Characterization of a plasmid-encoded adhesin of an avian pathogenic Escherichia coli (APEC) strain isolated from a case of swollen head syndrome (SHS)

An avian pathogenic Escherichia coli (APEC) strain designated SHS4, isolated from a chicken with clinical signs of swollen head syndrome (SHS), adhered to but did not invade Hep-2 and tracheal epithelial cells. The PCR amplified fimA, csgA and tsh gene sequences. It produced Ia, Ib, E1, E3, K, and B colicins, but not colicin V and aerobactin. It harboured two plasmids of 60 and 98MDa and was resistant to streptomycin and tetracycline. Conjugation with a nalidixic acid (Na) resistant K-12 recipient strain (MS101) showed that the 98MDa plasmid did not transfer, whereas transfer of the 60MDa plasmid resulted in concomitant transfer of adhesion to Hep-2 and tracheal epithelial cells, production of the colicins Ia, E1, E3, and K, and the tsh-related DNA sequence. Transposon (TnphoA) mutagenesis of strain TR4 gave rise to strain Mut23, which lost its adhesive capacities, but was still able to express the same colicins as did strain TR4. PCR was able to amplify the tsh-related DNA sequence in this strain and a molecular probe based on transposon TnphoA indicated that the transposon was inserted in the 60MDa plasmid. Based on these results, we suggest that the 60MDa plasmid have adhesion genes, which may be responsible for the initial colonization of the upper respiratory tract of chickens.     

Virulence factors of avian pathogenic Escherichia coli isolated from broilers from the south of Brazil

Sixty-three Escherichia coli strains isolated from broilers with respiratory problems were examined for virulence factors, hemolysin synthesis ability, motility, hemagglutination capacity, operon pap presence, colicin production, and serum resistance. The capacity to hemagglutinate guinea pig erythrocytes was found in 53 (84.1%) of the samples, but only 30 (47.6%) agglutinated chicken erythrocytes. D-mannose-sensitive hemagglutination against guinea pig erythrocytes was found in 19 (30.2%) samples and against chicken erythrocytes, in 15 (23.8%) samples, whereas the D-mannose-resistant hemagglutination with guinea pig erythrocytes was found in 34 (54%) samples, and 13 of these (20.6%) showed this characteristic against chicken erythrocytes. Operon pap, P fimbria codifier, was detected in 26 samples in a total of 34 D-mannose-resistant samples. Colicin production was observed in 55 (87.3%) of the strains, and 41.8% presented V colicin production. Of the samples analyzed, 56 (88.9%) presented serum resistance, six (9.5%) were intermediate, and only one (1.6%) was sensitive to the action of the complement. The diversity of virulence profiles detected in the samples in this study explains in part the multifactorial characteristics of avian colibacillosis.     

First Portuguese isolate of Neospora caninum from an aborted fetus from a dairy herd with endemic neosporosis

Neospora caninum was isolated from the brain of an aborted 4-month-old fetus from a dairy cow herd with endemic neosporosis in Porto, Portugal. The fetal brain homogenate was inoculated interperitoneally first into outbred Swiss Webster mice given dexamethasone and then the peritoneal exudates from these mice was co-inoculated with mouse sarcoma cells in the peritoneal cavity of mice given dexamethasone. N. caninum tachyzoites were seen in peritoneal exudate of the second passage. Tachyzoites from the peritoneal exudate reacted positively with anti-N. caninum antibodies and not with anti-Toxoplasma gondii antibodies and contained N. caninum specific DNA. This Portuguese isolate of N. caninum has been successfully maintained in cell culture. The dam of the aborted fetus had an antibody titer of 1:10240 in the Neospora agglutination test (NAT). Antibodies to N. caninum were found in 76 of 106 cows from this herd in titers of 1:40 in 31, 1:80 in 22, > or =1:160 or more in 23 in the Neospora agglutination test. This is the first isolation of a viable N. caninum-like parasite from any host in Portugal.     

Development of Spinitectus inermis (Nematoda: Cystidicolidae), a parasite of eel,Anguilla anguilla,in Europe

The development of the nematode Spinitectus inermis (Zeder, 1800), a parasite of the stomach of eels, Anguilla anguilla (L.) in Europe, was experimentally studied. Mayfly nymphs Caenis macrura, Ecdyonurus dispar, Heptagenia sulphurea, Potamanthus luteus and Seratella ignita from Portugal and the Czech Republic were found to serve as experimental intermediate hosts. After ingestion of the nematode eggs by the mayfly nymphs, the toothed first-stage larvae were released and penetrated into the body cavity of the intermediate host. There they moulted twice (on day 4 and 6 post infection [p.i.] at water temperatures of 20-25 degrees C), attaining the third infective stage. The definitive host, A. anguilla, undoubtedly acquires infection by feeding on mayfly nymphs harbouring infective-stage larvae. In an experimentally infected eel, the fourth-stage larva undergoing the third moult was observed 28 days p.i. at water temperature of 20 degrees C. The larval stages, including moulting forms, are described and illustrated. The prepatent period of S. inermis is estimated to be about two months.     

Effects of species and sex on plasma hormone and metabolite concentrations in crossbred Brahman cattle and crossbred water buffalo

Buffalo meat is tasty and indistinguishable from beef, and has been described as being lean and low in cholesterol in comparison with beef. However, little is known about the plasma hormone and metabolic profiles related to fat metabolism in buffalo, and how their levels are affected by species and sex. In this study levels of plasma hormones and metabolites were compared between species and sexes. Ten growing crossbred Brahman cattle and ten growing crossbred buffaloes (five males and five females in each), with an average age of 22 months were used. The animals were fed on an experimental diet composed of corn silage, brewer's spent grain and a concentrate mixture at a 50:30:20 ratio on a dry matter basis during a fattening period of 6 months. Blood samples were collected every month during the experiment, and were analysed for plasma leptin, insulin, glucose, triglyceride (TG) and total cholesterol (TC) concentrations. Plasma leptin, insulin and TC concentrations increased during fattening and were significantly (P < 0.05) higher in cattle than in buffaloes. These concentrations in cattle showed a big difference between sexes in comparison with buffaloes, and were higher in females than in males. The plasma glucose and TG concentrations did not differ (P > 0.05) between cattle and buffaloes.

The crude fat content of longissimus muscle was significantly higher in cattle than in buffalo. Differences in body fat deposition seem to have an influence on plasma leptin, insulin and TC concentrations differently between cattle and buffaloes.     

Phylogenetic analysis based on ITS sequences and conditions affecting the type of conidial germination of Bipolaris oryzae

One taxonomic characteristic of Bipolaris species is the bipolar germination of conidia, but conidia of Bipolaris oryzae , the causal pathogen of brown spot in rice, are regularly observed to show intercalary germination, a characteristic of Drechslera species. The effect of selection, culture media and culture age on type of conidial germination was determined for three brown spot isolates from Cavinti, San Pablo and Palawan in the Philippines, obtained from infected leaves showing typical disease symptoms. Based on the analyses of their ITS1, ITS2 and 5·8S rDNA nucleotide sequences, the local isolates were clearly identified as B. oryzae . Selection for colonies of the three isolates derived from single conidia with either bipolar or intercalary germination had no effect on the number of spores showing bipolar germination in subsequent cultures. Germination on seven different culture media was tested; of these, rabbit food agar and water agar increased the percentage of bipolar germination of conidia, although this varied between isolates. Incubation of the cultures of all three isolates for longer periods prior to harvesting conidia increased the percentage of bipolar-germinating conidia from c . 40 to c . 90% with 5-day-old and 30-day-old cultures, respectively.     

Electro-acupuncture and Chinese herbs for treatment of cervical intervertebral disk disease in a dog

A non-ambulatory dog with tetraparesis following a pain episode that had evolved over 2 months was submitted for medical treatment and diagnosed with intervertebral disk disease at C3-C4 and dorsal extradural compression at C1-C2 and C3-C4 using myelography and computed tomography. The dog experienced ambulation recovery after 15 days of treatment with only electroacupuncture and Chinese herbal medicine, with marked improvement occurring after only 10 treatments. Six months of follow-up demonstrated that the dog was stable and had no recurrence of symptoms. Therefore, it was concluded that the combination of electroacupuncture and Chinese herbal medicine was responsible for motor rehabilitation.     

Molecular systematics in Chrysanthemum × grandiflorum (Ramat.) Kitamura

An attempt was made to understand the molecular systematics and genetic differences between 10 original chrysanthemum cultivars and 11 mutants. The similarity among the cultivars and mutants varied from 0.17 to 0.90 using RAPD analyses, a simple but efficient method to distinguish cultivars and to assess parentage. Two distinct groups were found. Two cultivars were present as a separate group showing differences from all other cultivars. Mutants with different flower colour could be identified at the molecular level using RAPD technique holding promise to identify unique genes as SCAR markers. A high genetic distance among the different chrysanthemums showed that there exists a possibility of introgressing new and novel genes from the chrysanthemum gene pool.