Microscale tracking of coral-vibrio interactions.

To improve our understanding of coral infection and disease, it is important to study host-pathogen interactions at relevant spatio-temporal scales. Here, we provide a dynamic microscopic view of the interaction between a coral pathogen, Vibrio coralliilyticus and its coral host Pocillopora damicornis. This was achieved using a microfluidics-based system facilitating control over flow, light and temperature conditions. Combined with time-resolved biochemical and microbial analyses of the system exudates, this approach provides novel insights into the early phases of a coral infection at unprecedented spatio-temporal resolution. We provide evidence that infection may occur through ingestion of the pathogen by the coral polyps, or following pathogen colonization of small tissue lesions on the coral surface. Pathogen ingestion invariably induced the release of pathogen-laden mucus from the gastrovascular cavity. Despite the high bacterial load used in our experiments, approximately one-third of coral fragments tested did not develop further symptoms. In the remaining two-thirds, mucus spewing was followed by the severing of calicoblastic connective tissues (coenosarc) and subsequently necrosis of most polyps. Despite extensive damage to symptomatic colonies, we frequently observed survival of individual polyps, often accompanied by polyp bail-out. Biochemical and microbial analyses of exudates over the course of symptomatic infections revealed that severing of the coenosarc was followed by an increase in matrix metaloprotease activity, and subsequent increase in both pathogen and total bacterial counts. Combined, these observations provide a detailed description of a coral infection, bringing us a step closer to elucidating the complex interactions underlying coral disease.

A coral-on-a-chip microfluidic platform enabling live-imaging microscopy of reef-building corals.

Coral reefs, and the unique ecosystems they support, are facing severe threats by human activities and climate change. Our understanding of these threats is hampered by the lack of robust approaches for studying the micro-scale interactions between corals and their environment. Here we present an experimental platform, coral-on-a-chip, combining micropropagation and microfluidics to allow direct microscopic study of live coral polyps. The small and transparent coral micropropagates are ideally suited for live-imaging microscopy, while the microfluidic platform facilitates long-term visualization under controlled environmental conditions. We demonstrate the usefulness of this approach by imaging coral micropropagates at previously unattainable spatio-temporal resolutions, providing new insights into several micro-scale processes including coral calcification, coral-pathogen interaction and the loss of algal symbionts (coral bleaching). Coral-on-a-chip thus provides a powerful method for studying coral physiology in vivo at the micro-scale, opening new vistas in coral biology.

Aetiology of Paediatric End-stage Renal Failure in Jordan: A Multicentre Study.

OBJECTIVE: The purpose of this study was to find out the aetiology of end-stage renal failure (ESRF) in children in Jordan. SUBJECTS AND METHODS: This was a multicentre retrospective study at five participating hospitals. Data collection included medical record review for age, gender, aetiology of ESRF, modality of renal replacement therapy (RRT) and outcome. End-stage renal failure was defined as estimated glomerular filtration rate < 15 mL/min/1.73m2. RESULTS: There were 275 children with ESRF: 131males and 144 females. The most common causes of ESRF in children were congenital anomalies of the kidney and urinary tract (CAKUT), 45.8%, heredofamilial disorders, 23.2% and glomerulopathies, 26.2%. Neurogenic bladder, reflux nephropathy and posterior urethral valve accounted for 16.8%, 12.7% and 4.0%, respectively. Amongst the heredofamilial disorders, primary oxalosis and cystic disease accounted for 8.0% and 7.2% of the aetiologies of ESRF, respectively. Focal segmental glomerulosclerosis was the most common histological type amongst the glomerulopathies (10.2%), followed by mesangiocapillary glomerulonephritis (4.7%) and chronic glomerulonephritis (3.0%). The aetiology was unknown in 4% of the cases. The modality of dialysis included isolated peritoneal dialysis (PD) in 30.9%, isolated haemodialysis (HD) in 49.1%, alternating peritoneal and haemodialysis in 9.1%, transplanted in 8.7% and conservative treatment in 1.8%. Death occurred in 57.3% of PD patients versus 34.4% in HD patients. CONCLUSIONS: This is the first report on the aetiology of ESRF in children in Jordan. The most common aetiologies of ESRF were CAKUT 45.8%, heredofamilial disorders 23.2% and glomerulopathies 22.9%.

Making the Patient Safety and Quality Improvement Act of 2005 work.

Potential reasons for failure to report medical errors include concerns about adverse publicity, fear of litigation and professional sanctions, the burden of reporting, uncertainty about what information is required to be reported, and lack of feedback. The Patient Safety Quality and Improvement Act of 2005 (PSQIA) was passed by the U.S. Congress as a response to these issues. Successful implementation of PSQIA requires changes such as moving toward an incentive-based system or providing continuing education programs for healthcare providers. In addition, the reform of residency programs will help in the implementation of PSQIA by future healthcare providers.

[Cysts from the first branchial cleft. Apropos of a case ]. / Kystes de la première fente branchiale. A propos d'un cas.

We describe a case of branchial cyst from the first branchial cleft in an 8-year-old patient, emphasizing the diagnostic difficulties of this type of lesion and the clinical aspects as well as the details of surgical treatment. The embryogenic origin and the anatomic relation with the facial nerve is recalled. Surgical treatment is indicated.

Intermittent fluconazole dosing in patients with onychomycosis: results of a pilot study.

BACKGROUND: Therapeutic limitations of griseofulvin in treating onychomycosis have led to a search for alternative antimycotic agents. An optimal dosing regimen for fluconazole has yet to be defined. OBJECTIVE: Our purpose was to evaluate the intermittent use of fluconazole (either once-weekly or alternate-day dosing) without concurrent nail avulsion in patients with moderate to severe onychomycosis. METHODS: Eleven patients with mycologically confirmed onychomycosis of the toenails or fingernails (43 infected nails) were treated with intermittent fluconazole until clinical cure was obtained. Eight patients received fluconazole 300 mg once weekly, one patient received 200 mg once weekly, and two patients received 100 or 200 mg of fluconazole every other day. Eight patients also used an adjunctive topical antimycotic preparation. RESULTS: All six patients with toenails involved (32 infected nails) were clinically cured after a mean treatment duration of 6 months, and all five patients with fingernails involved (11 infected nails) were cured after 3.7 months. There were no significant clinical or laboratory adverse events. CONCLUSION: Intermittent fluconazole, taken once weekly or on alternate days, is a well-tolerated and efficacious method to treat onychomycosis.

The superficial mycoses.

The various agents of the superficial mycoses have been recognized for more than a century as causes of mild diseases affecting humankind. Two of these, Malassezia furfur and Trichosporon beigelii, are ubiquitous organisms now known to be opportunistic pathogens in susceptible patient populations. The clinical manifestation, pathogenesis, and treatment of the common skin presentation of these and the other superficial mycoses are reviewed.

Detection of bovine coronavirus and type A rotavirus in neonatal calf diarrhea and winter dysentery of cattle in Quebec: evaluation of three diagnostic methods.

The use of direct electron microscopy, enzyme-linked immunosorbent assay, and protein A-gold immunoelectron microscopy for the identification of bovine coronavirus and type A rotavirus were examined. Two hundred and forty-nine samples from diarrheic calves and winter dysenteric cattle from seven geographic areas in Quebec were examined for the presence of viruses by direct electron microscopy of negatively stained preparations. In addition, all the samples were analyzed by enzyme-linked immunosorbent assay, and a random selection of 47 samples were also analyzed by protein A-gold immunoelectron microscopy. Thirty-nine percent of samples examined by direct electron microscopy contained viral particles; bovine coronavirus and type A rotavirus were the most common viruses involved. Overall agreement between any two of the methods used compared favorably with results obtained by others using similar methods. The presence of coronavirus and rotavirus in fecal samples obtained from neonatal calves and the presence of coronavirus in samples from winter dysenteric adult cattle suggested their etiological roles in the respective diseases. Furthermore, results from protein A-gold immunoelectron microscopy of coronavirus-like particles implied that a different coronavirus or some other viruses might be involved in these diseases. Finally, the efficiency of direct electron microscopy, enzyme-linked immunosorbent assay and protein A-gold immunoelectron microscopy as diagnostic tools is discussed.

Porcine respiratory coronavirus in Quebec: Serological studies using a competitive inhibition enzyme-linked immunosorbent assay.

Porcine respiratory coronavirus (PRCV) was identified for the first time in Quebec, using a blocking enzyme-linked immunosorbent assay (ELISA). Unlike the virus neutralization test (VNT), this ELISA was able to distinguish transmissible gastroenteritis virus (TGEV) from PRCV. Among the 15 seropositive fattening herds from group A, sera containing PRCV antibodies represented 74.8%, whereas those with TGEV antibodies represented only 7.2%. In group B, which consisted of 15 sow herds, nine herds expressed only PRCV-specific antibodies while the other herds had animals positive for TGEV-specific antibodies.

Antibacterial and antiviral activity of camel milk protective proteins.

Lysozyme (LZ), lactoferrin (LF), lactoperoxidase (LP), immunoglobulin G and secretory immunoglobulin A were extracted from camel milk. The activity of these protective proteins was assayed against Lactococcus lactis subsp. cremoris, Escherichia coli, Staphylococcus aureus, Salmonella typhimurium and rotavirus. Comparative activities of egg white LZ, bovine LZ and bovine LF are also presented. The antibacterial activity spectrum of camel milk LZ was similar to that of egg white LZ, and differed from bovine milk LZ. Bovine and camel milk LF antibacterial activity spectra were similar. The camel milk LP was bacteriostatic against the Gram-positive strains and was bactericidal against Gram-negative cultures. The immunoglobulins had little effect against the bacteria but high titres of antibodies against rotavirus were found in camel milk. The LP system was ineffective against rotavirus.

Differentiation of vaccine and field strains of bovine herpesvirus type 1 by restriction endonuclease analysis.

Bovine herpesvirus type 1 (BHV-1) DNA molecules obtained from a limited number of infected cells were cleaved with a variety of restriction endonucleases. By use of selected DNA fragments in Bgl 1, Pst 1, Pvu 1 and Pvu 11 restriction patterns, one reference, two vaccine and three wild-type strains of BHV-1 were distinguished from one another. The simplified DNA fingerprinting method described here should be most useful, not only to control the genetic stability of BHV-1 vaccines during production, but also to differentiate the vaccine strains from other isolates in clinical cases.

[The influence of previous infections on the development of experimental swine pleuropneumonia in specific pathogen free pigs]. / Influence d'infections préalables sur l'evolution de la pleuro-pneumonie porcine expérimentale chez des porcs exempts d'organismes pathogènes spécifiques.

This study was designed to determine in six-week old specific pathogen free pigs, the effect of previous experimental exposure to Mycoplasma hyopneumoniae and transmissible gastroenteritis virus on a challenge infection with Actinobacillus pleuropneumoniae. Pigs exposed simultaneously to M. hyopneumoniae and transmissible gastroenteritis virus appeared more resistant to challenge (one week later) with A. pleuropneumoniae. Four pigs out of a group of ten died following the challenge infection, compared to all ten pigs in the control group not submitted to previous infections. Clinical signs and lesions were also less severe in the previously infected group than in the control group. Pigs submitted to a single previous infection with M. hyopneumoniae only appeared to be less resistant to the challenge infection than pigs submitted to the dual previous infection with M. hyopneumoniae and the transmissible gastroenteritis virus. A correlation was found between the resistance of pigs to the challenge infection and their serum gammaglobulin levels.

Enzootic pneumonia in feeder pigs: Association with transmissible gastroenteritis virus infection.

Infection with transmissible gastroenteritis virus (TGEV) was present in some pigs on arrival at a feeder pig farm and was well established three weeks later. TGE infection preceded Mycoplasma hyopneumoniae infection, which was not detected until three weeks after arrival. Severe lesions of enzootic pneumonia were observed at the end of the fattening period.A trial was subsequently done in six-week-old-pigs in order to evaluate the potentiating effect of TGEV infection on experimental M. hyopneumoniae infection. The effects of mycoplasmal infection were aggravated when associated with TGEV infection as determined by the extent of pneumonic lesions observed two weeks later.

Immune response to an Actinobacillus pleuropneumoniae vaccine in swine.

Piglets vaccinated with an inactivated tetravalent vaccine (Pleurovac 4) against Actinobacillus pleuropneumoniae serotypes 1, 2, 5 and 7, produced circulating antibodies after a first intramuscular injection and showed an anamnestic reaction after a second. The rise in antibody levels in vaccinated piglets was statistically significant when compared with those of the control group. The administration of 1 or 2 mL of vaccine did not lead to significantly different antibody levels. The specificity of the immune response is demonstrated by an increase in titer to all four serotypes in pigs given the tetravalent vaccine, but an increase in titer to only two serotypes in pigs given a bivalent vaccine (Pleurovac).

Genotypic characterization of type-2 variants of canine adenovirus.

Two canine adenovirus (CAV) isolates, apparently distinct from type-1 (Utrecht) and type-2 (Toronto A26/61) reference strains in their biochemical and/or immunologic properties, were submitted to DNA-restriction endonuclease analysis. Both isolates, designated IAF-81-2116 and IAF-75-95, appeared as genotypic variants of CAV-2. Isolate IAF-81-2116 was recovered from the intestine of a young pup with diarrheal disease. Seemingly, relatively small changes in the original CAV-2 DNA sequence allowed the virus to replicate at an unusual site in the dog.

Endonucléases de restriction: isolement et identification d'une souche d'adénovirus canin de type 1.

For the first time in Quebec, a type 1 canine adenovirus was isolated in cell culture and typed by restriction endonuclease analysis. This virus originated from the internal organs of a young dog killed by a severe respiratory disease without showing any sign of hepatitis.

Interspecific differences between the DNA restriction profiles of canine adenoviruses.

Identification of canine adenovirus-1 ( CAV -1) and canine adenovirus-2 ( CAV -2) strains was done by electrophoresis of restriction endonuclease-fragmented viral DNA. Results obtained with this sensitive and reproducible technique clearly show that CAV -2 is not a variant of CAV -1 but a distinct virus.

Unambiguous typing of canine adenovirus isolates by deoxyribonucleic acid restriction-endonuclease analysis.

Viral deoxyribonucleic acid extracted from a limited number of cells infected with canine adenovirus type 1 or type 2 was cleaved with several restriction endonucleases. Agarose gel electrophoresis of the limit digests showed stable differences between the canine adenovirus type 1 and type 2 cleavage patterns. Rapid and accurate typing of large numbers of clinical isolates may thus be done by deoxyribonucleic acid restriction-endonuclease analysis.