The role of toll-like receptors in the pathogenesis of Streptococcus suis

Streptococcus suis is an important agent of swine and human meningitis. Sequence type (ST) 7 emerged in China and was responsible for the human epidemic caused by S. suis in 2005. The virulence of S. suis ST7 is greater than the wild type pathogenic S. suis, ST1; however, the mechanisms for this increased pathogenicity are unknown. The aim of this study was to determine the role of different toll-like receptors (TLRs) involved in regulating the host response to the S. suis infection and to speculate on differing mechanisms used by ST7 strains to induce disease. Here we compared two ST7 strains isolated in the 2005 Sichuan outbreak to two ST1 strains. Our data show TLR2, 6 and 9 are involved in the recognition of heat-killed S. suis independent of the ST type. We found the TLR-dependent cytokine production differed between the two types of strains using whole cell lysate proteins. TLR6 played a greater role in cytokine production induced by the whole cell lysate proteins from the ST7 strain than in that induced by the ST1 strain lysates. The data suggest that mechanisms of inflammation induced by S. suis strains differ where this will be useful in designing efficient strategies in combating streptococcal toxic shock-like syndrome caused by the S. suis ST7 strains.     

Use of dimethylformamide to cryopreserve alpaca semen previously incubated with collagenase

The objective of this study was to evaluate the effect of collagenase and two final dimethylformamide (DMF) concentrations (4% and 7%) on alpaca frozen-thawed sperm quality. A total of 25 ejaculates from 5 alpaca were obtained using electroejaculation. Each individual ejaculate was evaluated and then diluted 4:1 in a solution of 1 mg/ml collagenase in HEPES-TALP medium and incubated for 4 min at 37°C. Subsequently, samples were diluted in TRIS-fructose-citric acid-egg yolk and cooled to 5°C. Then, each sample was divided in two aliquots and DMF at final concentration of 4% or 7% was added, equilibrated for 1 hr at 5°C and frozen over liquid nitrogen vapours. A Kruskal–Wallis test was used to evaluate the sperm morphometry, and Completely Random Block designs were used to analyse sperm motility, viability, membrane function and acrosome status. After collagenase incubation, none of the samples showed thread formation, and sperm parameters were preserved. Non-progressive motile sperm were higher (p < .05) in equilibrated samples (4% DMF: 31.8 ± 8.3% and 7% DMF: 36.3 ± 11.8%) compared to raw (10.1 ± 4.3%) and frozen-thawed semen (4% DMF: 9.7 ± 1.8% and 7% DMF: 7.5 ± 3.2%). Sperm membrane function, membrane integrity and intact acrosomes were higher (p < .05) in raw semen (40.1 ± 12.2%, 94.6 ± 3.2% and 91.3 ± 8.1%) compared to frozen-thawed samples (4% DMF: 19.8 ± 4.7%, 53.2 ± 2.7%, 65.7 ± 8.7% and 7% DMF: 20.4 ± 4.5%, 54.1 ± 1.4%, 64.6 ± 9.1%). Length of the sperm head was lower in frozen-thawed samples, being statistically different with 4% DMF compared to pre-freezing samples. The ratio between acrosome and head areas was greater (p < .05) in frozen-thawed samples. Incubation of raw alpaca semen with collagenase decreased the thread formation without affecting sperm quality. Frozen of collagenase treated alpaca semen with 4% or 7% DMF did not preserve the sperm parameters in thawed samples.     

Streptococcus suis infections in humans: the Chinese experience and the situation in North America

Infections caused by Streptococcus suis are considered a global problem in the swine industry. In this animal species, S. suis is associated with septicemia, meningitis, endocarditis, arthritis and, occasionally, other infections. Moreover, it is an agent of zoonosis that afflicts people in close contact with infected pigs or pork-derived products. Although sporadic cases of S. suis infection in humans have been reported, a large outbreak due to S. suis serotype 2 emerged in the summer of 2005 in Sichuan, China. A similar outbreak was observed in another Chinese province in 1998. Symptoms reported in these two outbreaks include high fever, malaise, nausea and vomiting, followed by nervous symptoms, subcutaneous hemorrhage, septic shock and coma in severe cases. The increased severity of S. suis infections in humans, such as a shorter incubation time, more rapid disease progression and higher rate of mortality, underscores the critical need to better understand the factors associated with pathogenesis of S. suis infection. From the 35 capsular serotypes currently known, serotype 2 is considered the most virulent and frequently isolated in both swine and humans. Here, we review the epidemiological, clinical and immunopathological features of S. suis infection in humans.     

Jaagsiekte sheep retrovirus found in milk macrophages but not in milk lymphocytes or mammary gland epithelia of naturally infected sheep

Jaagsiekte sheep retrovirus (JSRV) causes ovine pulmonary adenocarcinoma. JSRV can be transmitted via infected colostrum or milk, which contain somatic cells (SCs) harboring JSRV provirus. Nevertheless, the cell types involved in this form of transmission and the involvement of the mammary gland remain unknown. We separated adherent cells (macrophages and monocytes) by plastic adherence, and lymphocytes (CD4+ and CD8+ T cells, and B cells) by flow cytometry, from SCs in milk samples from 12 naturally infected, PCR blood test JSRV–positive, subclinical ewes. These cell populations were tested by PCR to detect JSRV provirus. The ewes were euthanized, and mammary gland samples were analyzed immunohistochemically to detect JSRV surface protein. We did not detect JSRV provirus in any milk lymphocyte population, but milk adherent cells were positive in 3 of 12 sheep, suggesting a potential major role of this population in the lactogenic transmission of JSRV. Immunohistochemistry did not reveal positive results in mammary epithelial cells, pointing to a lack of participation of the mammary gland in the biological cycle of JSRV and reducing the probability of excretion of free viral particles in colostrum or milk.     

Resistance of 57 greenhouse-grown accessions of Lycopersicon esculentum and three cultivars to Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae)

The aim of this study was to evaluate resistance to Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae) by antixenosis on 57 Lycopersicon esculentum Mill. accessions from the Horticultural Germplasm Bank (HGB) of Universidade Federal de Viçosa and by the three commercial cultivars (Santa Clara, Moneymaker and TOM-601) under greenhouse conditions. A randomized complete block design was used with three replications. Infestations with T. absoluta adults were performed weekly and the following characteristics were evaluated: number of small, large and total mines/leaf and % of leaves mined at days 60, 75 and 90 after planting. Low infestation occurred at days 60 and 75 but at day 90, infestation was sufficient to evaluate insect damage. Based on these data it was concluded that only accessions HGB-674 and HGB-1497 appeared to be the most promising. In addition, to determine possible chemical causes of resistance, hexane extracts were analyzed at day 90 by gas chromatography/mass spectrometry and the major peaks identified by a mass spectral database using similarity index. Nine hydrocarbons, viz., hexadecane, heptadecane, eicosane, tricosane, 2-methyltricosane, tetracosane, hexacosane, octacosane and triacontane were identified in the hexane extracts in many samples. Tricosane, tetracosane and hexacosane presented significant correlations with the leaves mined. Only tricosane presented a negative correlation with the number of small mines (r = −0.28), total number of mines (r = −0.27) and % of leaves mined (r = −0.22). However, tetracosane and hexacosane presented significant positive correlations (r = 0.25 and 0.24, respectively) with the % of leaves mined.     

Inheritance of Aluminum Tolerance and its Effects on Grain Yield and Grain Quality in Oats (Avena Sativa L.)

Summary Aluminum toxicity due to the cation Al⁺³ is a major factor limiting yields in acid soils. Wide genetic variability to aluminum tolerance is found in oat genotypes. The objectives of this study were to determine the number of genes controlling aluminum tolerance in oats and to verify if any detrimental effects were present of the aluminum tolerance genes on grain yield and grain quality in Al⁺³free soils. Aluminum tolerance was estimated as the average regrowth of the main root after exposure to toxic levels of Al⁺³ in a hydroponic solution under controlled conditions. The number of genes controlling that trait was estimated from the distribution of the average root regrowth frequencies in a population of 333 recombinant inbred lines (RIL's) in generations F_5:6 and F_5:7. The effects on grain yield and grain quality were assessed in a subpopulation of 162 RIL's chosen based on their aluminum tolerance response. Aluminum tolerance in the evaluated population was controlled by one dominant major gene with the tolerant genotypes carying Al _a Al _a and the sensitive ones al _a al _a alleles. No detrimental effects of the Al _a allele on grain yield or grain quality were detected.Part of the Master of Science dissertation of the first author