Potential Utilization of Green Tide-Forming Macroalgae from Patos Lagoon,Rio Grande-RS,Brazil

ABSTRACT

This study was conducted to evaluate the biochemical and nutritional characteristics of naturally occurring estuarine macroalgae in the Patos Lagoon (Rio Grande, Brazil). The study of these macroalgae, dominated by Ulva and Cladophora species, contributes to current research on the potential utilization of green tide-forming algae. Although high levels of carotenoids (378 ± 0.02 µg g^?1) and antioxidant activity (75.0% for DPPH) were found in the naturally occurring macroalgae, these macroalgae cannot be used in the food industry due to the presence of toxic levels of arsenic (exceeding the thresholds of both French and Brazilian legislation for food). Regarding the potential of the studied macroalgae as a biofuel source, the low lipid and very high carbohydrate contents suggest a possible use in the fermentation of ethanol, butanol, or methane.     

Effect of Ischemia and Reperfusion on Neutrophil Accumulation in Equine Microvascular Tissue Flaps

Objective— To investigate neutrophil accumulation after ischemia and reperfusion (IR) in microvascular tissue flaps in horses.
Study Design— Randomized controlled experiment.
Sample Population— A total of 8 horses between 1 and 10 years of age, 4 of each sex.
Methods— Control and experimental myocutaneous island flaps based on the superficial branch of the deep circumflex iliac vessels were dissected on each horse. Atraumatic vascular clamps were applied to the pedicle of the experimental flap for 90 minutes and then removed to allow reperfusion. Based on the assumption that rapid infiltration of neutrophils into affected tissues is a hallmark of IR injury, radiolabeled autogenous leukocytes were used to indirectly quantify neutrophil accumulation in flap tissues. Labeled leukocytes were administered through a jugular catheter 30 minutes before flap reperfusion. Biopsies were collected from each flap over a 6 hour postischemia time period; in group 1 (  n = 4  ) from 0 to 6 hours postischemia, and in group 2 (  n = 4  ) from 24 to 30 hours postischemia. Biopsies were examined scintigraphically and histologically for evidence of neutrophil infiltration.
Results— All control flaps survived and 6 of 8 experimental flaps survived. There was no significant evidence of acute neutrophil infiltration into flap tissues after reperfusion in either group.
Conclusions— The results of this study suggest that equine myocutaneous flap tissues can survive a 90-minute ischemic period and reperfusion. No significant evidence of the occurrence of IR injury in flap tissues was found.
Clinical Relevance— The reasons for the previously reported failures of equine free tissue transfer remain uncertain, but they do not appear to be caused by neutrophil mediated injury associated with ischemia and reperfusion.     

Prevalence of Ehrlichia canis infection in thrombocytopenic dogs from Rio de Janeiro, Brazil

BACKGROUND: Infection with Ehrlichia canis causes a highly variable, multisystemic disease in dogs. Nevertheless, many clinicians in Rio de Janeiro, Brazil, use the presence of only thrombocytopenia to make a presumptive diagnosis of E canis infection. OBJECTIVE: The objective of this study was to determine the prevalence of E canis in thrombocytopenic dogs from Rio de Janeiro, Brazil, using polymerase chain reaction (PCR). METHODS: Following DNA extraction of whole blood samples from 226 dogs, PCR assays were done using primers for rickettsial DNA (including Ehrlichia spp, Anaplasma platys and A phagocytophilum) and using E canis-specific primers (16S rRNA gene). Dogs were grouped as thrombocytopenic and nonthrombocytopenic based on platelet counts. The null hypothesis that there was no difference in the prevalence of E canis in these groups was rejected at P<.05. RESULTS: Thirty-six (32.1%) of the thrombocytopenic dogs and 4 (3.5%) of the nonthrombocytopenic dogs were positive for rickettsial gene sequences (P<.0001). Further, 30 (26.8%) of thrombocytopenic dogs and 4 (3.5%) nonthrombocytopenic dogs were positive for E canis-specific gene sequences (P<.0001). CONCLUSIONS: Although the prevalence of E canis infection was higher in thrombocytopenic dogs, less than one third of these dogs had demonstrable E canis infection. Thus, thrombocytopenia is not specific for the detection of E canis infection and should not be used solely to establish a diagnosis of canine ehrlichiosis, even in a geographic area with relatively high disease prevalence.     

Damage by Homalinotus depressus in commercial coconut palm crops in the Amazonian region of Brazil

The coconut palm (Cocos nucifera L., Arecaceae) has great economic and social importance for many industrial and non-industrial products. Pests can reduce its productivity and cause its death. In 2005, larvae and adults of a Curculionidae were observed damaging the floral stalk of coconut palms in commercial plantations in the municipality of Moju, Pará State, in the Brazilian Amazon. Insects were identified as the black coconut bunch weevil, Homalinotus depressus (L.) (Coleoptera: Curculionidae: Cholini). This is the first report of this pest damaging coconut palms in Brazil.     

Effects of Topical Application of Sunflower-Seed Oil on Experimentally Induced Wounds in Horses

The objectives were to evaluate clinical and histopathological aspects of topical application of sunflower-seed oil (Helianthus annuus) on the healing process of experimentally induced wounds in lumbar and metacarpal areas of healthy horses. Six adult horses were used. Eight wounds were surgically produced on each horse: two 6.25-cm² wounds cranial to the sacrum on each side of the lumbar region and two 2.89-cm² wounds close to the proximal epiphysis of the metacarpus on the lateral aspect of each forelimb. Left side was used for macroscopic observations and right side for histopathological analysis. The experimental lesions were treated daily with sunflower-seed oil, whereas saline solution was used in control lesions. Macroscopic and histopathological analyses were performed on tissue harvested at 3, 7, 14, and 21 days. Complete healing time for all wounds was recorded. For lumbar region’s wounds, a contraction of 90.78% was recorded for those treated with oil and of 79.27% for control wounds after 21 days of treatment. For metacarpal region’s wounds, a contraction of 47.63% was recorded for wounds treated with oil and of 30.21% for control wounds. Wounds in the sunflower-seed oil treatment group had an elevation of polymorphonuclear cells, a newly formed vascular bed during the inflammatory phase, and a better alignment of collagen fibers during the remodeling phase. In conclusion, topical application of sunflower-seed oil was beneficial in the healing process of experimentally induced skin wounds in horses, with best results for treatment of lumbar wounds, making it a therapeutic option in equine wound healing.     

Effects of Short-term Hyper- and Hypoprolactinaemia on Hormones of the Pituitary, Gonad and -Thyroid Axis and on Semen Quality in Male Beagles

Effects of a short-term hyper- and hypoprolactinaemia on serum concentrations of LH, testosterone and semen quality in six male Beagles were investigated. Blood samples were collected at 3-day intervals for 12 weeks. The time span was divided into five 3-week periods: pre-treatment, metoclopramide (MCP) treatment (0.2 mg/kg orally three times daily), cabergoline (CAB) treatment (5 μg/kg orally once daily), post-treatment 1 and post-treatment 2. In the latter, only semen characteristics were evaluated. Semen parameters were analyzed once per week during the whole 15-week investigation time. At the end of each period, the effects of a single intravenous injection of thyrotropin-releasing hormone (TRH; 10 μg/kg) on the secretion of prolactin (PRL), LH, testosterone, thyroid-stimulating hormone and thyroxine (T4) were investigated. Pre-treatment serum PRL concentration increased under MCP (p < 0.05), followed by a decrease under CAB administration (p < 0.05). Luteinizing hormone and testosterone concentrations were not affected. Except for straight-line sperm velocity, semen quality did not differ between collection periods. A single iv TRH injection induced a significant PRL increase at 20 min in all experimental periods except during CAB treatment. Luteinizing hormone and testosterone did not show clear TRH-related changes. Basic T4 levels were significantly reduced after CAB treatment (p < 0.05). The results of the present study demonstrate that MCP-induced short-term hyperprolactinaemia in male beagles does not seriously affect the hypothalamo-pituitary axis and semen quality.     

Relationships Between Angular Leaf Spot, Healthy Leaf Area, Effective Leaf Area and Yield of Phaseolus vulgaris

Three field experiments were carried out with the bean cultivar Carioca Comum to investigate the relationships among visual and virtual severity of angular leaf spot (caused by Phaeoisariopsis griseola), area under visual and virtual disease progress curves (AUDPC), healthy leaf area index on any given day (HLAI), healthy leaf area duration (HAD), healthy leaf area absorption (HAA), effective leaf area duration (ELAD), effective leaf area absorption (ELAA) and yield of Phaseolus vulgaris. To obtain a wide range of disease severities, the plots were sprayed with fungicide at different stages of plant growth (before, during and after flowering). Visual and virtual severity and AUDPC showed no significant correlation with yield. However, HAD, HAA, ELAD and ELAA were significantly correlated with yield. Variables that considered the effective leaf area (ELAD and ELAA) provided similar or better coefficients of determination (R²) than those that considered the remaining green leaf area only (HAD and HAA). Single-point models with HLAI, effective leaf area index (ELAI), intercepted radiation by healthy leaf area (HRI) and intercepted radiation by effective leaf area (EHRI) to estimate yield at various times during the crop season were developed. The slope of the relationship between yield and HLAI, ELAI, HRI and EHRI proved to be stable, regardless of planting date and bean growth stage (from R6 to R8).     

Following the dynamics of strobilurin resistance in Blumeria graminis f.sp. tritici using quantitative allele-specific real-time PCR measurements with the fluorescent dye SYBR Green I

Strobilurin-resistant isolates of Blumeria ( Erysiphe ) graminis f.sp. tritici , the cause of wheat powdery mildew, were more than 10-fold less sensitive to azoxystrobin than sensitive isolates. In all resistant isolates, a mutation resulting in the replacement of a glycine by an alanine residue at codon 143 (G143A) in the mitochondrial cytochrome b gene was found. Allele-specific primers were designed to detect this point mutation in infected wheat leaves. Using quantitative fluorescent allele-specific real-time polymerase chain reaction (PCR) measurements, strobilurin-resistant A143 alleles could be detected amongst strobilurin-sensitive G143 alleles at a frequency of at least 1 in 10 000, depending on the amount of target and nontarget DNA. Most isolates tested were dominant homoplasmic for either the A143 or G143 allele, although mixed populations of alleles could be detected in some isolates. In some of these isolates, strobilurin resistance was not always stable when they were maintained for many generations in the absence of selection. The allele-specific real-time PCR assay was also used to follow the dynamics of A143 alleles in field populations of B . graminis f.sp. tritici before and after application of fungicides. As expected, the A143 allele frequency only increased under selection pressure from a strobilurin fungicide. After three sprays of azoxystrobin, a pronounced selection for the strobilurin-resistant allele, with an increase in average frequency from 2·2 to 58%, was measured. The use of quantitative real-time PCR diagnostics for early detection of fungicide resistance genes at low frequency, coupled with risk evaluation, will be invaluable for further resistance risk assessment and validation of antiresistance strategies.     

Assessment of primers designed for the subspecies-specific discrimination among Babesia canis canis, Babesia canis vogeli and Babesia canis rossi by PCR assay

Canine babesiosis is an infectious disease caused by either Babesia gibsoni or Babesia canis protozoans. The latter is also classified under three different phylogenetic groups, referred to as subspecies B. canis canis, B. canis vogeli and B. canis rossi. The objective of the present study was to validate and standardize a PCR assay to discriminate the organisms at the subspecies level. First, the reference sequences of the 18S rRNA, 5.8S rRNA and 28S rRNA genes, including the internal transcribed spacer 1 (ITS1) and 2 (ITS2) of the most common species and subspecies of the genus Babesia were retrieved from the GenBank database. Subspecies-specific primers (BAB3, BAB4 and BAB5) and one genus-specific primer were designed from the alignment of the sequences. The PCR assays were evaluated in three different combinations of primer pairs in order to assure complete specificity for each reaction. The results of the tests had demonstrated effectiveness of the novel primer pairs BAB1/BAB3, BAB1/BAB4 and BAB1/BAB5 for the amplification of the subspecies-specific target fragments of 746 bp (B. c. canis), 546 bp (B. c. vogeli) and 342 bp (B. c. rossi) by PCR. The original enzymatic amplification assays with novel primers reported in this paper were confirmed to be a reliable tool for the specific discrimination among B. canis subspecies by single-step PCR assays.