Swine infection by Streptococcus suis: a retrospective study / Infecção em suínos por Streptococcus suis: estudo retrospectivo

The epidemic aspects of swine infections caused by Streptococcus suis were studied, focusing mainly on the occurrence of several serotypes. A total of 323 samples of S. suis were isolated from clinically ill animals, serotyped according to the co-agglutination procedure, and analyzed. The serotyping revealed that S. suis was present in several Brazilian states. The largest number was isolated from the states of Minas Gerais (62.5 percent), São Paulo (10.8 percent), and Paraná (9.3 percent). Serotype 2 was the most frequent (61.0 percent), followed by the serotypes 1, 3, 4, 7, and 8. The largest number of isolations was obtained from the brain (60.1 percent), followed by the lungs (10.4 percent). About 9.4 percent of the cases were due to septicemia.

Estudaram-se os aspectos epidêmicos das infecções de suínos causadas por Streptococcus suis, enfocando, principalmente, a ocorrência de diferentes sorotipos. Foram analisadas 323 amostras isoladas de animais clinicamente doentes, as quais foram sorotipadas de acordo com o procedimento de co-aglutinação. Foi verificado que S. suis está presente em vários estados brasileiros e o maior número de isolados originou-se dos estados de Minas Gerais (62,5 por cento), São Paulo (10,8 por cento) e Paraná (9,3 por cento). O sorotipo 2 foi o mais freqüente (61.0 por cento), seguido pelos sorotipos 1, 3, 4, 7 e 8. Os isolamentos foram obtidos principalmente de cérebro (60,1 por cento) e pulmões (10,4 por cento). Os casos de septicemia representaram 9,4 por cento.

Fermentation of starch by Klebsiella oxytoca p2, containing plasmids with alpha-amylase and pullulanase genes.

Klebsiella oxytoca P2(pC46), an ethanol-producing recombinant, has been evaluated in fermentation of maltose and starch. The maximum ethanol produced by P2(pC46) was 0.34 g ethanol/g maltose and 0.38, 0.40, or 0.36 g ethanol/g starch in fermentation of 1, 2, or 4% starch, representing 68, 71, and 64% the theoretical yield. The pC46 plasmid transformed to cells of K. oxytoca P2 reduced the ethanol production from maltose and starch. In fermentation of starch after its digestion at 60 degrees C for 24 h, in two-step fermentation, the time for maximum ethanol production was reduced to 12-24 h and the theoretical yield was around 90%. The increase in starch concentration resulted in lower alpha-amylase activity but in higher pullulanase activity. The high activity and thermostability of the amylolytic enzymes from this transformant suggest that it has a potential for amylolytic enzymes source.

Detection of Lawsonia intracellularis in faeces of swine from the main producing regions in Brazil.

Swine proliferative enteropathy is an enteric disease caused by Lawsonia intracellularis which affects animals between 6 and 20 weeks of age, causing diarrhea, anorexia, and poor growth. The presence of L. intracellularis was evaluated in the faecal samples of 636 swine from 75 randomly chosen herds in the main swine-producing regions of Brazil. The pathogen was detected by the polymerase chain reaction method (PCR) using L. intracellularis specific primers. A 319-bp DNA fragment specific for L. intracellularis was produced on amplification of DNA from the faeces of pigs with proliferative enteropathy. Equal amounts of DNA extracted from the faeces of animals from the same herd were pooled together and, once L. intracellularis was detected, the faecal material of each animal was analyzed separately. The incidence of L. intracellularis was 33.4% in the state of Santa Catarina, 29.4% in Paraná, 26.3% in Minas Gerais, 16.7% in Mato Grosso, and 7.1% in São Paulo. The presence of the pathogenic agent was detected in samples from 15 farms, representing a total incidence of 20%. Although 46 animals (7.2%) were shown to be infected, 11% did not present any symptoms of swine proliferative enteropathy. The use of PCR allowed the detection of L. intracellularis in swine farms and the evaluation of the incidence of proliferative enteropathy in different regions of Brazil.

Genetic diversity of the ectomycorrhizal fungus Pisolithus tinctorius based on RAPD-PCR analysis.

Twenty Pisolithus tinctorius isolates from different geographic locations and different hosts were characterized by the random amplified polymorphic DNA technique. Thirteen arbitrary primers generated 87 DNA fragments, all of them polymorphic. These data were used to calculate genetic distances among the isolates. The pairwise genetic distances ranged from 1 to 100%, with an average of 58.7%. Cluster analysis based on the amplified fragments grouped the isolates according to their host and geographical origins. Group I contained isolates collected in Brazil and group II those collected in the Northern Hemisphere. In addition to the diversity seen at the molecular level, the isolates also showed host specificity. Greenhouse experiments demonstrated that isolates from the Northern Hemisphere colonized mainly Pinus whereas isolates from Brazil colonized only Eucalyptus. The molecular data suggest that the Pisolithus tinctorius isolates analyzed belong to two distinct groups. The data also suggest new guidelines for future investigations on the taxonomy and systematic of this important fungus species. Furthermore, these results support future experiments aimed at the selection and development of improved isolates of P. tinctorius.

Production and regeneration of protoplasts from the mycorrhizal fungus Suillus granulatus.

Experiments were performed with the mycorrhizal fungus Suillus granulatus to define the parameters for production and regeneration of protoplasts. Protoplasts were released at frequencies between 1 and 3×10(7)/ml from mycelium 3 to 7 days old. The best osmotic stabilizer for protoplast release was MgSO4 (0.7 M). To optimize protoplast release and regeneration an enzyme (Novozym 234) concentration 1.7 mg/ml was chosen, with a digestion time of 1 to 2 h. Regenerated colonies formed mycorrhizae within 60 days after inoculation in Pinus caribaea var. hondurensis seedlings.

Fermentation of sweet whey by ethanologenic Escherichia coli.

Whey, an abundant byproduct of the dairy industry, contains large amounts of protein and lactose which could be used for fuel ethanol production. We have investigated a new organism as a candidate for such fermentations: recombinant Escherichia coli containing the genes encoding the ethanol pathway from Zymomonas mobilis. The highest level of ethanol achieved, 68 g/L, was produced after 108 hours in Luria broth containing 140 g lactose/L. Fermentations of lower lactose concentrations were completed more rapidly with approximately 88% of theoretical yields. Reconstituted sweet whey (60 g lactose/L)was fermented more slowly than lactose in Luria broth requiring 144 hours to produce 26 g ethanol/L. Supplementing sweet whey with a trace metal mix and ammonium sulfate reduced the required fermentation time to 72 hours and increased final ethanol concentration (28 g ethanol/L). By adding proteinases during fermentation, the requirement for ammonia was completely eliminated, and the rate of fermentation further improved (30 g ethanol/L after 48 hours). This latter increased in rate of ethanol production and ethanol yield are presumed to result from incorporation of amino acids released by hydrolysis of whey proteins. The fermentation of sweet whey by ethanologenic E. coil reduced the nonvolatile residue by approximately 70%. This should reduce biological oxygen demand and reduce the cost of waste treatment. Whey supplemented with trace metals and small amounts of proteinase may represent an economically attractive feedstock for the production of ethanol and other useful chemicals.

Resistance ofBradyrhizobium japonicum strains to selected antibiotics.

The majority of the 50Bradyrhizobium japonicum strains tested were resistant to ampicillin, kanamycin, streptomycin and tetracycline in concentrations below 100 µg/ml but resistant to chloramphenicol in concentrations equal to or above 100 µg/ml. Two strains had high levels of resistance to ampicillin and to streptomycin and six strains were very sensitive to several antibiotics.

Resistance ofBradyrhizobium japonicum strains to different fungicides and herbicides.

Of 50 strains ofB. japonicum, 64%, 44% and 54% were resistant to the fungicides captan, pentachloronitrobenzene and tetramethylthiuram disulphate, respectively, at concentrations above those recommended for treatment of soybeans. 100% and 84% of the strains were resistant to the herbicides trifluralin and metribuzin, respectively, also at concentrations above those recommended for soybeans. No strain was resistant to alachlor above 3.5 µg/ml but 40% were sensitive to 2.5 µg/ml, these being, respectively, the maximum and minimum concentrations recommended for soybeans.

Sobrevivencia de Salmonella em carne bovina moida armazenada em baixas tem peraturas. / Survival of Salmonella in ground beef under low storage temperature

Amostras de carne bovina moida, de cinco acougues da cidade de Vicosa, MG, foram armazenadas a 4 graus C por 14 dias, a 0 graus C e 18 graus C por 90 dias, para analise das populacoes de bacterias nao fermentadoras de lactose (Lac-) e determinacao da sobrevivencia e da resistencia de Salmonella a drogas antimicrobianas. As amostras, submetidas ao pre-enriquecimento em caldo lactosado, apresentaram-se com populacoes de bacterias Lac-superiores as amostras sem o pre-enriquecimento. Foi observado aumento nas populacoes de celulas Lac-, nos primeiros dias de armazenamento a 4graus C e a 0graus C. Apos 21 dias,houve reducao do numero de celulas Lac- nas amostras conservadas a 0graus C. Nas amostras a 18 graus C foi observada tendencia de reducao das populacoes de celulas Lacao longo do periodo de armazenamento. Celulas de Salmonella foram isoladas de amostras em todos os tempos e temperatura.o nivel de resistencia, aos varios antibioticos, das celulas de Salmonella, foi baixo