Seroprevalence and risk factors associated with neosporosis in sheep and dogs from farms

This study aimed to establish the seroprevalence and risk factors associated with neosporosis in sheep and dogs from rural properties. 1497 blood samples were collected from sheep and 42 from dogs that cohabited with sheep from 16 farms located in the central region of S?o Paulo State, Brazil. For the detection of N. caninum antibodies it was performed the indirect fluorescent antibody test (IFAT ≥ 25). For the epidemiological study it was applied a questionnaire for the owners or responsible from the sheep and dogs regarding informations related to neosporosis. The seroprevalence obtained out of the 1497 sheep sera tested was 8.0% (CI95%=6.7-9.2%) and out of the 42 dogs 4.8% (CI95%=0-7.2%). Variables statistically related to seropositivity for N. caninum in sheep were: dams well as water supply (P=0.0004; OR=2.15), presence of other domestic canids (P=0.0013; OR=2.38) and presence of reproductive problems (P=0.0031; OR=1.75).     

Coat protein-mediated protection against plum pox virus in herbaceous model plants and transformation of apricot and plum

Summary The expression of the viral coat protein gene in transgenic plants has been shown to induce tolerance against virus infection (Beachy et al., 1990). Transgenic plants ofNicotiana clevelandii andNicotiana benthamiana- herbaceous host plants for PPV - transformed withAgrobacterium strain LBA 4404 containing the plasmid pBinPPVm, regenerated on selection media containing kanamycin were tested for the expression of the PPV coat protein gene by ELISA and immuno western blot. After rooting and acclimatisation plants were tested for the protection against PPV Following the inoculation plants were investigated for symptom development and virus accumulation. Different lines were identified, according to the different reaction to the mechanical inoculation, ranging from a complete absence to a strong reduction of symptoms. There have not been many reports on transformation of trees in general, and in fruit trees particularly. It is obvious that the major obstacle is the regeneration of transformed plantlets. Attempts to improve crop plants by genetic engineering techniques will always depend very strongly on the availability of reliable protocols for transformation, selection and regeneration (Laimer et al., 1989, 1990). Different systems involving juvenile and adult plant material have been developed allowing the transfer of foreign genes into apricot and plum cultivars. We report the transformation and regeneration ofPrunus armeniaca andPrunus domestica plants withAgrobacterium tumefaciens strain LBA 4404 containing various binary plasmids, pBinGUSint, carrying the marker geneβ-glucuronidase (GUS) and pBinPPVm, carrying the coat protein gene of Plum Pox Virus (PPV), the causal agent of Sharka disease. The marker geneGUS was used for the optical evaluation of the efficiency of different transformation systems involving cotyledons of immature embryos as well as somatic embryos and leaf discs. The coat protein gene of PPV was used to introduce the coat protein mediated resistance against one of the most important pathogens of stone fruit trees in Europe and the whole Mediterranean area.     

Immunohistochemical Localization of Fibroblast Growth Factor‐2 in the Sheep Ovary and its Effects on Pre‐antral Follicle Apoptosis and Development In Vitro

Studies with sheep are important to improve our knowledge about the factors that control folliculogenesis in mammals and to explore possible physiological differences among species. The aims of this study were to characterize FGF‐2 protein expression in ovine ovaries and to verify the effect of FGF‐2 on the morphology, apoptosis and growth of ovine pre‐antral follicles cultured in vitro. After collection, one fragment of ovarian tissue was fixed for histological analysis and TUNEL analysis (fresh control). The remaining fragments were cultured for 7 days in control medium (α‐MEM⁺) alone or supplemented with FGF‐2 at different concentrations (1, 10, 50, 100 or 200 ng/ml). After culturing, ovarian tissue was destined to histology and TUNEL analysis, and oocyte and follicle diameters were measured. The immunostaining for FGF‐2 was observed in oocytes from primordial, primary and secondary follicles, as well as in granulosa cells of secondary and antral follicles. The percentage of normal follicles was similar among control medium, 1 and 10 ng/ml FGF‐2, and significantly higher than those observed in 50, 100 or 200 ng/ml FGF‐2. A significant increase in follicle diameter was observed when tissues were cultured in 10, 50, 100 or 200 ng/ml FGF‐2 compared with the fresh control and the other treatments. Similar results were observed for oocyte diameter in tissues cultured with 50, 100 or 200 ng/ml FGF‐2 (p < 0.05). However, the percentage of apoptotic cells only decreased (p < 0.05) in ovarian tissues cultured in 1 or 10 ng/ml FGF‐2 compared with the control medium and other FGF‐2 treatments. In conclusion, this study demonstrated the presence of FGF‐2 in ovine ovaries. Furthermore, 10 ng/ml FGF‐2 inhibits apoptosis and promotes ovine follicle growth. As the sheep ovary is more similar to that of humans, the culture system demonstrated in this work seems to be an appropriate tool for studies towards human folliculogenesis.     

A pseudo time‐series reveals the rapid recovery and high variability of benthic macroinvertebrate populations following catchment wildfire

1. The increasing risk of wildfire has focused attention on the timescale of the impact and recovery of river ecosystems and methods for their bioassessment.
2. An 18‐year pseudo time‐series was exploited to document patterns in benthic macroinvertebrate impact and recovery and evaluate the efficacy of alternative metrics to assess fire damage. Macroinvertebrates were surveyed by kick‐sampling and data were collected on river habitats. Details of river catchments and wildfire were collated as a GIS database.
3. Macroinvertebrate richness and abundance recovered rapidly, marked by a phase of dynamic increase, followed by relative stability (0–2 years and 3–18 years, respectively). Across sites, richness and abundance were best explained by time since fire.
4. A biotic index of general river quality was ineffective as an indicator of fire damage. While a metric of K‐selected taxa (Odonata richness) was generally indicative of fire‐affected assemblages, a contrasting metric of r‐selected taxa (percentage of chironomids, baetids, and simuliids) was not.
5. Ordination analysis revealed time as a significant determinant of community structure across sites; however, its overall statistical importance was eclipsed by habitat characteristics (water quality, shade, altitude, and latitude) that were associated with ecological variation across both recently affected sites and the putatively recovered communities.
6. These results highlight the stochastic processes – environmental and ecological – that frame the macroinvertebrate response to wildfire. This probabilistic context emphasizes the difficulties of developing indicator taxa for wildfire bioassessment and reinforces the importance of standardized survey protocols and the use of contrasting metrics in the assessment of wildfire impact on the ecological quality of rivers.

     

Distribution of oxidizable organic C fractions in soils under cacao agroforestry systems in Southern Bahia,Brazil

Agroforestry systems can play a major role in the sequestration of carbon (C) because of their higher input of organic material to the soil. The importance of organic carbon to the physical, chemical, and biological aspects of soil quality is well recognized. However, total organic carbon measurements might not be sensitive indicators of changes in soil quality. Adoption of procedures that can extract the more labile fraction preferentially might be a more useful approach for the characterization of soil organic carbon resulting from different soils. This study aimed to evaluate organic carbon (C) fractions distribution in different soil layers up to 50 cm depth in two soil orders under cacao (Theobroma cacao) agroforestry systems (AFS) in Bahia, Brazil. Soil samples were collected from four depth classes (0–5, 5–10, 10–30 and 30–50 cm) under two cacao agroforestry systems (30-year-old stands of cacao with Erythrina glauca, as shade trees) in Latosol and Cambisol, in Bahia, Brazil. The determination of oxidizable carbon by a modified Walkley–Black method was done to obtain four C fractions with different labile forms of C (fraction 1: labile fraction; fraction 2: moderate labile fraction; fraction 3: low labile fraction and fraction 4: recalcitrant fraction). Overall, at two cacao AFS, the C fractions generally declined with increase in soil depth. The C fractions 1 and 2 were 50% higher on upper layers (0–5 and 5–10 cm). More than 50% of organic C was found in more labile fraction (fraction 1) in all depths for both soils. High value of C fraction 1 (more labile C)-to-total organic C ratio was obtained (around 54–59%, on Latosol and Cambisol, respectively), indicating large input of organic matter in these soils.     

Use of a biophysical simulation model (DairyMod) to represent tall fescue pasture growth in Argentina

Tall fescue pastures have an increasing potential to be used worldwide. The purpose of this study was to test the ability and flexibility of the model DairyMod to represent herbage mass accumulation (HMA) of tall fescue pastures from Argentina under several environmental conditions, including different seasons, fertilizer nitrogen levels and irrigation. Species‐specific responses were obtained by customizing particular parameters (i.e. number of living leaves per tiller, leaf appearance interval, optimum plant N concentration and the response of leaf gross photosynthesis to temperature). Additionally, a simulation experiment to compare the model assumption that optimum N concentration stays constant through HMA with the application of a reference critical N dilution curve verified for temperate forage grasses (N% = 4·8 HMA^?0·32) was conducted. Application of DairyMod simulated with reasonable accuracy the HMA of tall fescue pastures under a wide range of climatic and management conditions; however, the model tended to underestimate HMA where pastures grew under high N availability. The use of a reference critical N curve substantially improved this bias, indicating that a further analysis on the N issue of the model is necessary. Results from this study provide support for further evaluations of the model under other scenarios and conditions. An upgrade of the model to improve simulation of N nutrition is suggested to enhance its performance to predict growth dynamics at high N availabilities as well as its value to address the effectiveness of N‐management practices.     

Antibody and blood leukocyte response in Rhipicephalus sanguineus (Latreille, 1806) tick-infested dogs and guinea pigs

The dog is considered to be the natural host of Rhipicephalus sanguineus and is unable to develop appreciable resistance even after repeated feedings. The guinea pig develops strong resistance after one infestation with adult ticks. Antibody (IgG) titres against tick salivary gland antigens (SGAs) and blood leukocyte numbers in dogs and guinea pigs undergoing experimental R. sanguineus tick infestations were measured to detect a possible correlation with susceptibility or resistance of hosts. Since infested dogs develop an immediate hypersensitivity reaction to R. sanguineus antigens, total and anti-R. sanguineus SGA IgE levels were also measured in this host species. IgG and IgE antibody levels were determined by an enzyme-linked immunosorbent assay (ELISA) along three consecutive infestations of both hosts. Most dogs and guinea pigs displayed low IgG levels against R. sanguineus SGAs, though marked differences in individual response were observed. Although dog's total serum IgE levels increased significantly after infestations, no change in the amount of anti-salivary gland IgE was detected. Total and differential blood cell counts were determined in dogs and guinea pigs during primary and secondary infestation. In dogs, a tertiary infestation and a subsequent higher infestation level were also evaluated. Infested dogs did not display any alteration in blood leukocyte counts throughout the experiment. Guinea pigs, on the other hand, developed a significant basophilia during primary infestation which increased further during secondary infestation. These data reveal similarities and differences in the reactions of resistant and non-resistant hosts to ticks. They contribute for the understanding of such host-parasite relationships and will hopefully aid in the development of immune control of ticks.     

Virulence genes and P fimbriae PapA subunit diversity in canine and feline uropathogenic Escherichia coli.

In this study, a total of 118 Escherichia coli strains isolated from dogs (93) and cats (25) with urinary tract infection (UTI) were tested in a multiplex polymerase chain reaction for the presence of adhesin-encoding genes (pap, sfa, and afa), hemolysin encoding genes (hly), cytotoxic necrotizing factor 1 (cnf1) and aerobactin (aer) genes. Virulence gene frequencies detected in those isolates which had been randomly collected (68 canine strains) were: 43% pap, 57% sfa, 1% afa, 44% hly, 41% cnf1 and 34% aer. These frequencies were much higher in the remaining 50 hemolytic strains of either cat or dog origin. Virulence factor associations in the 80 hemolytic strains studied revealed that 50/80 simultaneously had two adhesin genes (pap and sfa) and two cytotoxin genes (hly and cnf1), and 15/80 in addition had the aer gene. The major structural subunit and antigenic determinant of P fimbriae of uropathogenic E. coli is PapA. Polymorphism in this subunit was studied by an F antigen-specific papA allele polymerase chain reaction in 51 canine and 22 feline pap positive E. coli strains. The most prevalent canine papA alleles were F10 (39%), F15 (37%) and F12 (35%). In feline strains F15 (50%) was more frequent, other allele frequencies were F12 (45%), F14 and F10 (27%) and F16 (23%). Only nine canine and two feline strains were negative for one of the 11 serologically defined F types of P fimbriae. Three copies of the pap operon were found in 16/51 canine and 9/22 feline UTI E. coli pap positive strains. In this study, we show that a particular combination of virulence genes appears with high frequency in dog and cat urinary tract E. coli strains (pap, sfa, hly, and cnf1). In spite of the more frequent presence of F10, F12 and F15 papA alleles in this virulence gene combination, the occurrence of different papA alleles in strains where up to three copies of the pap operon are present accounts for the observed P fimbriae diversity.