Spermatozoal ultrastructure of four Sparidae fishes: Acanthopagrus berda, Acanthopagrus australis, Lagodon rhomboids and Archosargus probatocephus.

The mature spermatozoa of two Taiwan protandrous hermaphrodite Sparidae Acanthopagrus berda and Acanthopagrus australis are investigated and compared with those of other two Sparidae (Lagodon rhomboids and Archosargus probatocephus) from the Western hemisphere. Ultrastructurally the spermatozoon of these four species has a spherical, homogeneously electron-dense nucleus with an axial nuclear fossa. The midpiece contains one to four spherical mitochondria and encircles the basal body of the flagellum. The mature spermatozoa of the four species are of the primitive or ect-aquasperm form and conform to the teleostean type I spermatozoon with the flagellar axis inserts perpendicular and medial to the nuclear fossa. Variation in the depths of the nuclear fossa and mitochondria number is substantial in these four Sparidae species. This study provide useful systematic characters to the existing knowledge of comparative spermatology of Sparidae.

Ultrastructure of Pagrus major and Rhabdosargus sarba spermatozoa (Perciformes: Sparidae: Sparinae).

Transmission and scanning electron microscopy were used to investigate the ultrastructure of spermatozoa in two Sparinae species Pagrus major and Rhabdosargus sarba. Ultrastructurally, the spermatozoa of P. major and R. sarba both consist of a spherical, homogeneously electron-dense nucleus with a deep axial nuclear fossa, and an unusual notch, in the nuclear region. The midpiece contains two spherical mitochondria in R. sarba and one in P. major. The comparison of spermatozoal ultrastructure of these two species of Sparidae shows that they closely resemble one another and suggests that they are closely related. Variation in the geometry and dimensions of the mitochondrion and nucleus is substantial in these two Sparidae species. It is concluded that the spermatozoa of both species are of primitive type, and they are distinguished by several unique features which may provide useful systematic characteristics.

Identification of the human crooked neck gene by comparative gene identification.

Drosophila melanogaster serves as a useful model organism for functional genomic studies, and its genome project was recently completed. We previously described a comparative-gene-identification approach to assist human ortholog gene identification that involves applying an entire proteome as an alignment template. Analysis of the available 14100 Drosophila protein sequences revealed that 37% of them (5228 sequences) might lead to discoveries of novel human genes. Upon further database interrogations, we found several putative full-length human gene transcripts, including the human crooked neck (crn) gene. Based on sequence gap-closure experiments using reverse transcriptase-polymerase chain reaction as well as bioinformatic analysis, we found that the assembled human cDNA contig of crooked neck gene was at least 3903 base pairs in length with alternative splicing variations which encoded mainly for a 687-amino-acid residue protein. The human crooked neck gene was located on chromosome 20 with at least 15 exons. The unique features of the 16 copies of the tetratrico peptide repeat (TPR) motif were conserved in the yeast, fly and human crooked neck orthologous proteins, which were important for spliceosome assembly in cells.

Quantitative evaluation of pork adulteration in raw ground beef by radial immunodiffusion and enzyme-linked immunosorbent assay.

Quantitative estimates are important to establish whether pork adulteration in ground beef is accidental or intentional. A standard agar gel radial immunodiffusion (RID) test using forensic-grade antiserum to porcine albumin and an enzyme-linked immunosorbent assay (ELISA) using forensic-grade anti-porcine glycoprotein immunoglobulin were used to determine from 1 to 75% raw pork in raw ground beef. The RID test, which incorporated 1.5% anti-pork serum in 1% immunodiffusion agar, formed precipitin rings with pork albumin in agar wells. A linear standard curve was obtained by plotting the diffusion area against standard pork concentrations ranging from 0 to 80%. For the ELISA the endpoint optical density increased linearly versus log % pork between 0.0625% and 2% pork. In spiked samples, the RID test had a detection limit of 3 to 5%, a coefficient of variation (CV) of 22%, and a recovery of 105%. The ELISA had a detection limit of 1%, a CV of 18%, and a recovery of 114%. The mean recovery from the spiked samples by the ELISA and RID test was not significantly different (P > 0.05) from the known sample amounts. Quantitation by RID of 28 ground beef samples (27 of which were DTEK ELISA-positive for pork adulteration) revealed a wide range of pork content, with values as high as 48%.

Selective Procedure for Detection of Salmonellae from Poultry and Sausage Products.

The efficacy of lactose preenrichment and various selective enrichment and differential plating media were evaluated to determine the optimal procedure for detecting salmonellae from fresh chicken or frozen turkey, pork sausage, and cured chicken. Salmonellae were most frequently recovered from fresh poultry or pork sausage when samples were preenriched in lactose broth incubated at 35°C, selectively enriched in TT broth at 43°C, and streaked onto a new differential plating medium, modified lysine iron agar (MLIA/USDA). Enrichment of cured chicken in selenite brilliant green broth incubated at 43°C was more productive than in TT incubated at 43°C. When poultry and sausage samples were first preenriched at 35°C, selectively enriched at 43°C, and then streaked onto MLIA/USDA greater than 75% of all CFUs on the MLIA/USDA plates were typical of salmonellae. Different procedures are recommended for maximal recovery of salmonellae from fresh, frozen or cured poultry products.

Chloramphenicol-resistant Salmonella newport traced through hamburger to dairy farms. A major persisting source of human salmonellosis in California.

Animal-to-human transmission of drug-resistant salmonella and the role of antimicrobial use in food animals in the emergence of these bacteria are controversial subjects. Investigation of a 4.9-fold increase in Salmonella newport isolations from Californians in 1985 showed that 87 percent of the isolates had an unusual antimicrobial-resistance pattern (including chloramphenicol resistance) and a single, identical plasmid. Interviews of 45 patients and 89 matched controls in Los Angeles County showed that illness was associated with penicillin or tetracycline use during the month before onset (P less than 0.001) and with eating ground beef during the week before onset (P = 0.052). The epidemic strain was isolated from hamburger products eaten by cases, abattoirs where the animals from which the meat came were slaughtered, dairies that sent cows for slaughter on days when culture-positive products were processed, and ill dairy cows. Isolation of salmonella from beef carcasses in abattoirs correlated with the proportion of dead or moribund animals received for slaughter (r = 0.60, P less than 0.05). Isolation of chloramphenicol-resistant salmonella from dairy farms was associated with the use of chloramphenicol at those dairies. We conclude that food animals are a major source of antimicrobial-resistant salmonella infections in humans and that these infections are associated with antimicrobial use on farms.

Toronto General Hospital controlled trial data plasma exchange therapy in Guillain-Barré' syndrome.

The mechanism by which plasma exchange (PE) may benefit patients with acute Guillain-Barre' syndrome (AGBS) is unclear. It is possible, therefore, that the response of patients with AGBS is influenced by the choice of replacement solution (whole plasma vs. albumin or similar protein-containing solution). This report compares the outcome in 57 patients with AGBS treated at the Toronto General Hospital (TGH) using conventional therapy (27), PE with plasma replacement (15) and PE with albumin replacement (15). Fifteen patients (5 treated conventionally, 8 by PE with plasma replacement, 2 by PE with albumin replacement) were treated before the Baltimore coordinated multicenter trial. Forty-two patients (22 treated conventionally, 7 by PE with plasma replacement and 13 by PE with albumin replacement) were then entered at the TGH into the multicenter trial. The best outcome was observed in those patients (9) in whom PE was started within 14 days of the first neuropathic symptoms and plasma was used as replacement. The mean improvements in clinical grade in this subgroup of patients of 1.11 at four weeks after starting treatment and 1.71 at six weeks were significantly better than the corresponding improvements in the conventionally treated group of 0.35 (p less than 0.01) and 0.94 (p less than 0.05). The response of patients (9) exchanged within 14 days of onset, but replaced with albumin (grade change of 0.67 at four weeks and 0.86 at six weeks), was not significantly different from that of the conventionally treated patients. These data support the need for a randomized trial to compare PE using plasma replacement and PE using albumin replacement in patients with AGBS.(ABSTRACT TRUNCATED AT 250 WORDS)

Sampling Methods and Frozen Storage of Samples for Detection of Campylobacter jejuni on Freshly Processed Broiler Carcasses.

Swab, rinse and excision sampling methods are commonly used for detection of microorganisms on poultry carcasses. Swabbing has been the most frequently reported sampling method for Campylobacter jejuni on poultry. We evaluated the three methods for C. jejuni detection on freshly processed poultry in the following ways: (a) the interior and exterior surfaces of half of a carcass were each thoroughly rubbed with separate swabs which were combined in a test tube containing 2 ml of appropriate medium; (b) 25 g of skin and tissue samples from neck and abdominal opening cut areas were deposited in a stomacher bag with 5 ml of brucella broth (BB) and stomached for 2 min; and (c) half carcasses were shaken for 1 min with 100 ml BB in plastic bags. One drop of each sample was streaked for isolation on brucella agar containing 10% defibrinated sheep blood and Skirrow antibiotics. Isolates were identified by microscopy and appropriate cultural tests. All three sampling techniques were essentially equivalent for detection of C. jejuni on fresh carcasses. However, when samples were stored frozen for 7 to 10 d to simulate transport conditions from sampling locations to the laboratory, the incidence of detection was significantly reduced. Use of cryoprotective agents was an effective method to preserve swab samples during frozen storage.

Importance of glutamate synthase in glutamate synthesis by soybean cell suspension cultures.

The specific activities of glutamate synthase|EC 2.6.1.53, l-glutamine: alpha-ketoglutarate amino transferase (NADPH-oxidising)| and glutamine synthetase|EC 6.3.1.2, l-glutamate: ammonia ligase (ADP-forming)| extracted from soybean (Glycine max L.) cells grown in modified B5 medium were found to vary significantly in response to variations in the nitrogen content of the medium. The changes seen in specific activity levels could be correlated with similar patterns seen in the growth of the cells, in response to changes in the nitrogen content of the medium. By contrast, the specific activity of glutamate dehydrogenase|EC 1.4.1.2, l-glutamate: NAD(+) oxidoreductase (deaminating)|, was relatively low and invariant. Glutamate synthase was extracted from cells grown under optimal conditions, partially purified, and shown to have many properties in common with preparations of this enzyme extracted from other plant sources. Glutamate synthase was purified to homogeneity, using affinity chromatography on blue Sepharose.