Microbiological comparative study of isolates of Edwardsiella tarda isolated in different countries from fish and humans

It is difficult to use tissue culture assays to investigate adherence and other properties of Edwardsiella tarda because the organism is invasive and produces a potent hemolysin. We therefore relied on polymerase chain reaction (PCR) to determine the occurrence of genes for enterotoxins (LT-I, EAST-1), Shiga toxin (Stx-1, Stx-2), cytotoxic necrotizing factors (CNF-1, CNF-2), aerobactin, invasion plasmid of enteroinvasive Escherichia coli, EPEC adherence factor (EAF), intimin (Eae), enterohemolysin (EntHly) and hemolysin (Hly) in 53 isolates of E. tarda from humans and fish from several countries. All isolates were negative for all genes investigated by PCR. Adhesion to and invasion of HeLa cells were determined by using the unusually short incubation time of 1h or 30 min. All isolates adhered and invaded in these tests. Finally, a random amplified polymorphic DNA (RAPD) test distinguished, with a few exceptions, isolates of human and fish origin.     

Analysis of genetic relationship among Arbutus unedo L. genotypes using RAPD and SSR markers

The strawberry tree (Arbutus unedo L.) is an underutilized, drought tolerant, fire resistant species with a south western distribution in Europe, and with ecological and putative socio-economical impact in Portugal and Mediterranean countries. Our aim was to develop an appropriate set of molecular markers to enable genetic diversity to be assessed and to fingerprint Arbutus unedo genotypes for breeding and conservation purposes in Portugal. Twenty-seven trees from a broad geographic range were screened with 20 random amplified polymorphic DNA (RAPD primers) and 11 microsatellite markers (SSR). The RAPDs generated 124 bands, 57.3% of which were polymorphic, with an expected heterozygosity of 27%. We cross-amplified 11 SSR primers developed for Vaccinium spp., and 5 were found to be polymorphic in A. unedo, with 75% of expected heterozygosity, a number of alleles of 11.6, a null allele frequency of 7.6% and a polymorphic information content of 71%. Although the SSRs were more polymorphic and informative than the RAPDs, both markers displayed high genetic variability with the gathered data. No geographic pattern was observed in the genetic variation distribution based on both marker systems, and the lack of correlation between genetic and geographical matrices was confirmed by Mantel tests. Likely, no correlation was found between pairwise SSR and RAPD band-sharing matrices. These results and their implications on A. unedo breeding and conservation programs are discussed.     

Decline of Mediterranean oak trees and its association with Phytophthora cinnamomi: a review

Mortality events in cork and holm oaks have occurred in the Mediterranean basin since the beginning of the XX century, but severity of decline increased during the 1980s. By that time, the exotic soil borne pathogen Phytophthora cinnamomi was often recovered from declining stands and since then it has been considered the main factor associated with decline. This work analyses data concerning P. cinnamomi surveys in cork and holm oaks trees, pathogenicity tests carried out in controlled experiments, studies about the influence of site characteristics in tree decline and approaches to control the disease. Results of field surveys showed that the pathogen is widespread and pathogenicity tests suggested that host susceptibility to the pathogen is moderate when seedlings are in appropriate watering conditions, particularly cork oaks. Occurrence of decline is also associated with soil characteristics that interfere with root expansion and water retention. We assessed the relative importance of each factor involved in decline and revised the role of P. cinnamomi in cork and holm oak decline.     

Seed dispersal networks in an urban novel ecosystem

The conflict resulting from the expansion of human activities into natural habitats affects the structure and functioning of ecosystems. Similarly, the anthropogenic redistribution of many species all over the world affects the composition of biological communities, possibly altering their capacity to sustain key ecological functions, such as seed dispersal. Urban parks are extreme examples of such novel ecosystems resulting from the anthropogenic redistribution of species in a new ecological framework. Here, we describe the avian seed dispersal network in an urban recreational woodland in central Portugal (c.79 ha). Four quantitative seed dispersal networks were assembled by identifying intact seeds in the droppings of mist-netted birds throughout the year. Overall, 1,244 seeds were identified, representing 33 links between 15 plant species and 11 bird species. Most birds dispersed alien seeds, but these represented a small proportion of the overall network (20 % of the seeds and 13 % of the droppings). Blackcap (Sylvia atricapilla) was the main disperser of native and alien seeds in all seasons, particularly those of the invasive Phytolacca americana. Fleshy fruits were more abundant in summer, but were more consumed in winter, presumably when other foods were difficult to find. Our study suggests that even in a highly managed urban park, seed dispersal networks can be temporally complex and variable and that a network approach can be an important monitoring tool to detect the status of crucial ecosystem functions in rapidly changing habitats such as urban parks.     

The Effects of the Decaffeination of Coffee Samples on Platelet Aggregation in Hyperlipidemic Rats

The effect of coffee on cardiovascular diseases is still controversial. It is known that the process of decaffeination may influence the chemical constitution and, therefore, the biological effects of coffee. This study thus evaluated the effects of decaffeination on the levels of total phenols and chlorogenic acids in Coffea arabica L. samples, as well as the effects of ingesting both integral and decaffeinated coffee on the lipid profile and hemostatic and hematological parameters in normal and hyperlipidemic rats. Samples of integral and decaffeinated lyophilized coffee (Coffea arabica L., planted in Brazil) were used for chemical analysis (total phenols, chlorogenic acid and caffeine contents). For the bioassays, coffee beverages were prepared with non-lyophilized samples (10 % w/v) and were filtered and administered to animals by gavage (7.2 mL/kg/day) over 30 days. On the 31st day after beginning the treatment with coffee beverages, hyperlipidemia was induced to the animals by administering Triton WR-1339 (300 mg/kg body weight). On day 32, blood was taken to determine the lipid profile, platelet aggregation, prothrombin time, partially activated thromboplastin time and hemogram. The contents of both phenolic compounds and chlorogenic acid in the integral coffee beverage were significantly lower than those in the decaffeinated coffee beverage. The animals treated with Triton WR-1339 presented a mixed hyperlipidemia. Although the decaffeination process caused a relative increase in total phenols and chlorogenic acids, the coffee drinks were unable to change the lipid profile or the hemostatic and hematological parameters in the studied animals.     

Factors associated with seroprevalence of Neospora caninum in dairy cattle in southeastern Brazil

Neosporosis is an infectious disease caused by Neospora caninum, an obligate intracellular cyst-forming protozoan considered a major cause of miscarriage in dairy cattle in many parts of the world. This cross-sectional study evaluated the relationship between reproductive abnormalities and seropositivity to N. caninum in 1,204 dairy cows from 40 farms located in the state of Minas Gerais, southeastern Brazil. Producers were interviewed, and blood samples were collected to perform indirect immunofluorescence tests (IFAT 1:200). Association between reproductive abnormalities and seropositivity in cattle was evaluated with generalized estimating equations. The true herd-level seroprevalence of N. caninum was 95 % (83.3–99.1), while the individual-level true seroprevalence was 21.6 % (19.2–24.2). Several reproductive abnormalities were significantly associated with seropositivity to N. caninum: occurrence of repeated estrus (p?=?0.02; OR?=?3.84; 95 % CI?=?1.239–11.893), repeated miscarriages (p?=?0.001; OR?=?2.54; 95 % CI?=?1.423–5.402), and temporary anestrus (p?=?0.001; OR?=?3.44; 95 % CI?=?1.976–5.994). Furthermore, loose dogs (p?=?0.041; OR?=?2.20; 95 % CI?=?1.033–4.672) when fed raw meat (p?=?0.001; OR?=?1.91; 95 % CI?=?1.443–2.519) are risk factors for N. caninum infection. We observed that seropositivity to N. caninum in cattle increases risk of miscarriage by almost twice throughout the reproductive life of cows (p?=?0.004; OR?=?1.978; 95 % CI?=?1.249–3.131). Serologic evidence in this study indicates that N. caninum infection is widely distributed among dairy herds and significantly associated with reproductive disorders, especially miscarriage, repeated estrus, and temporary anestrus.     

Effect of three feeding management systems on some reproductive parameters of Scorpion mud turtles (Kinosternon scorpioides) in Brazil

Chelonians have been exploited since ancient times for their meat, eggs, fat, and offal, among other things. Among these animals, there is Scorpion mud turtle (Kinosternon scorpioides) which is an omnivorous turtle with semiaquatic habits and is widely consumed in Brazil. The aim of this study was to evaluate the influence of different feeding management systems on the reproductive performance of captive-bred Scorpion mud turtles, including egg quality and chemical composition. Study animals included 15 males and 48 females, divided into three groups. Animals were fed with either a commercial diet or a commercial diet supplemented with one of two different mixes (wet diet). All animals were fed at 1 % BW/day divided on Mondays, Wednesdays, and Fridays. Treatments were as follows: TR?=?100 % fish food (22 % CP); TRM1?=?70 % fish food?+?30 % mix 1 (bovine offal and fish); TRM2?=?70 % fish food?+?30 % mix 2 (bovine offal, fish, and shrimp). Samples were collected during 7 months, and eggs were identified, weighed, measured, and frozen for later analysis. Statistical analyses, including ANOVA, were performed using the program SAEG 9.0. Measurements taken from the nests were compared using the Tukey’s test (P?<?0.05). The different diets were associated with differential egg-laying performance and eggshell thickness. Animals fed with wet diets containing protein of animal origin displayed the best performance. Furthermore, nests with fewer eggs contained eggs of better external quality (e.g., greater length, width, and eggshell thickness). Finally, animals fed with only commercial feed produced eggs with lower saturated fatty acid content.