Significantly increased recovery of intestinal parasites on routine stool specimen evaluation.

Three hundred thirty-six stool samples from October 2001 through October 2002 were analyzed for the presence of intestinal parasites. Fifty-six of these (16.7%) were positive for a total of 66 parasites; 65/66 (98.5%) were detected by iodine and dimethyl sulfoxide-modified acid-fast (DMSO-mAFB) stained smears of fresh and formalin-ethylacetate sedimentation concentrated samples. Saline, iodine, and DMSO-mAFB stained smears of fresh stool samples alone detected significantly fewer parasites, finding only 50/66 (75.8%) (p < 0.05). Stool samples analyzed by trichrome stained specimens preserved in Zinc sulfate polyvinyl alcohol (Zinc PVA) detected only 41/ 66 (62.2%) of the parasites. In our study population, it was necessary to perform the National Committee for Clinical Laboratory Standard (NCCLS) recommended to accurately detect intestinal parasites. The concentration technique is simple and significantly increased the detection of intestinal parasites.

Gene encoding the group B streptococcal protein R4, its presence in clinical reference laboratory isolates & R4 protein pepsin sensitivity.

BACKGROUND & OBJECTIVES: R proteins were first identified by Lancefield in group B Streptococcus (GBS) as resistant to trypsin at pH8 and sensitive to pepsin at pH2. The R4 protein found predominantly in type III and some type II and V invasive isolates conforms to these criteria. The Rib protein, although structurally and epidemiologically similar to R4, was reported as resistant to both proteases. We report here the gene encoding the R4 protein from a type III group B streptococcal isolate (76-043) well characterized in our laboratory. METHODS: Trypsin extracted GBS proteins were assayed for protease sensitivities by double-diffusion Ouchterlony using varying conditions for the enzyme pepsin. Standard haemoglobin assay was used to examine pepsin enzymatic activity. Thirty clinical isolates of varying protein profiles identified by double-diffusion from our reference strain laboratory were screened by PCR and Southern technique. SDS-PAGE gel purified R4 amino acid sequences were determined and used to design oligonucleotide primers for screening a 76-043 genomic library. RESULTS: R4 was sensitive to pepsin at pH2 but appeared resistant at pH4, the reported pH used for Rib. By standard haemoglobin assay and trypsin extract studies of R4 protein, pepsin was shown to be active at pH2, yet easily inactivated; assays of GBS surface proteins are critical at pH2. Of the amino acids initially sequenced from R4, 88 per cent (61/69) showed identity to Rib; the r4 nucleotide sequence was identical to that of rib. All isolates with strong positive protein reactions for R4 were positive in both PCR and Southern technique, whereas isolates expressing alpha, beta, R1/R4, and R5 (BPS) protein profiles were not. INTERPRETATION & CONCLUSION: Sequenced PCR products aligned with identity to the R4 and Rib nucleotide sequences and confirmed the identity of these proteins and their molecular sequences.

Effect of bracken (Pteridium a quilinum) and dryopteris (Dryopteris juxtaposita) fern toxicity in laboratory rabbits.

Experimental studies with Bracken and Dryopteris ferns @ 25% concentrate ration mixture were conducted in rabbits. Fern fed rabbits showed progressive anaemia, leukopaenia, lymphopaenia and relative heterophilia. Significant elevations in serum enzymes like serum glutamate oxaloactate transminase (SGOT), serum glutamate pyruvate transminase (SGPT), alkaline phosphatase (ALP), urea and creatinine levels were seen. Histopathologically, rabbits showed mild to moderate vascular changes in most of visceral organs, vacuolar degenerative changes in hepatocytes, hypersecretory activity in intestine, presence of casts in renal tubules and degenerative changes in renal tubular lining epithelial cells. Dryopteris fed rabbits showed somewhat more severe degenerative and vascular changes in different intervals. A low level of toxic principle ptaquiloside was detected in Bracken and Dryopteris ferns by thin layer chromatography (TLC) and high-pressure liquid chromatography (HPLC) methods.