Salivary Cortisol, Equine Characteristics, and Management Factors Associated With Strongyle-Type Egg Shedding of Ohio Horses.

Identification of factors associated with parasite infestation in horses could reduce frequency of anthelmintic administration and slow development of anthelmintic resistance. The study aim was to evaluate management factors, equine characteristics, and equine salivary cortisol concentrations for association with strongyle-type egg shedding levels. As immunocompromised horses appear to be more susceptible to parasite infestation, it was hypothesized that salivary cortisol concentration could be associated with parasite egg shedding. Saliva and fecal samples were collected from 200 horses across the state of Ohio. Equine management questionnaires were administered to barn managers. Fecal egg counts were performed with a modified Stoll method, and saliva samples were analyzed for salivary cortisol with an enzyme-linked immunosorbent assay (ELISA). A total of 24 variables were tested for association with fecal egg count results (dichotomized with 50 eggs per gram as a cutoff) using chi-square test of independence, Fisher's exact test, independent t-test, or Mann Whitney U test. The sample of horses was partitioned into tertiles based on salivary cortisol concentrations (i.e., low <0.052 µg/dL, medium 0.052-0.0712 µg/dL, and high >0.0712 µg/dL). Variables with P < .30 were analyzed for association with fecal egg count with multiple logistic regression. The final logistic regression model (P< .05) included horse age, horse salivary cortisol concentration, and pasture mowing frequency. Older horses had higher adjusted odds of having <50 EPG. Pasture mowing frequency of at least once per week increased the adjusted odds of <50 EPG. Horses with cortisol concentrations in the highest tertile had increased adjusted odds of having <50 EPG.

Ips pini (Curculionidae: Scolytinae) is a vector of the fungal pathogen, Sphaeropsis sapinea (Coelomycetes), to Austrian pines, Pinus nigra (Pinaceae).

Sphaeropsis sapinea (Fr.:Fr.) Dyko and Sutton, is among the most common and widely distributed pathogens of conifers worldwide. S. sapinea is disseminated over short distances by rain splash and moist wind, but significant knowledge gaps regarding long-range dispersal remain. Our objective was to determine whether or not the pine engraver beetle, Ips pini Say, is a vector of the pathogen onto Austrian pines (Pinus nigra Arnold). In 2004 and 2005, individuals of I. pini were collected with pheromone traps at two locations in central Ohio (197 and 1,017 individuals for 2004 and 2005, respectively) and screened for the presence of S. sapinea. In the field, fresh logs of Austrian pine were baited with pheromone lures, mechanically wounded, or left undisturbed. After 2 mo, logs were evaluated for insect feeding and the presence of S. sapinea along beetle galleries. Fresh logs were also inoculated in the greenhouse with adult I. pini that were either artificially infested or uninfested with S. sapinea spores to determine vectoring potential. Phoresy rates for individual collections ranged from 0 to 4.1%; average rates were 1.5 and 2.0% for 2004 and 2005, respectively. Isolation frequencies of S. sapinea from baited (15 +/- 5%) and unbaited logs (3 +/- 1%) differed significantly (P=0.009). I. pini was also capable of transmitting the pathogen under controlled conditions. Based on phoresy rates, association, and artificial inoculation studies, we conclude that I. pini is able to transmit S. sapinea to Austrian pine stems.

Heritability of Latent Period Estimated from Wild-Type and Selected Populations of Puccinia triticina.

ABSTRACT Durability of partially resistant wheat cultivars to wheat leaf rust depends on the amount of genetic variation in parasitic fitness within populations of the pathogen Puccinia triticina. To assess the durability of partial resistance, selection experiments were used to explore quantitative variation in parasitic fitness of P. triticina. Fungal populations 881-WT and 882-WT were selected for shortened latent period on partially resistant cvs. CI 13277 and Sw 72469-6 for multiple generations. Fitness components were measured for wild-type and selected fungal populations. Responses to selection and selection differentials were calculated, and broad-sense, realized heritabilities for latent period were estimated for wild-type fungal populations on CI 13227 and on Sw 72469-6. Selected populations had fitness characteristics, not limited to latent period, that could provide greater fitness in nature. Generally, more cycles of selection had greater effects on fitness. In particular cases, selected populations on a partially resistant cultivar had values for latent period, uredinium area, and sporulation no different from those of a susceptible host-pathogen combination. Heritabilities of latent period of populations 881-WT and 882-WT on CI 13227 or populations 881-WT and 882-WT on Sw 72469-6 ranged from 0.65 to 0.76 and 0.17 to 0.24, respectively. Our results suggest the variation to overcome quantitative host resistance exists in extant populations of P. triticina. In addition, because more of the variation in latent period for populations of P. triticina on CI 13227 was genetic than for populations on Sw 72469-6, CI 13227 is likely to be more vulnerable to pathogen adaptation despite its exceptionally long latent period.

Host Resistance to Monilinia vaccinii-corymbosi in Flowers and Fruits of Highbush Blueberry.

Monilinia vaccinii-corymbosi, the causal agent of mummy berry disease, infects blueberry flowers via the gynoecial pathway. To describe the expression of host resistance in highbush blueberry (Vaccinium corymbosum), fungal growth in the styles and colonization of the locules were compared among five blueberry cultivars in a series of controlled greenhouse experiments. Styles were harvested 1 and 4 days postinoculation, and the length colonized by hyphae was determined using fluorescence microscopy. At 8 weeks after inoculation, fruit were harvested and scored for the presence of hyphae in the locules. The infection frequency of styles ranged from 0.33 to 0.71, and only cv. Weymouth had significantly lower infection frequency than the other cultivars. The mean length of the colonized portion of the stylar canal ranged from 0.126 to 0.434 mm after 1 day and 1.62 to 3.59 mm after 4 days. Hyphae in the styles of cv. Weymouth exhibited the least growth, whereas hyphae in the styles of cultivars Jersey and Rancocas were significantly longer. The distance of style penetrated for cultivars Bluecrop and Coville was intermediate. The mean disease incidence of locules differed significantly. Values for cultivars Weymouth and Jersey were the smallest (0.038 and 0.039) and largest (0.249 and 0.236), respectively. The results demonstrate that a component of resistance to infection by M. vaccinii-corymbosi is expressed during growth in the gynoecial pathway.

Relationship Among Genes Conferring Partial Resistance to Leaf Rust (Puccinia triticina) in Wheat Lines CI 13227 and L-574-1.

ABSTRACT This study describes the segregation of genes for resistance to the fungus Puccinia triticina in a cross between partially resistant wheat lines L-574-1 and CI 13227 with two and four genes for resistance, respectively. The objectives of this study were to use parental, F(1), F(2), and backcross populations to quantify maternal effects, degree of dominance, and transgressive segregation, and to determine whether CI 13227 and L-574-1 share any resistance genes for long latent period or small uredinia. In two experiments conducted in the greenhouse, the uppermost leaf of adult wheat plants was inoculated prior to heading with P. triticina. On days 6 to 21 after inoculation, the number of uredinia that erupted from the leaf surface was counted and used to calculate the mean latent period (MLP). The length and width of five arbitrarily selected uredinia were measured and used to calculate uredinium area. Midparent values, degree of dominance, and broad-sense heritability were calculated for MLP and uredinium area. For experiment A, MLP values for CI 13227, L-574-1, F(1), and F(2) generations were 12.2, 10.5, 10.2, and 10.6 days, respectively. For experiment B, MLP values for CI 13227, L-574-1, F(1), F(2), backcross to CI 13227, and backcross to L-574-1 were 12.3, 10.0, 10.6, 10.8, 11.1, and 10.0 days, respectively. The inheritance of long latent period was partially recessive, and no maternal effect was present (P = 0.62 to 0.87 for the comparison of means in reciprocal crosses). Broad-sense heritability for MLP ranged from 0.72 to 0.74, and there was transgressive segregation in the F(2) and backcross populations. Uredinia of the F(1) generation were slightly larger than uredinia for CI 13227. The inheritance of uredinium size was partially dominant, and no maternal effect was present (P = 0.5 to 0.63). Broad-sense heritability for uredinium area ranged from 0.36 to 0.73 and transgressive segregation was present in the F(2) and backcross populations. The results for MLP indicate that lines CI 13227 and L-574 likely share one gene for resistance (based on F(1) values) but not two genes (based on the presence of transgressive segregation). CI 13227 and L 574-1 appear to have at least one gene difference for uredinium area. The linear relationship between uredinium area regressed onto MLP was significant (P < 0.001) and r(2) values ranged from 0.14 to 0.26. These results indicate that the resistance in CI 13227 and L-574-1 could be combined to create wheat cultivars with greater partial resistance than that possessed by either parent based on MLP or uredinium size.

First Report of Phytophthora insolita and P. inflata on Rhododendron in Ohio.

During August 2003, we conducted a statewide survey of rhododendrons to determine if Phytophthora ramorum was present in Ohio ornamental nurseries. In total, 240 samples were randomly collected in 12 nurseries throughout Ohio from rhododendrons showing foliar necrotic lesions and twig dieback symptoms. The samples yielded 51 Phytophthora spp. isolates on PARP-V8 agar. The internal transcribed spacer (ITS) region of all isolates was amplified using the universal primers ITS1 and ITS4 and was sequenced. Consensus sequences from sense and antisense were then blasted against the GenBank database, allowing for the identification to species of ˜80% of all isolates. These identifications, and the ˜20% unknowns, were confirmed using blind morphological tests on the basis of the following parameters: colony morphology; shape and dimensions of sporangia and type of papillae; dimensions of oogonia and oospores; type and position of antheridia; presence or absence of chlamydospores; presence or absence and morphology of hyphal swellings; and growth rate at 35°C according to the Revisited Tabular Key of the species of Phytophthora (1). No P. ramorum was detected among the isolates; however, P. cactorum, P. citricola, P. citrophthora, and P. nicotianae were detected. We also found two occurrences of P. inflata Caros & Tucker and one of P. insolita Ann & Ko. (P. inflata: e-value ≤e-179, identities ≥95%; P. insolita: e-value = 0.0; identities = 95%.) P. inflata was isolated from two tissue types, a dead twig and a necrotic leaf tip. P. insolita was isolated from a necrotic leaf tip. Identity of the two species was confirmed morphologically using the parameters listed above as well as the following measurements (N = 40; all in µm) (1): P. inflata - sporangia: 40 × 24 ([24 to 68] × [18 to 34]); oogonia: 34.6 (28 to 40); oospores: 30.8 (25 to 38); P. insolita - sporangia: 42 × 28 ([34 to 56] × [22 to 38]); oogonia: 32 (26 to 36); oospores: 26 (22 to 30). Koch's postulates were satisfied by inoculating two rhododendron plants (cvs. PJM and Nova Zembla) with the putative pathogens. On each plant, each of three leaves was pierced with a dissecting needle and was inoculated by placing a 0.5-cm-diameter plug of mycelium that was taken from the margin of a colony actively growing on PARP-V8 agar on the wound. The inoculum was retained using clear adhesive tape. A similar procedure was used for twigs. Controls consisted of inoculations with sterile PARP-V8 agar medium. Both cultures of P. inflata and P. insolita produced necrotic lesions in all inoculations on both tissue types within 1 week, and they were reisolated from the margins of lesions on PARP-V8. The lesion margin was at least 2 cm away from the inoculum plug in leaf inoculations and several centimeters in twig inoculations. To our knowledge, this is the first report of P. inflata and P. insolita occurring on rhododendron and the first time P. insolita has been reported outside of Southeast Asia where it has been recovered only from soil. Reference: (1) D. J. Stamps et al. Mycol. Pap. No. 162. CAB Int. Mycol. Inst. Wallingford, UK, 1990.