Xylella fastidiosa is a xylem-dwelling, insect-transmitted, gamma-proteobacterium that causes diseases in many plants, including grapevine, citrus, periwinkle, almond, oleander, and coffee. X. fastidiosa has an unusually broad host range, has an extensive geographical distribution throughout the American continent, and induces diverse disease phenotypes. Previous molecular analyses indicated three distinct groups of X. fastidiosa isolates that were expected to be genetically divergent. Here we report the genome sequence of X. fastidiosa (Temecula strain), isolated from a naturally infected grapevine with Pierce's disease (PD) in a wine-grapegrowing region of California. Comparative analyses with a previously sequenced X. fastidiosa strain responsible for citrus variegated chlorosis (CVC) revealed that 98% of the PD X. fastidiosa Temecula genes are shared with the CVC X. fastidiosa strain 9a5c genes. Furthermore, the average amino acid identity of the open reading frames in the strains is 95.7%. Genomic differences are limited to phage-associated chromosomal rearrangements and deletions that also account for the strain-specific genes present in each genome. Genomic islands, one in each genome, were identified, and their presence in other X. fastidiosa strains was analyzed. We conclude that these two organisms have identical metabolic functions and are likely to use a common set of genes in plant colonization and pathogenesis, permitting convergence of functional genomic strategies.Different microorganisms are able to survive in and to colonize plant water-conductive vessels (xylem). The result of this association is either beneficial or detrimental to the plant host.Of the latter, an example is the association of Xylella fastidiosa (38) with diverse plant hosts. X. fastidiosa is a fastidious, insecttransmitted, xylem-inhabiting bacterium known to cause several economically important diseases of both monocotyledonous and dicotyledonous plants (14,17,29). These diseases include Pierce's disease (PD) of grapevine and citrus variegated chlorosis (CVC), which have rather distinct symptoms and geographical distributions.PD, caused by certain strains of X. fastidiosa, is characterized by wilted, shriveled, raisin-like fruit and scorched leaves that detach, leaving bare petioles attached to the canes (37). The bark of affected canes may lignify or mature irregularly, leaving

This study was carried out to evaluate the performance of Brazilian Somalis sheep to natural infections by gastrointestinal nematodes. During 98 days, 75 weaned sheep, initially 3-4 months old, were kept on the same pasture and evaluated. Fecal and blood samples were collected for parasitological and hematological exams. After this period, the eight most resistant and the eight most susceptible animals were selected based on their individual averages of nematode fecal egg counts and were slaughtered for worm burden determination and nematodes identification. Abomasum and abomasum lymph nodes were also recovered for gene expression analysis. The animals selected as resistant had lower fecal egg counts during experimental period and smaller worm burdens than the susceptible ones (P < 0.05). The genus Haemonchus, followed by Trischostrongylus and Oesophagostomum, were identified in composite cultures. Haemonchus contortus was the specie identified in the abomasum. Packed cell volume and total plasma protein means were higher in the resistant group (27.2% and 6.1 g/dL) than in the susceptible one (22.5% and 5.3 g/dL), respectively. Regarding cytokine gene expression, IL-4 (P < 0.05) was up-regulated in the abomasum of resistant animals and TNF-α (P < 0.03) and IFN-γ (P < 0.03) in susceptible ones. In abomasum lymph nodes, IL-4 (P < 0.04) and IL-13 (P < 0.05) were up-regulated in the resistant animals and IFN-γ in the susceptible one (P < 0.01). This work provides further evidence that, within a given animal breed, individuals have different responses when infected by gastrointestinal nematodes. Resistant animals who responded more quickly and efficiently to these infections activated a TH2-type response.

Genetic differences in susceptibility to ticks (Rhipicephalus (Boophilus) microplus) are considerable in bovines. Here, mapping, association and gene expression approaches were employed to further advance our understanding of the molecular basis of tick resistance. A B. taurus x B. indicus F2 population was developed by Embrapa and 382 individuals were measured for parasitic load. Scanning of all chromosomes is in progress. Quantitative trait loci (QTL) for tick load were mapped to chromosomes 4, 5, 7, 10, 14, 18 and 23 out of the 20 chromosomes scanned and were dependent on the season in which the phenotype was scored. In the candidate gene approach, females from the genetic groups Nelore (NE--184), Canchim x Nelore (CN--153), Aberdeen Angus x Nelore (AN--123) and Simmental x Nelore (SN--120) were evaluated under natural infestation. Microsatellite markers close to the genes for interleukin 2 (IL2), interleukin 4 (IL4) and interferon gamma (IFNG) were analysed. Tick counts were associated with the marker for interleukin 4 (P < 0.05) in three genetic groups. Differences in cytokine mRNA levels of naive versus infested Nelore calves as well as between resistant versus susceptible cows from NE, CN and AN genetic groups were also investigated. Comparison of cytokines from infested and naïve animals showed downregulation of IL2. When resistant cows were compared to susceptible animals, IL8 was downregulated. These results reinforce the multiloci nature of tick resistance and the need to consider QTL and environment interactions.

This study aimed to evaluate the expression of a subset of cytokine genes in response to Haemonchus placei infections in Nelore cattle presenting different degrees of resistance to natural infections. One hundred weaned bulls, initially 11-12 months old, were evaluated and kept on the same pasture. Faecal and blood samples were collected for parasitological and immunological assays. The seven most resistant and the eight most susceptible animals were selected based on nematode faecal egg counts (FEC) and worm burden. Serum was collected to measure antibody titres, and abomasum and abomasal lymph node tissue samples were collected to analyse the expression of a subset of cytokine genes (IL-2, IL-4, IL-8, IL-12p35, IL-13, TNF-alpha, IFN-gamma, MCP-1, MCP-2, MUC-1) using real-time RT-PCR. Mast cells, eosinophils and globule leukocytes in the abomasal mucosa were enumerated, and IgA levels in the mucus were assessed. Gene expression analysis in the abomasal tissue indicated that IL-4 and IL-13 (TH2 cytokines) were up-regulated in the resistant group, whereas TNF-alpha (TH1/TH2 cytokine) was up-regulated in the susceptible group. In abomasal lymph nodes, IL-4 and IFN-gamma were up-regulated in the resistant and susceptible groups, respectively. In the resistant group, serum IgG1 levels were higher against antigens of H. placei infective larvae on days 14, 42, 70 and 84 and against antigens of H. placei adults on day 84 (P<0.05). The resistant group had higher mast cell counts in the abomasal mucosa than the susceptible group (P<0.05). These results indicate a protective TH2-mediated immune response against H. placei in the resistant group and a less protective TH1 response in the susceptible group.