Amplified fragment length polymorphism as a tool for molecular characterization of almond germplasm: genetic diversity among cultivated genotypes and related wild species of almond,and its relationships with agronomic traits

Amplified fragment length polymorphism (AFLP) analysis is a rapid and efficient method for producing DNA fingerprints and molecular characterization. Our objectives were to: estimate genetic similarities (GS), marker indices, and polymorphic information contents (PICs) for AFLP markers in almond cultivars; assess the genetic diversity of almond cultivars and wild species, using GS estimated from AFLP fingerprints and molecular characterization; and facilitate the use of markers in inter-specific introgression and cultivar improvement. The genetic diversity of 45 almond cultivars from Iran, Europe, and America, were studied assaying 19 primer combinations. In addition, several agronomic traits were evaluated, including flowering and maturity times, self-incompatibility, and kernel and fruit properties. Out of the 813 polymerase chain reaction fragments that were scored, 781 (96.23%) were polymorphic. GS ranged from 0.5 to 0.96, marker indices ranged from 51.37 to 78.79, and PICs ranged from 0.56 to 0.86. Results allowed the unique molecular identification of all assayed genotypes. However, the correlation between genetic similarity clustering as based on AFLP and clustering for agronomic traits was low. Cluster analysis based on AFLP data clearly differentiated the genotypes and wild species according to their origin and pedigree, whereas, cluster analysis based on agronomic data differentiated according the pomological characterization. Our results showed the great genetic diversity of the almond cultivars and their interest for almond breeding.     

Spatial structure of genetic variation within populations of forest trees

The spatial pattern and structure of genetic variation are important aspects of the population genetics of forest stands. Combined with limits to seed and pollen dispersal, spatial structure affects the level of inbreeding and the action of natural selection. The genetic constitution of stand regeneration, following different forestry practices, is also influenced by spatial structure. For example, natural regeneration with seed trees involves sampling seed trees from a stand that may be genetically nonhomogeneous. This paper reviews theoretical and empirical results on spatial patterns of genetic variation, produced under limited gene flow and selection, in terms of recently developed spatial statistics (e.g., spatial autocorrelation). Genetic correlations in samples from spatially structured populations are also described, as well as how spatial samples can be used to characterize the structure of genetic variation, and how inferences can be made about (spatially distributed) components of fitness and yield.     

Assessing the effect of a single dose florfenicol treatment in feedlot cattle on the antimicrobial resistance patterns in faecal Escherichia coli

The objective of this clinical trial was to examine the effect of a single dose of florfenicol on antimicrobial resistance patterns in faecal E. coli of feedlot steers. Steers (n = 370), were purchased from two sources and housed in outdoor concrete floored pens. Two cattle from each pen (n = 42 pens, 84 cattle) were randomly selected for faecal sampling at study day 1, 14, 28, and 42. One sampled animal from each of 21 pens was randomly selected to receive a single 39.6 mg/kg dose of florfenicol subcutaneously at study day 11. Ten lactose positive colonies were isolated from faecal swabs and tested for antimicrobial resistance to 11 antimicrobials using the disk diffusion method. Zones of inhibition were grouped using cluster analysis and clusters were ordered by increasing multiple resistance. A cumulative logistic regression model using generalized estimating equations was used to assess factors associated with increasing levels of multiple resistance. Immediately post-treatment, all isolates obtained from treated cattle belonged to multiple resistant clusters containing chloramphenicol resistance. Though less pronounced in later sampling, resistance to chloramphenicol and other antimicrobials persisted. Antimicrobial treatment, sampling time and animal source, as well as interactions between these variables, were important predictors of the odds of E. coli belonging to a more resistant cluster. A very clear but transitory shift to increasingly multiple resistant faecal E. coli in response to florfenicol treatment was observed. There was no indication of horizontal transfer of resistant E. coli between steers. Level of resistance was influenced by complex interaction of animal source and previous management.     

Medication‐related osteonecrosis of the jaw after long‐term bisphosphonate treatment in a cat

A 12‐year‐old, neutered female, domestic medium hair cat was evaluated for a nonhealing, oral mucosal ulceration. The cat had a history of idiopathic hypercalcemia that had been treated with a bisphosphonate for 41 months. Oral examination identified exposed maxillary bone adjacent to a previous extraction site. Histopathology of the exposed bone and associated mucosa was most consistent with medication‐related osteonecrosis of the jaw. Treatment involved both medical and surgical interventions. Oral mucosal healing occurred after 6 months of treatment.     

Evaluation of the prevalence and onset of lung lesions and their impact on growth of lambs

OBJECTIVE: To determine the prevalence and temporal onset of lung lesions in lambs and the impact of lung lesions on growth of affected lambs. ANIMALS: 259 crossbred wether lambs from a single flock in the upper Midwestern United States. PROCEDURE: An observational study was conducted. Lambs born in the spring and fall were slaughtered at finished weight or at a predetermined time point. Lungs of each lamb were examined and classified as normal, moderate lesions (consolidation > 5% but < or = 50% of any lobe), or severe lesions (consolidation > 50% of any lobe). Data were examined to detect effects of prevalence or severity of lung lesions on growth and carcass traits. RESULTS: 57 of 89 (64%) spring-born lambs had lung lesions characterized by consolidation of lung tissue. A small number of lambs had pulmonary adhesions or active abscesses. In contrast, only 31 of 108 (29%) fall-born lambs had lung lesions. Severe lung lesions were associated with a significant reduction in average daily gain. Severe lung lesions were not detected until the middle of the finishing period and were associated with culture of Mannheimia haemolytica or Pasteurella multocida. CONCLUSIONS AND CLINICAL RELEVANCE: Analysis of results indicates that the prevalence of severe lung lesions can be quite high in lambs. Severe lung lesions can lead to greatly decreased growth performance of lambs.     

Comparison of two DNA extractions and nested PCR, real-time PCR, a new commercial PCR assay, and bacterial culture for detection of Mycobacterium avium subsp. paratuberculosis in bovine feces.

In this study, 5 combinations of 2 DNA extractions and 3 polymerase chain reaction (PCR) techniques were compared with culture for the detection of Mycobacterium paratuberculosis directly from bovine feces. These combinations included a new commercial extraction technique combined with a commercial PCR/Southern blot technique, nested PCR (nPCR), or real-time PCR, and a university-developed extraction combined with nPCR or real-time PCR. Four of the 5 combinations had statistically similar sensitivities between 93% and 100% and specificity between 95% and 100%, when compared with culture results from 63 bovine fecal samples. These results indicated that using a commercial extraction with a commercial PCR/Southern blot, nPCR, or real-time PCR, or a university-developed extraction with real-time PCR would result in similar sensitivities to culture for the identification of M. paratuberculosis from bovine feces and are valid alternatives to culture.     

Impact of trace mineral source on beef replacement heifer growth,reproductive development,and biomarkers of maternal recognition of pregnancy and embryo survival

Trace minerals are known to play important roles in early embryo development. The study objective was to determine effects of trace mineral source on heifer reproductive performance. Beef heifers (n = 129) were randomly assigned to one of two treatments. From weaning through breeding, all heifers were individually fed a basal diet supplemented with cobalt (Co), copper (Cu), manganese (Mn), and zinc (Zn) either from organic sources (COMP; Cu, Mn, and Zn amino acid complexes and Co glucoheptonate; Availa-4, Zinpro Corporation, Eden Prairie, MN) or inorganic sources (INORG; Cu, Mn, and Zn hydroxychlorides; Intellibond C, M, and Z, Micronutrients, Indianapolis, IN) and Co as CoSO₄. Blood samples and a reproductive tract score (RTS) were collected to determine pubertal status. All animals were synchronized and artificially inseminated. Pregnancy status was determined by lymphocyte gene expression, circulating concentrations of pregnancy-associated glycoproteins (PAGs), and by transrectal ultrasonography after artificial insemination. Embryonic loss was defined as when a previously pregnant animal was subsequently diagnosed not pregnant. Data were analyzed using the MIXED procedure in SAS. Puberty (P = 0.44), pelvic area (P = 0.74), RTS (P = 0.49), and estrus expression (P = 0.82) were not influenced by treatment. There was no effect of treatment (P = 0.37) or treatment by time (P = 0.19) on pregnancy, but there was a tendency (P = 0.13) for decreased embryonic loss among COMP heifers (27 ± 6%) compared to INORG heifers (38 ± 6%). There was a treatment by pregnancy status by time interaction (P < 0.01) on circulating PAG concentrations with PAG concentrations tending (P = 0.08) to be greater on day 25 among heifers in the COMP treatment compared to heifers in the INORG group. In summary, source of trace mineral did not affect puberty, RTS, pelvic area, or overall pregnancy success, but feeding complexed trace minerals tended to increase circulating PAG concentrations and embryo survival.     

A single pasture limited treatment approach to estimate production loss from gastrointestinal nematodes in grazing stocker cattle

The objective of this study was to evaluate the use of a single pasture, limited treatment methodology to assess the impact of gastrointestinal nematodes on weight gain in grazing cattle. From a group of 450 British crossbred, yearling spayed heifers, 60 animals were randomly selected (prospective randomization) prior to placement on summer pasture. Each of these 60 animals was weighed, a fecal sample obtained for nematode egg count and a uniquely numbered ear tag applied. A randomly assigned group of 30 received treatment with one ivermectin sustained release bolus, while the remaining 30 served as non-treated controls. The treatment and control groups rejoined the remaining non-treated 390 animals, and were grazed as a single group for 165 days. At grazing conclusion, treatment and control cattle were individually weighed, and fecal samples obtained for nematode egg counts. Treatment was associated with a 0.064 kg per grazing day gain increase, or a 10 kg increase over the grazing season (P = 0.02). Nematode egg counts at grazing initiation were not different between treatment and control (P = 0.30), though egg counts in treated cattle at study end were lower than control (P < 0.0001). Results of this study support the use of a single pasture limited treatment approach for measuring the effect that internal nematodes have on weight gain in grazing cattle under commercial range conditions.     

Survey and molecular detection of phytoplasmas associated with potato in Romania and southern Russia

In recent years, emerging phytoplasma diseases of potato (Solanum tuberosum L.) have increasingly become important in central and eastern Europe. Accurate identification of phytoplasmas and their insect vectors is essential to developing effective management strategies for diseases caused by these plant pathogens. Potato phytoplasma diseases in Europe were for a long time diagnosed only on the basis of visual symptoms. However, this approach is not very reliable and the use of modern molecular techniques such as polymerase chain reaction (PCR) is required in order to accurately determine the etiology of these phytoplasma diseases. A survey and identification of phytoplasmas associated with potato crops in Romania and southern Russia were conducted based on modern molecular techniques. Symptomatic potato plants were collected from several fields and tested for phytoplasmas by PCR. Also, selected crops and weeds in the vicinity of these potato fields were sampled and tested for phytoplasmas. Stolbur (“Candidatus Phytoplasma solani”; 16SrXII-A) was the only phytoplasma detected in potato and adjacent crops, including tomato (Solanum lycopersicum), pepper (Capsicum annuum), eggplant (Solanum melongena), and beet (Beta vulgaris). This phytoplasma was also detected in weeds, particularly Convolvulus arvensis, Cuscuta sp., and Euphorbia falcata. Genotyping of obtained stolbur isolates on tuf genes revealed that they all had the same RFLP profile corresponding to the tuf-type ‘b’ (VK Type II). Stolbur-affected potato plants produced a large number of spongy tubers that resulted in commercially unacceptable potato chips upon processing.     

Important genetic diversity of ‘<Emphasis Type="Italic">Candidatus</Emphasis> Phytoplasma solani’ related strains associated with bois noir grapevine yellows and planthoppers in Azerbaijan

Bois noir (BN) is an important grapevine yellows endemic to the Euro-Mediterranean basin caused by ‘Candidatus Phytoplasma solani’ (‘Ca. P. solani’), a non culturable plant pathogenic Mollicute. Bois noir symptoms could be associated with ‘Ca. P. solani’ in two Azerbaijanian vineyards where disease incidence and severity were recorded for five local Vitis vinifera cultivars. In order to gain insight into the epidemiology of Bois noir in Azerbaijan, ‘Ca. P. solani’ isolates infecting plants were characterized by multi-locus sequence analysis and their secY and stamp gene sequences compared to that of the strains detected in other plants and in local Cixiidae planthoppers. Genotypes were determined for two non-ribosomal house-keeping genes, namely tuf and secY, as well as two variable markers namely Stamp and mleP1 genes, that respectively encode the antigenic membrane protein AMP and a 2-Hydroxycarboxylate transporter. The Azerbaijanian BN phytoplasma isolates corresponded to three tufB and secY genotypes. A finer differentiation of Azerbaijanian ‘Ca. P. solani’ isolates was obtained with mleP1 as five different mleP1 genetic variants were found. Finally, Stamp gene allowed differentiating four new genotypes in grapevine among the 10 new Stamp genotypes detected in various plants in Azerbaijan. The preliminary survey for infected insects conducted in northern Azerbaijan, led to the identification of Hyalesthes obsoletus and Reptalus noahi as potential vectors for two ‘Ca. P. solani’ new genotypes phylogenetically distant from the known genetic clusters. Altogether these results indicate an important genetic diversity of BN phytoplasmas in Azerbaijan that certainly result from spread through local insect vectors.