patients with ankylosing spondylitis underwent thirty-four total hip arthroplasties with an average follow-up of 42.5 months. The over-all results were rated good in all but two patients. Significant benefit was obtained in posture and function so that these patients could be gainfully employed. Improvement in the sum total range of motion was 96.1 degrees in patients with bone ankylosis and 150 degrees in those with fibrous ankylosis. The preoperative sum total range of motion was maintained or improved in hips with a painful arc of motion. Most of the limited motion in patients with ankylosed hips was due to myositis ossificans.

Acute vascular compression of the duodenum following cast application.

It is suggested that only those patients in whom symptoms begin within two weeks after application of the cast should be included in the cast syndrome. Early diagnosis with prompt treatment in the form of replacement therapy and a nasogastric tube is important. Operation should be reserved for those patients in whom conservative treatment fails or the cast cannot be removed.