A duplex real-time polymerase chain reaction assay for the detection of and differentiation between Dirofilaria immitis and Dirofilaria repens in dogs and mosquitoes

This study describes a duplex real-time polymerase chain reaction (PCR) assay for the detection and differentiation between Dirofilaria immitis and Dirofilaria repens in dog blood and mosquitoes. Regions of a cytochrome oxidase 1 (cox1) mitochondrial DNA fragment and the second internal transcribed spacer (ITS-2) of nuclear ribosomal DNA were amplified from microfilariae and adult worm samples, using a sensitive SsoFast? EvaGreen(?) based real-time PCR method coupled with melting-curve analysis. The limit of the real-time PCR in detecting microfilaria and adult worm DNA was also tested both in dog blood and in artificially infected microfilarial. Two peaks at different melting temperatures (T(m)) for D. immitis (mean ± SD=75.7 ± 0.3°C) and D. repens (mean ± SD=70 ± 0.7°C), respectively, were obtained for microfilarial and adult positive controls of both species when examined separately and together. The real-time PCR protocol was also efficient in detecting microfilarial and adult DNA of both species when tested in samples spiked with DNA from Aedes albopictus, in Aedes aegypti experimentally infected by D. repens and in Culex pipiens naturally infected by D. repens and D. immitis. The high sensitivity of real-time PCR confirmed its reliability in detecting small amounts of genomic DNA either in dog blood or mosquitoes (2.5 pg/μl and 3 × 10(-1)pg/μl for D. immitis and D. repens, respectively). This assay is proposed as a tool for the epidemiological surveillance of the two most important Dirofilaria species in areas where they are endemic and sympatric.     

Application of 10% imidacloprid/50% permethrin to prevent Ehrlichia canis exposure in dogs under natural conditions

Canine monocytic ehrlichiosis (CME) caused by Ehrlichia canis is the most known canine tick-borne disease (TBD) spread throughout the world. Preventing tick bites is a priority to reduce the risk of TBDs and it was the aim of the present study to evaluate the efficacy of a combination of imidacloprid 10% and permethrin 50% (ImPer) (Advantix; Bayer AG, Germany) in a spot-on formulation to control CME under field conditions. On January-March 2005, 845 dogs from two kennels in southern Italy (kennels of Bari (KB)- and Ginosa (KG)), with a history of tick infestation were initially tested by serology and PCR assay for E. canis infection. Data on Leishmania infantum infection were also available from a previous study carried out on the same dog population. One hundred twenty-six dogs (14.9%) presented anti-E. canis antibodies with a relative prevalence of 15.6% (n=65 dogs in KB) and 14.2% (n=61 dogs in KG). Five hundred thirty-five animals found negative both for E. canis and L. infantum infections were enrolled in three groups (Group A--treated with ImPer once a month; Group B--treated every 2 weeks; and Group C--untreated control animals) and monitored for E. canis infection by serology and PCR in November 2005 (first follow-up) and in March 2006 (second follow-up). The E. canis infection was serologically revealed, at the first and/or second follow-up, in 26 animals from Group C in KB and KG (mean incidence density rate (IDR), 13.24%) while in none of the animals from Group A (KB and KG) and only in one animal from Group B (IDR 1.13%) in KG. The final protection efficacy of ImPer ranged from 95.57% to 100% in Groups B and A. At PCR only 15 dogs from KG were positive for Rickettsiales only at the first follow-up and at the sequence analysis two (both in Group C) revealed 100% homology with E. canis sequences while 13 with Anaplasma platys. Four out of 13 A. platys PCR-positive dogs were also seropositive for E. canis at one or both follow-ups. ImPer, by virtue of its repellent and acaricidal activity against ticks, has been shown to be efficacious to prevent E. canis infection in treated dogs living under natural conditions in endemic areas.     

3 TESLA MAGNETIC RESONANCE IMAGING OF THE OCCIPITOATLANTOAXIAL REGION IN THE NORMAL HORSE

The aim of this study was to describe the appearance of the ligamentous structures of the occipitoatlantoaxial (OAA) region in the normal horse by 3 tesla (3T) magnetic resonance imaging (MRI). The MRI images of the longitudinal odontoid ligament, tectorial membrane, dorsal and ventral atlantoaxial ligaments, dorsal atlantooccipital membrane with its reinforcing ligaments, and the lateral atlantooccipital ligaments of 10 horse cadavers were evaluated. All ligaments and membranes were identified in all planes, except for the lateral atlantooccipital ligament in the sagittal plane due to its cranioventrolateral course. All were iso to mildly hypointense to musculature of the neck in T1W with the exception of the tectorial membrane that was moderately hypointense; moderately hypointense in PD‐SPIR, and markedly hypointense (isointense to cortical bone) in T2W. The PD‐SPIR was the best sequence to identify all ligaments and membranes from their cranial and caudal attachments. The longitudinal odontoid ligament, ventral atlantoaxial ligament, and reinforcing bands of the dorsal atlantooccipital membrane presented a characteristic striped heterogeneous signal behavior thought to be due to fibrocartilaginous content. The remaining ligaments and membranes showed homogeneous signal intensity. Special anatomical features in this species such as the fan‐shaped longitudinal odontoid ligament, absence of the transverse ligament and presence of the ventral atlantoaxial ligament were documented. Ligamentous structures that stabilize the equine OAA region were described with MRI in this study and these findings could serve as an anatomic reference for those cases where instability of this region is suspected.     

Characterization of the LH peak after short and long fixed‐time artificial insemination protocols in sheep raised in the tropics

The aim of this study was to evaluate the peak in luteinizing hormone (LH) and the pregnancy rate of sheep (Texel × Santa Inês) in the tropics using short‐ (6 days) and long‐term (12 days) progesterone protocols followed by artificial insemination (AI) both in and out of the breeding season. Experiment 1 was conducted within (IN) the breeding season (autumn, n = 36), and experiment 2 was conducted outside (OUT) of the breeding season (spring, n = 43). In each experiment, the sheep were divided into two groups (6 or 12 days) according to the duration of treatment with a single‐use progesterone release vaginal device (CIDR^®, Pfizer, São Paulo, SP, Brazil), and blood samples were collected from 10 animals per group every 4 hr to measure the LH and progesterone concentrations. In the spring, the characteristics of the LH peak did not differ between groups; but in the autumn, there were differences between groups at the beginning (G‐6 IN: 36.44 ± 5.46 hr; G‐12 IN: 26.57 ± 4.99 hr) and end of the LH peak (G‐6 IN: 46.22 ± 7.51 hr; G‐12 IN: 34.86 ± 8.86 hr). The results showed alterations in the LH peak during the breeding season only in the sheep undergoing the short‐term protocol.     

Cytologic examination of fine‐needle aspirates from mammary gland tumors in the dog: diagnostic accuracy with comparison to histopathology and association with postoperative outcome

Background: Mammary tumors are the most common neoplasms in female dogs. Malignant tumors may carry a poor prognosis and necessitate surgery. Few data are available on the value of cytologic examination as a diagnostic or prognostic tool for mammary tumors in dogs. Objectives: The objectives of this study were to determine whether cytologic findings in fine‐needle aspirate specimens of canine mammary tumors correlate with histopathologic results and whether the cytologic diagnosis is associated with postoperative outcome. Methods: In this prospective study, fine‐needle aspirate samples were obtained from 50 mammary tumors in 50 dogs. Results of cytologic and histopathologic examination were compared, using the histologic diagnosis as the reference method. Kaplan–Meier log rank analysis was used to evaluate univariate association of the cytologic diagnosis with duration of survival, local control, and metastasis‐free interval. Results: Adequate cytologic samples were obtained in 43/50 (86%) cases. The cytologic diagnosis correlated with the histologic diagnosis for benign and malignant tumors in 40/43 (93%) and 35/43 (81%) cases, respectively. Cytologic examination had a sensitivity of 88% and a specificity of 96% for the diagnosis of malignancy. The cytologic diagnosis had significant univariate association with duration of survival (P=.016), recurrence‐free interval (P=.003), and metastasis‐free interval (P=.014). Conclusions: Cytologic examination of mammary tumors in the dog has satisfactory accuracy, sensitivity, and specificity for the diagnosis of malignancy and is associated with postoperative outcome. Further studies on the diagnostic accuracy of cytology as well as multivariate analysis of its preoperative prognostic value in mammary tumors in the dog are warranted.     

监测饲料及其原料以消除霉菌毒素的影响

由于霉菌毒素是一类无色无臭无味的物质,所以测定饲料原料和饲料成品以监测霉菌毒素的污染是非常重要的. 霉菌毒素这一术语源自希腊语单词"mycos"和"toxikon",是指在各种各样环境条件下由真菌产生的次生有毒代谢产物.     

Computed tomographic features of incomplete ossification of the canine humeral condyle

Objectives— To describe computed tomographic (CT) features of canine elbows with incomplete ossification of the humeral condyle (IOHC) and investigate co-existing incongruence in the elbow joint.
Study Design— Case control study.
Animals— Dogs with IOHC (n=20; 38 elbows) and 25 normal elbows.
Methods— Elbows with IOHC and normal elbows were assessed by CT. Standardized dorsal and sagittal reconstructions were created at 3 levels using image analysis software to obtain single measurements of the humero-radial and humero-ulnar joint spaces. On dorsal plane reconstructions, joint space measurements were obtained at the center point of the humero-radial and humero-ulnar articulations. Joint incongruity was defined as the difference between the humero-radial and the humero-ulnar joint spaces.
Results— Nineteen dogs (95%), all Spaniel breeds, had either bilateral IOHC demonstrable as a saw-toothed intercondylar complete or incomplete hypoattenuating defect with hyperattenuating margins, or IOHC with contralateral humeral condylar fracture (HCF). Joint incongruity values for IOHC were compared with those of normal elbows. Significant differences were noted at the levels of the medial coronoid apex ( P <.0001) and base ( P <.004) indicative of humero-ulnar incongruence. Evidence of medial coronoid disease in 10 elbows (26%) and degenerative joint disease in 30 elbows (79%) was also found.
Conclusions— Presence of elbow incongruence may be an underlying factor in failure of ossification centers to fuse leading to IOHC.
Clinical Relevance— IOHC is clearly defined by CT, and it should be considered in larger Spaniel breeds, with a chronic forelimb lameness or HCF.     

Simulating the soil phosphorus dynamics of four long-term field experiments with a novel phosphorus model

Phosphorus is a nonrenewable resource, which is required for crop growth and to maintain high yields. The soil P cycle is very complex, and new model approaches can lead to a better understanding of those processes and further guide to research gaps. The objective of this study was to present a P-submodel, which has been integrated in the existing Carbon Candy Balance (CCB) model that already comprises a C and N module. The P-module is linked to the C mineralization and the associated C-pools via the C/P ratio of fresh organic material. Besides the organic P cycling, the module implies a plant-available P-pool (P_av), which is in a dynamic equilibrium with the nonavailable P-pool (P_na) that comprises the strongly sorbed and occluded P fraction. The model performance was tested and evaluated on four long-term field experiments with mineral P fertilization, farmyard manure as organic fertilizer and control plots without fertilization. The C dynamics and the P_av dynamics were modelled with overall good results. The relative RMSE for the C was below 10% for all treatments, while the relative RMSE for P_av was below 15% for most treatments. To accommodate for the rather small variety of available P-models, the presented CNP-model is designed for agricultural field sites with a relatively low data input, namely air temperature, precipitation, soil properties, yields and management practices. The CNP-model offers a low entry threshold model approach to predict the C-N and now the P dynamics of agricultural soils.     

Inverse Modeling to Quantify Irrigation System Characteristics and Operational Management

Remotely sensed (RS) data is a major source to obtain spatialdata required for hydrological models. The challenge for thefuture is to obtain besides the more direct observable data(landcover, leaf area index, digital elevation model andevapotranspiration), non-visible data such as soilcharacteristics, groundwater depth and irrigation practices.In this study we have explore the option of using inversemodeling to obtain these non-RS-visible data. For a commandarea in Haryana, India, we applied for the 2000–2001 rabiseason a RS-GIS-combined inverse modeling approach to derivenon-RS-visible data required in the regional application ofhydrological models. A Genetic Algorithm loaded stochasticphysically based soil-water-atmosphere-plant model (SWAP) wasdeveloped for the inverse problem and used in the study. Theresults showed good agreement with the inventoried data suchas soil hydraulic properties, sowing dates, groundwaterdepths, irrigation practices and water quality. The deriveddata could be used to predict the state of the system at anytime in the cropping season, which can be used to evaluateoperational management strategies.