Genetic characterisation of oak seedlings, epicormic, crown and micropropagated shoots from mature trees by RAPD and microsatellite PCR

DNA was isolated from seedlings of Quercus robur, collected from a single provenance, and from epicormic, crown shoots and in vitro shoots from a single tree of Q. petraea using a CTAB method of extraction. DNA was obtained in sufficient quantity and purity, from 13 out of 30 seedlings, and from all isolations from epicormic and in vitro shoots (2.5–10.0 μg/g fresh/ weight). Smearing was minimised at a primer concentration of 0.12 μM with Taq polymerase at 0.5 unit/reaction. Nine primers produced 142 bands, 28 of which were polymorphic. A similarity index showed that 11 seedlings were closely related with high coefficients (0.85–0.90), but each could be identified from another using only 9 primers (OPA-02 and -05, OPG-04 and -05, OPE-01, -02, -03, -08, -09). DNA was isolated from crown, epicormic and in vitro leaves originating from a single 150-yr old tree of Q. petraea and analysed by randomly amplified polymorphic DNA (RAPD) and microsatellites. With each primer, a characteristic RAPD pattern was obtained, and it was common to all six epicormic shoots derived from different parts of a single branch of this tree; also to the shoots from the crown of the same tree with OPE1 OPA-05, OPA-08, OPA-01, OPA-02, OPA-04, OPA-05, OPG-02, OPG-10, OPE-12. Similarly, the RAPD pattern obtained from shoot cultures in vitro, derived from individual nodes of epicormic shoots produced by six different branch segments, were uniform for each of 15 primers. This work was repeated using microsatellite PCR. Three microsatellite loci AG16, AG 1/2 and AG 1/5 were amplified by PCR. It showed a uniformity of these microsatellite loci in shoots from the crown of the tree, and from epicormic shoots cultures derived from six different sections of branch.     

RADIOGRAPHIC CHARACTERIZATION OF PRIMARY LUNG TUMORS IN 74 DOGS

Primary pulmonary neoplasia is well recognized in dogs and prognosis depends upon the tumor type. The purpose of this retrospective study was to characterize the radiographic appearance of different primary lung tumors with the goal of establishing imaging criteria to separate the different types. Three‐view thoracic radiographs of 74 dogs with histologically confirmed pulmonary anaplastic carcinoma (n = 2), adenocarcinoma (n = 31), bronchioalveolar carcinoma (n = 19), histiocytic sarcoma (n = 21), and squamous cell carcinoma (n = 1) were evaluated. Radiographs were assessed for tumor volume, affected lobe, location within lobe, overall pulmonary pattern, presence of cavitation, mineralization, air bronchograms, lymphadenomegaly, and pleural fluid. Histiocytic sarcomas were significantly larger than other tumor types (271 cm³; P = 0.009) and most likely to be found in the left cranial (38%; 8/21) and right middle (43%; 9/21) lung lobes, whereas adenocarcinomas were most likely to be found in the left caudal (29%; 9/31) lung lobe. Fifty‐seven percent (12/21) of histiocytic sarcomas had an internal air bronchogram. Findings indicate that a large mass in the periphery or affecting the whole lobe of the right middle or left cranial lung lobe with an internal air bronchogram is likely to be an histiocytic sarcoma.     

Clinical and pharmacokinetic characteristics of intracavitary administration of pegylated liposomal encapsulated doxorubicin in dogs with splenic hemangiosarcoma

BACKGROUND: Canine splenic hemangiosarcoma (HSA) is a fatal malignancy, and most affected dogs die within a few months of diagnosis. Most dogs present with signs from tumor rupture, resulting in hemoabdomen and intra-abdominal dissemination. The abdomen is also the main site of disease recurrence. HYPOTHESIS: Intraperitoneal (IP) administration of doxorubicin will delay or prevent intra-abdominal tumor recurrence and prolong survival in dogs with HSA. ANIMALS: Fourteen dogs with splenic HSA. METHODS: A prospective, unmasked, uncontrolled clinical trial. After staging of disease status and splenectomy, pegylated liposomal encapsulated doxorubicin was administered intraperitoneally (1 mg/kg body weight) every 3 weeks for 4 cycles. All dogs were monitored for recurrence of HSA. Samples of plasma and abdominal fluid were collected for measurement of doxorubicin concentration and pharmacokinetic analysis. Nonlinear mixed-effect modeling was used to describe the pharmacokinetics of liposomal doxorubicin administered IP. RESULTS: All 14 dogs died, 12 because of HSA and 2 from other causes. Postmortem examination was performed on 12 dogs. All 12 dogs died because of HSA-related causes and had hepatic metastases and hemoabdomen. The IP-treated dogs had fewer serosal, mesenteric, and omental metastases than historical controls treated with systemic doxorubicin. Results of the postmortem examination and pharmacokinetic analysis confirmed that IP delivery of doxorubicin resulted in an effective drug concentration with a clearance comparable with that after i.v. delivery. CONCLUSIONS AND CLINICAL IMPORTANCE: IP pegylated liposomal encapsulated doxorubicin administration did not prevent intraabdominal recurrence of HSA in dogs.     

The interaction of Cytolin and NAA on cropping red ‘Delicious’ apple

A trial, at Spreyton in north-western Tasmania, studied the effects of Cytolin and NAA sprays on the thinning and shape of ‘Delicious’ apples. Untreated control and hand thinned trees were compared with trees treated with Cytolin or NAA alone, or in combination. Cytolin was applied at 25 mg 1~‘ at the king petal (KP) stage. NAA was applied at 8 mg I“1 at full bloom (FB) and 7,10, or 15 days after full bloom (AFB). Cytolin alone had no effect on thinning, fruit weight or on pip numbers but it did improve fruit shape. The combination of cytolin and NAA at FB overthinned. Cytolin and NAA combinations applied at all times after FB significantly decreased both mean fruit weight and pip numbers and reduced the percentage of fruit > 70 mm. All Cytolin-NAA combinations and the NAA treatments applied at 10 and 15 days AFB resulted in unacceptably high percentages of apples < 45 mm in diameter (pygmy fruit). It is recommended that Cytolin and NAA should not be used in the same spray programme. NAA alone is a most effective thinner at FB or seven days AFB.     

Diagnosis of lumbosacral intervertebral disc disease and protrusion in a horse using ultrasonographic evaluation and computed tomography

This case report describes the clinical presentation, diagnostic imaging modalities, treatment and post mortem evaluation of lumbosacral intervertebral disc protrusion in a mature Quarter Horse gelding 10 days after initial signs were noted. Grade 3 hindlimb ataxia, conscious proprioceptive deficits, urinary and faecal incontinence were present, which did not improve with anti‐inflammatories, antimicrobial therapy, corticosteroids, antioxidant therapy, cold‐laser therapy or electroacupuncture. Imaging modalities utilised ante mortem were computed radiography, transcutaneous and transrectal ultrasonography. Transrectal ultrasonography yielded findings highly suggestive of lumbosacral intervertebral disc protrusion and due to the lack of improvement and a poor prognosis, the horse was humanely subjected to euthanasia. Post mortem computed tomography, necropsy and histopathological evaluation confirmed lumbosacral intervertebral disc disease and protrusion into the spinal canal with subsequent impingement of the spinal nerve roots. Lumbosacral intervertebral disc protrusion as a clinical disease in the horse has not been previously described and should be included as a differential diagnosis in cases with acute hindlimb ataxia, proprioceptive deficits, and urinary and faecal incontinence.     

Effect of an Otic Milbemycin Oxime Formulation on Tegastes acroporanus Infesting Corals

The copepod Tegastes acroporanus is a notorious pest of captive corals in the genus Acropora. In recent years, infestations of T. acroporanus have become widespread among public aquaria and coral propagation facilities and have been largely controlled with the extra-label use of milbemycin oxime formulations (Carl 2008). Many of these drug formulations (which were intended for dogs) have been discontinued by their manufacturers in favor of multidrug products, many of which are unsuitable for corals, forcing experimentation with alternatives. This report provides the first data on populations of T. acroporanus treated with milbemycin oxime and documents the first known use of an otic solution, MilbeMite Otic (Novartis Animal Health U.S., Greensboro, North Carolina), against copepods on live corals. MilbeMite Otic was found to be soluble in seawater and successful at eradicating T. acroporanus in a large exhibit over the course of 6-h waterborne baths (n = 12) at 0.167 µg/L. The resident population of T. acroporanus was also quantified before each treatment to provide the first estimates of coral parasite burden in response to the application of a waterborne chemotherapeutic agent.

Received November 19, 2015; accepted June 7, 2016 Published online October 24, 2016     

Where art thou louse? A snapshot of attachment location preferences in salmon lice on Atlantic salmon hosts in sea cages

Problematic sea lice infestations on farmed Atlantic salmon (Salmo salar) have motivated extensive research and development into new methods to prevent, monitor and control sea lice. Most of these technologies require detailed information on the behaviour, spatial distribution and demography of lice on host fish. This study investigated how salmon lice (Lepeophtheirus salmonis) infestation density varies across the host's surface under sea cage farming conditions. Lice abundance, demography and attachment location were tracked over time, with repeated sampling of 300 individually tagged salmon across three replicate experimental sea cages. The data reveal clear differences in attachment locations according to sex and stage, but with an overall preference for the dorsal surface among mobile stages—dorsal head for adult females and dorsal-posterior section for males and pre-adults. Total lice abundance was highly variable between repeated measures of individual fish, consistent with frequent host-switching or mortality. Total lice numbers also declined between sampling dates, likely due to handling, with lost mobile lice being almost exclusively adult males. As the distribution of sea lice on hosts is likely determined by numerous factors, future image-based automated detection systems should be validated in settings that reflect the complex host–parasite interactions that occur in open farming systems.