Measuring the Genetic Diversity of Xanthomonas axonopodis pv. manihotis Within Different Fields in Colombia

ABSTRACT Cassava bacterial blight, caused by Xanthomonas axonopodis pv. manihotis, is a widespread disease that affects cassava (Manihot esculenta). We collected 238 X. axonopodis pv. manihotis strains by intensively sampling single fields in four edaphoclimatic zones (ECZs) in Colombia. DNA polymorphism of different X. axonopodis pv. manihotis populations was assessed by restriction fragment length polymorphism (RFLP) analyses, repetitive sequence-based polymerase chain reaction (rep-PCR), and amplified fragment length polymorphism (AFLP) assays. Genetic diversity, phenetic relationships among strains, and the coefficient of genetic differentiation were determined. All strains were tested for aggressiveness on the susceptible cassava cv. MCOL 1522. Strains were also tested for virulence on cassava differentials adapted to the strains' respective ECZs. Our study showed that the Colombian X. axonopodis pv. manihotis population has a high degree of genetic diversity. The hierarchical analysis of diversity showed genotypic differentiation at all levels, among ECZs, among fields within ECZs, and among strains within fields planted to several cassava genotypes. New RFLP haplotypes were detected, leading to the characterization of a new pathotype. Dendrograms from AFLP were more robust than those from RFLP data. A close association between the strains' geographical origin and DNA polymorphism was obtained using RFLP and AFLP data. We suggest that the host played a role in causing pathogen differentiation.     

Analysis of the association between the incidence of two spatially aggregated foliar diseases of strawberry

ABSTRACT Association of the incidence of leaf blight (caused by Phomopsis obscurans) and leaf spot of strawberry (caused by Mycosphaerella fragariae) was assessed at multiple scales in perennial plantings at several commercial farms over 3 years (1996 to 1998). For each field, the presence or absence of each disease was recorded from n = 15 leaflets in each of N approximately 70 evenly spaced sampling units, and the proportion of leaflets with blight, spot, and total disease (blight or spot) was determined. Individual diseases and total disease incidence were all well described by the beta-binomial distribution but not by the binomial distribution, indicating overdispersion of disease. The Jaccard similarity index was used to measure disease co-occurrence at the leaflet, sampling-unit, and field scales. Standard errors of this index for the lower two scales were obtained using the jackknife (resampling) procedure, and data randomizations were used to determine the expected Jaccard index for an independent arrangement of the two diseases, conditioned on the incidence and spatial heterogeneity of the observed disease data. Results based on these statistics showed that only 4 of 52 data sets at the leaflet level and no data sets at the sampling-unit level had Jaccard index values significantly different from that expected under an independent rearrangement of the two diseases. Rank correlation and cross-correlation statistics were calculated to determine the degree of covariation in incidence between the two diseases. Additionally, covariation between diseases was tested using a new procedure in the Spatial Analysis by Distance IndicEs (SADIE) class of tests. Covariation was detected in 21% of the data sets using rank correlation methods and in 15% of the data sets using the SADIE-based approach. The discrepancy between these two methods may be due to the rank correlation procedure not taking into account the effects of spatial pattern of disease incidence. There was no relationship between mean disease incidence per field of spot and blight or between degree of heterogeneity of the two diseases (as measured by theta of the beta-binomial distribution), demonstrating lack of covariation at the field scale. Incidence of leaflets with either disease (total disease incidence) could be well predicted using a linear combination of the estimated probabilities of leaf blight and leaf spot incidence based on independence of the two diseases. Heterogeneity of total disease incidence, measured with the estimated theta parameter of the beta-binomial distribution, could also be well predicted using a linear combination of the weighted theta values for leaf blight and leaf spot, with weights proportional to incidence of the individual diseases.     

Serological Methods for Detection of Polymyxa graminis, an Obligate Root Parasite and Vector of Plant Viruses

A purification procedure was developed to separate Polymyxa graminisresting spores from sorghum root materials. The spores were used as im-munogen to produce a polyclonal antiserum. In a direct antigen coating enzyme-linked immunosorbent assay (DAC ELISA), the antiserum could detect one sporosorus per well of the ELISA plate. In spiked root samples, the procedure detected one sporosorus per mg of dried sorghum roots. The majority of isolates of P. graminis from Europe, North America, and India reacted strongly with the antiserum. Interestingly, P. graminis isolates from the state of Rajasthan (northern India), from Pakistan, and an isolate from Senegal (West Africa) reacted weakly with the antiserum. The cross-reactivity of the serum with P. betae isolates from Belgium and Turkey was about 40% of that observed for the homologous isolate. There was no reaction with common fungi infecting roots or with the obligate parasite Olpidium brassicae. However, two isolates of Spongospora sub-terranea gave an absorbance similar to that observed with the homologous antigen. The DAC ELISA procedure was successfully used to detect various stages in the life cycle of P. graminis and to detect infection that occurred under natural and controlled environments. A simple procedure to conjugate antibodies to fluorescein 5-isothiocyanate (FITC) is described. Resting spores could be detected in root sections by using FITC-labeled antibodies. The potential for application of the two serological techniques for studying the epidemiology of peanut clump disease and for the characterization of Polymyxa isolates from various geographical origins is discussed.     

Forecasting Infections of the Leaf Curl Disease on Peaches Caused by Taphrina deformans

An Israeli model forecasting leaf curl disease on peaches caused by Taphrina deformans was validated in the Emilia-Romagna region of northern Italy, during a three-year period (1996–1998), in 13 cases (year × location × cultivar). When the peach trees are susceptible to infection, the model uses mathematical functions to calculate the risk of infection on the basis of weather conditions (daily rainfall greater than 10mm, and maximum air temperature greater than 5°C), and it forecasts periods of possible symptom appearance based on the length of incubation. Peach trees became susceptible to infection between the end of January and mid March, when the first leaf buds attained phenological stage C, i.e. appearance of leaf apex. The trees remained susceptible for at least 9 weeks: the last infection occurred in mid-May.Since most of the leaf curl onsets observed in the orchards fell within the range of model forecasts, the model proved to be accurate in signalling both the first seasonal infection and repeated infections during the primary inoculum season. Few errors occurred, caused either by conditions of rainfall and temperature lower than the thresholds fixed in the model, or by discrepancies between forecast and actual length of incubation. Infection occurred also at 3.1–3.5°C, and with 9.6mm rainfall. Thus, thresholds should not be accepted too rigorously, and perhaps temperature should not be considered as a limiting factor for infection under the conditions of the present work. The length of incubation showed high variability: it was 23 days long on average, with a 95% confidence interval ranging from 20 to 27 days, and extreme values of 9 and 33 days.     

MAGNETIC RESONANCE IMAGING CHARACTERISTICS OF THE FOOT IN HORSES WITH PALMAR FOOT PAIN AND CONTROL HORSES

Palmar foot pain is a common cause of lameness. Magnetic resonance imaging (MRI) has the potential to detect damage in all tissues of the equine foot, but an understanding of the differences in magnetic resonance (MR) images between feet from horses with and without palmar foot pain is required. This study aimed to describe MR findings in feet from horses with no history of foot-related lameness, and to compare these with MR findings in horses with lameness improved by palmar digital local analgesia. Thirty-four limbs from horses euthanized with a clinical diagnosis of navicular syndrome (ameness >2 months duration, positive response to palmar digital nerve blocks and absence of other forelimb problems) (Group L), and 25 feet from age-matched horses with no history of foot pain (Group N) were examined. For each anatomic structure, MR signal intensity and homogeneity, size, definition of margins, and relationships with other structures were described. Alterations in MR signal intensity and homogeneity were graded as mild, moderate, or severe and compared between Groups L and N. Results revealed that there were significant differences in MR images between Groups N and L. Multiple moderate-severe MR signal changes were present in 91% of limbs from Group L and moderate (none were graded severe) in 27% of limbs from Group N. In most Group L limbs, more than three structures and frequently six to eight structures were abnormal. Concomitant abnormalities involved most frequently the deep digital flexor tendon, distal sesamoidean impar ligament, navicular bone, collateral sesamoidean ligament, and navicular bursa (with significant associations in severity grade between these structures), sometimes with involvement of the distal interphalangeal joint and/or its collateral ligaments. It was concluded that findings on MR images were different between horses with and without foot pain, and that pain localized to the foot was associated with MR changes in a variety of structures, indicating that damage to several structures may occur concurrently and that MR imaging was useful for evaluation of foot pain.     

Endometrial Adenocarcinoma and Mucometra in a 6-year-old Alaska Malamute Dog

A 6-year-old female Alaska Malamute dog was presented for evaluation of abdominal enlargement referred by a local veterinarian. On the history, the owner complained of chronic abdominal enlargement initiated more than 4 months ago, reduced appetite, occasional vomiting and general dullness. He also complained of greenish mucous intermittent vaginal discharge starting 10 days ago. The bitch was chronically treated with medroxiprogesterone acetate. A laparatomy was performed and fluid in the abdomen was found and aspirated during the surgery. Also a very fluid-filled distended uterus and a mass in the distal part of the left uterine horn were found. The mass was encapsulated by the omentum, but areas of necrosis and calcification were identified. Histopathological diagnosis was endometrial adenocarcinoma.     

Panniculitis in a horse with peripancreatitis and pancreatic fibrosis.

A 22-year-old pinto mixed breed mare was admitted for evaluation of severe colic signs and gastric reflux. Multiple nonpainful, variably sized hard masses were palpated in the subcutis over the thorax, abdomen, and hindquarters of the horse. The mare was diagnosed with sterile peritonitis and had systemically high gamma-glutamyltransferase, amylase, and lipase. Three days into treatment she became febrile with signs of persistent and mild abdominal discomfort; euthanasia was elected. Necropsy revealed peripancreatitis, pancreatic fibrosis, abdominal steatitis and panniculitis. Panniculitis associated with peripancreatitis has been described in humans and dogs but not in horses. Pathogenic models for this entity are discussed.     

Test strategies in bovine viral diarrhea virus control and eradication campaigns in Europe.

Several European countries have initiated national and regional control-and-eradication campaigns for bovine viral diarrhea virus (BVDV). Most of these campaigns do not involve the use of vaccines; in Germany, vaccination is used only in states in which it is considered necessary because of high BVDV prevalence. In European countries without organized BVDV control programs, vaccination is commonly used to control BVDV. Diagnostic test strategies are fundamental to all control-and-eradication campaigns; therefore, the purpose of this review is to describe how the available diagnostic tests are combined into test strategies in the various phases of control-and-eradication campaigns in Europe. Laboratory techniques are available for BVDV diagnosis at the individual animal level and at the herd level. These are strategically used to achieve 3 main objectives: 1) initial tests to classify herd status, 2) follow-up tests to identify individual BVDV-infected animals in infected herds, and 3) continued monitoring to confirm BVDV-free status. For each objective or phase, the validity of the diagnostic tests depends on the mode of BVDV introduction and duration of infection in test-positive herds, and on how long noninfected herds have been clear of BVDV. Therefore, the various herd-level diagnostic tools--such as antibody detection in bulk milk or in blood samples from young stock animals, or BVDV detection in bulk milk--need to be combined appropriately to obtain effective strategies at low cost. If the individual diagnostic tests are used with due consideration of the objectives of a specific phase of a BVDV control program, they are effective tools for controlling and eradicating BVDV in regions not using vaccination and where vaccination is a part of the control or eradication program.     

Glomangioma in the Prepuce of a Dog

A case of preputial glomangioma in an 8-year-old Springer Spaniel dog is reported. The dog presented a solitary nodular mass, approximately 3.5 x 5.2 cm in diameter, located on the lateral wall of the prepuce at the base of the penis. Histologically, the excised tumour mass consisted of proliferating round epithelioid cells arranged in a cordon-like structure around small blood vessels. The neoplastic cells were positive for alpha smooth muscle actin and negative for cytokeratin, desmin, S-100 protein and neurone-specific enolase. Based on these pathologic features, the tumour was diagnosed as glomangioma, a neoplasm extremely rare in dogs. To our knowledge, this is the first report of preputial glomangioma in the dog.     

Post-mortem Examination of Sows Genital Organs Culled for Reproductive Disturbances and Immunohistochemical Studies on ERα and PR A Receptors in the Anoestral Sows Uterus

The aim of present study was post-mortem examination of ovaries, uterus and plasma oestradiol-17beta (E2) and progesterone (P4) concentrations in the blood of sows with reproductive disturbances and the distribution of oestradiol receptor (ERalpha), as well as progesterone (PR-A) in the anoestrous sows uteri. Reproductive organs of 150 crossbred sows (Lithuanian White x Danish Landrace) culled for the reasons of reproductive disturbances, were collected in local abattoir over a period of 3 months (September-November). Organs were assessed to determine the stage of the oestrous cycle or anoestrus and their development. Blood samples were collected for E2 and P4 analysis from the jugular vein 1 h prior to slaughter. For this study uterine samples only from pathological anoestrous sows were subjected to immunohistochemical staining to assess the distribution of ERalpha and PR-A in surface epithelium, subepithelial connective tissue, glandular epithelium and myometrium. Macroscopic examination of the ovaries showed that 68.7% sows had active cycling ovaries, 26.6% sows were anoestrus, ovaries were small without CL, and 4.7% of sows had multiple follicular cysts. In anoestrous sows (n = 27) the number and intensity of the nuclear staining of ERalpha varied between different uterine tissue compartments. The highest number (>80%) and the strongest intensity (+++) of positively stained cells for ERalpha was seen in myometrium and glandular epithelium. In other uterine wall compartments the number and intensity of positively stained for ERalpha nuclei was lower (+/++). The PR-A was absent from all tissue compartments. The intensity of the nuclear staining for ERalpha varied not only between the different uterine compartments but also between the sows. The 11.1% of the sows presented ERalpha in surface epithelium, 74.1% of the sows in glandular epithelium and 63.0% of sows in the myometrium.