Comparison of viral RNA populations of pathogenically distinct isolates of Citrus tristeza virus : application to monitoring cross-protection

The population of sequence variants of Citrus tristeza virus (CTV) isolates of different geographic origins and pathogenicity properties was characterized by single-strand conformation polymorphism (SSCP) analysis of cDNA of the genes p18, p13, p20 and p23. The mild isolates analysed here usually yielded a SSCP profile with two DNA bands, suggestive of a predominant sequence variant, whereas the SSCP profile of the most virulent isolates contained more than two DNA bands, indicating that their viral populations are likely to be more complex. The set of SSCP profiles of the four genes allowed identification of individual isolates, but no profile characteristic of a geographic area or a biogroup was found. Sweet orange plants singly inoculated with a mild or with a severe isolate yielded the SSCP profile characteristic of each isolate, whereas the SSCP profile of plants successively inoculated with both isolates was a composite of the two individual profiles. The SSCP profile of plants singly inoculated remained constant, but the profile of doubly inoculated plants varied with time. Plants in which the SSCP profile of the severe isolate became predominant showed stem pitting, and those in which the predominant profile corresponded to the mild isolate remained symptomless. The results indicate that SSCP analysis can be used to study changes in RNA populations of doubly inoculated plants and to monitor cross-protection between mild and severe isolates.     

Post‐hatching development of the ornamental ‘Red Cherry Shrimp’ Neocaridina davidi (Bouvier, 1904) (Crustacea,Caridea, Atyidae) under laboratorial conditions

The ‘Red Cherry shrimp’, Neocaridina davidi is a small freshwater caridean shrimp living, originally, in various kinds of inland water bodies around Asian countries. This shrimp has reached several countries for ornamental use; however, basic information on the biology of the species is still scarce in the literature. Its early post embryonic development morphology has not yet been described. This paper focused on the production and the development of early post‐hatching stages of N. davidi, its male secondary sexual features, observation of the gonads and the presence of females with embryos. The larval development of N. davidi was almost suppressed as noted by the presence of relatively large‐sized eggs, first stage hatching as a decapodid; and the tail fans were present only from the 2nd post‐hatching stage. A biological important consequence of the presence of this Red Cherry shrimp species in the Neotropics is its potential release into nature, which could cause its rapid dispersion affecting populations of other indigenous caridean freshwater shrimps.     

Influence of Dietary Oil Sources on Muscle Composition and Plasma Lipoprotein Concentrations in Nile Tilapia,Oreochromis niloticus

To investigate the impact of different dietary lipid sources on fillet composition and lipid transport, we conducted a feeding trial and evaluated the proximate composition of muscle tissue, fatty acid profiles, total cholesterol (in muscle and plasma), triglycerides, and lipoprotein concentrations in Nile tilapia, Oreochromis niloticus. Five semi‐purified diets, containing different oils (soybean – SO, corn – CO, linseed – LO, fish – FO, and olive – OO), were supplied to tilapia for 160 d. Fish fed with LO and FO diets had a lower percentage of total lipids in muscle compared with the others (P < 0.05). The highest percentage of protein was found in fish fed with FO diet (P < 0.05). The muscle fatty acid profile was influenced differently by diets (P < 0.05). The group supplemented with SO and CO had a higher concentration of 18:2n‐6, whereas the fish fed with LO diet had a higher level of 18:3n‐3 and those that received the FO diet had more 22:6n‐3 in comparison with those supplemented with vegetable oils. Plasma lipid transport was also affected by the diets: the fish fed with FO diet had higher total cholesterol and high‐density lipoprotein and lower very‐low‐density lipoprotein concentrations (P < 0.05).     

Pericarp total protein profiles as molecular markers of tomato fruit quality traits in two segregating populations

The tomato fruit quality results of biochemical and physiological changes that occur during the ripening process. Although, the pericarp total protein profiles are less polymorphic than DNA-based markers the polymorphism in those could be directly associated with fruit quality traits. The aim of this work was to identify associations between polymorphic polypeptides from fruit pericarp at two ripening stages and fruit quality traits evaluated in two segregating populations of tomato. A cross between a normal ripening cultivar of Solanum lycopersicum (C, Caimanta) and a genotype carrying the nor (non ripening) gene (N) as well as a cross between Caimanta and a cherry type tomato of S. lycopersicum var. cerasiforme (Ce) showed genetic variance for several fruit quality traits such as fruit weight, shape, solids soluble content, acidity, color and fruit shelf life. The quantitative variations observed at phenotypic level had correspondence with the polymorphism detected in the protein profiles. Indeed, the polymophic polypeptides associated with quality fruit traits and fruit shelf life would be useful to assist tomato breeding programs as protein molecular markers.     

The value of molecular expression of KIT and KIT ligand analysed using real‐time polymerase chain reaction and immunohistochemistry as a prognostic indicator for canine cutaneous mast cell tumours

This study investigated the correlation between KIT gene expression determined by immunohistochemistry and real‐time polymerase chain reaction (RT‐PCR) and the rate of tumour recurrence and tumour‐related deaths in dogs affected with mast cell tumour (MCT). Kaplan–Meier curves were constructed to compare tumour recurrence and tumour‐related death between patients. The log‐rank test was used to check for significant differences between curves. KIT‐I, KIT‐II and KIT‐III staining patterns were observed in 9 (11.11%), 50 (61.73%) and 22 (27.16%) tumours, respectively. Tumour recurrence rates and tumour‐related deaths were not associated with KIT staining patterns (P = 0278, P > 0.05), KIT (P = 0.289, P > 0.05) or KIT ligand (P = 0.106, P > 0.05) gene expression. Despite the lack of association between KIT staining pattern and patient survival time, the results suggest a correlation between aberrant KIT localization and increased proliferative activity of MCTs. RT‐PCR seems to be a sensible method for quantitative detection of KIT gene expression in canine MCT, although expressions levels are not correlated with prognosis.     

Plasma biomarkers profile of female dogs with mammary carcinoma and its association with clinical and pathological features

The immunological biomarkers profiles were evaluated using Luminex as putative measures to monitor canine mammary carcinomas (MCs). Forty female dogs were categorized into benign mixed tumour (MC‐BMT = 28) and mammary carcinoma (MC=12). The ascendant biomarker signatures were used to compare the groups. For example, a higher frequency of MC‐BMT animals producing IL‐6, CXCL‐8 and CXCL‐10 was observed, whereas for the MC group IL‐2 and CXCL‐8 were detected. MC‐BMT animals without metastasis had an increase in the levels of IL‐2, CXCL‐8, CXCL‐10, IL‐6, TNF‐α, IL‐15 and a decrease in IL‐10 and CXCL‐8. MC‐BMT animals with metastasis showed only an increase in CXCL‐10 and a decrease in IL‐18. After comparing the ascendant signatures following the presence of metastasis in both groups, a higher frequency of dogs exhibiting IL‐10 production was observed. Pearson correlation (P = 0.0273) and receiver operating characteristic (ROC) curve analysis revealed that this pattern was associated with worse outcome and lower survival rates in MC animals.     

Characterization of symptoms of senescence and chilling injury on inflorescences of Heliconia bihai (L.) cv. Lobster Claw and cv. Halloween

Inadequate temperatures during the shipping and commercialization of cut tropical flowers may accelerate the senescence process and cause chilling injury, leading to symptoms that have not yet been described for Heliconia bihai. The aim of the present study was to evaluate physiological responses in cut inflorescences of H. bihai cv. Lobster Claw (LC) and cv. Halloween (HW) as well as symptoms of senescence and chilling injury. For such, changes in fresh weight, bract color (L*, a* and b*), percentage of absolute integrity (PAI) of cell membranes and leakage of potassium ions (LPI) were determined. The flowering stems were evaluated at five different intervals after harvest (0, 2, 4, 6 and 8 d). A refrigerated treatment (RT) with a temperature of 6.5 °C and 85% relative humidity was compared to a control treatment (CT) at room temperature of 24 °C and 66% relative humidity. Both cultivars stored at 6.5 °C exhibited dryness of bract tissue (symptom of senescence) and dark stains that became brownish and evolved to necrosis (symptom of chilling injury). The visual quality of inflorescences decreased with time in both cultivars maintained without refrigeration. The severity of chilling injury increased with the length of storage time in both cultivars. There was a significant reduction in the fresh weight of inflorescences in both treatments (RT and CT) and both cultivars (LC and HW). Bract color changed in both cultivars at 6.5 °C. There was no change in PAI throughout the evaluation period in the inflorescences stored at room temperature, whereas those stored at 6.5 °C for 6 and 8 d had lower PAI values. The inflorescences in the control treatment underwent no change in LPI values, whereas those stored under refrigeration had increased LPI values after the sixth day of storage. The physiological responses of cut Heliconia flowers were influenced by storage period and temperature, as demonstrated by visual symptoms of chilling injury and senescence.     

Outcomes of Heimlich valve drainage in dogs

Objective and design   Retrospective study of the outcomes of Heimlich valve drainage in dogs.
Procedure   Medical records of the past 3 years were retrospectively reviewed. Heimlich valve drainage was used in 34 dogs (median body weight 30 ± 5 kg): lobectomy (n = 15), pneumonectomy (n = 9), intrathoracic oesophageal surgery (n = 2), diaphragmatic hernia repair (n = 1), traumatic open pneumothorax (n = 2), bilobectomy (n = 2), ligation of the thoracic duct (n = 1), and chylothorax and pneumothorax (n = 1 each). Evacuation of air and/or fluid from the pleural cavity was performed with the Heimlich valve following thoracostomy tube insertion. During drainage, the dogs were closely monitored for possible respiratory failure. Termination of Heimlich valve drainage was controlled with underwater seal drainage and assessed with thoracic radiography.
Results   Negative intrathoracic pressure was provided in 29 dogs without any complications. Post pneumonectomy respiratory syncope and post lobectomy massive hemothorax, which did not originate from the Heimlich valve, were the only postoperative complications. Dysfunction of the valve diaphragm, open pneumothorax and intrathoracic localisation of an acute gastric dilatation–volvulus syndrome caused by a left-sided diaphragmatic hernia following pneumonectomy were the Heimlich valve drainage complications.
Conclusions   The Heimlich valve can be used as a continuous drainage device in dogs, but the complications reported here should be considered by veterinary practitioners.     

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.     

Phenotyping Castanea hybrids for Phytophthora cinnamomi resistance

Castanea sativa is susceptible to Phytophthora spp., a serious root pathogen causing ink disease, while C. crenata and C. mollissima show resistance to infection. Interspecific controlled crosses were established for introgression of resistance genes from the resistant species into the susceptible C. sativa, and two mapping populations were created. Phytophthora cinnamomi resistance of each progeny was evaluated by root and excised shoot inoculation tests. The number of days of survival after root inoculation was the best discriminator of resistance to P. cinnamomi while the percentage of shoots with internal lesions was the symptom most associated with survival. The lesion progression rate in the excised shoot inoculation test was strongly and negatively correlated with survival in the root inoculation test. The excised shoot inoculation test appears to be a reliable approach for screening the resistance of chestnut genotypes to P. cinnamomi. Strong genetic correlations were obtained between survival and ink disease symptoms and among symptoms, indicating that common or linked genes might influence resistance to P. cinnamomi. The most resistant genotypes selected from this study will be tested for other commercial variables, such as ease of vegetative propagation and stock–scion compatibility.