Expression of Orexin A and its Receptor 1 in the Epididymis of the South American Camelid Alpaca (Vicugna pacos)

Orexins A (ox A) and B are two peptides originally discovered in neurons of rat hypothalamus, and later found in different cellular types of the gastrointestinal and genital tracts. They arise from the proteolytic cleavage of a common precursor molecule, prepro‐orexin, and bind to two receptors, namely receptor 1 (ox1r) and receptor 2 for orexins, that show different binding affinity. The central role of the two peptides has been extensively studied, whereas their activity in the periphery is still poorly known. Here, we investigated the presence of ox A and ox1r in the epididymis of a South American camelid species, the alpaca, by immunohistochemistry, and we also assessed the expression of prepro‐orexin and ox1r in tissue extracts by Western blotting analysis. Ox A‐ and ox1r‐immunoreactivity was found in the cytoplasm of principal cells of the caput epididymis. A prevalent supranuclear localization of granular‐shaped positive material was observed. No positivity was present in the other cytotypes of epididymis. The expression of two peptides with molecular weight corresponding to those of prepro‐orexin and ox1r, respectively, was detected in the tissue extracts from the organ.     

FEASIBILITY FOR USING HYPOFRACTIONATED STEREOTACTIC VOLUMETRIC MODULATED ARC RADIOTHERAPY (VMAT) WITH ADAPTIVE PLANNING FOR TREATMENT OF THYMOMA IN RABBITS: 15 CASES

Thymoma is a relatively common tumor in rabbits. Treatment with surgery, radiation therapy, and chemotherapy alone or in combination has been reported with varying outcomes. Stereotactic volumetric modulated arc radiotherapy delivered in a hypofractionated manner allows high doses of radiation to be delivered to the target volume while allowing sparing of adjacent critical structures. This therapy is ideally suited for thymomas in rabbits given their size, the difficulty of multiple anesthesia episodes and the complexity of the radiotherapy plans required due to the tumor's proximity to the heart, lungs, and mediastinal structures. Fifteen rabbits with thymoma were prospectively recruited for this observational, single institution, single arm clinical study. All rabbits were imaged with both computed tomography (CT) and magnetic resonance imaging (MRI). A total dose of 40 Gy in six fractions was delivered using a single arc over an 11‐day period with repeat CT simulation done every other fraction for adaptive planning. Follow‐up evaluation was done through repeat CT and MRI imaging and revealed complete responses using the Response Evaluation Criteria in Solid Tumors (RECIST) criteria. Two rabbits had died at 618 and 718 days, 10 were alive and three were lost to follow‐up. Observed acute and late effects were graded according to the Veterinary Radiation Therapy Oncology Group (VRTOG) criteria and were found to be minimal.     

Length–mass allometries in amphibians

Body mass is rarely recorded in amphibians, and other body measurements (e.g. snout to vent length, SVL) are generally collected instead. However, length measurements, when used as proxies of body mass in comparative analyses, are problematic if different taxa and morphotypes are included. We developed allometric relationships to derive body mass from SVL measurements. We fitted phylogenetic generalized least square models for frogs (Anura) and salamanders (Caudata) and for several families separately. We tested whether allometric relationships differed between species with different habitat preferences and between morphs in salamanders. Models were fitted with SVL–mass measurements for 88 frog and 42 salamander species. We assessed the predictive performance of the models by cross‐validation. Overall, the models showed high explained variance and low forecasting errors. Models differed among semi‐aquatic, terrestrial and arboreal frogs, and between paedomorphic and non‐paedomorphic salamanders. Body mass estimates derived from our models allow for comparability of studies on multiple taxa and can be used for testing theories built upon evolutionary and ecological processes which are directly related to body mass.     

Immunohistochemical detection of P-glycoprotein (clone C494) in canine mammary gland tumours

Elevated levels of P-glycoprotein have been reported in multidrug-resistant tumours in both humans and dogs. In the present study, we investigated the expression of P-glycoprotein in 57 canine mammary gland tumours, 10 mammary gland hyperplasia and seven normal mammary glands by immunohistochemistry. Tissue sections were incubated with an anti-Pgp monoclonal antibody and visualized with En Vision-DAB polymer. Normal and hyperplastic mammary tissues were negative or showed slight cytoplasmic immunoreactivity. Neoplastic cells in benign mammary tumours showed diffuse cytoplasmic staining, in contrast to malignant tumours that showed mainly a membranous staining pattern for Pgp (C494). We observed statistically significant differences among all the different groups of tissues analysed except for benign tumours versus hyperplasia (P = 0.221). Receiver-operating characteristic analysis showed that the best cut-off point to differentiate the threshold to differentiate negative from positive tissue samples was 18.40% of immunostained cells. These results provide a first indication that routine evaluation of Pgp expression in canine mammary gland tumours, taking into consideration a cut-off point for positivity, may be useful for selecting cases for chemotherapy.     

Monitoring of antimicrobial resistance in healthy dogs: first report of canine ampicillin-resistant Enterococcus faecium clonal complex 17

National surveillance programs on antimicrobial usage and antimicrobial resistance in animals have been established in various countries but few of them include bacteria from pets. The objectives of this study were to assess the prevalence of antimicrobial resistance in healthy dogs and to search for resistance phenotypes of clinical relevance. Escherichia coli and Enterococcus spp. were isolated from faecal swabs obtained from 127 dogs. Disk diffusion was used to measure antimicrobial susceptibility in 117 Escherichia coli, 10 Enterococcus faecium and 51 Enterococcus faecalis of canine origin. Resistance was relatively low compared with food animal species in Denmark. All Escherichia coli isolates were susceptible to broad-spectrum aminopenicillins, third generation cephalosporins and fluoroquinolones. Despite the low prevalence of resistance, statistical analysis of questionnaire data revealed a significant association (p=0.02) between recent antimicrobial treatment and resistance in Escherichia coli. Interestingly, two dogs were found to shed Enterococcus faecium resistant to ampicillin. Multilocus sequence typing of these isolates indicated that the two isolates belonged to sequence types associated with human nosocomial infections, and one (ST-192) was genetically related to human epidemic clonal complex 17. The detection of ampicillin-resistant Enterococcus faecium warrants further studies on the prevalence of these bacteria in dogs and on the possible implications to both animal and human health. The results suggest that distinct methods for detection and assessment of antimicrobial resistance in animals should be considered depending on the target animal species and the purposes of the study.     

Cat and dog ownership and management patterns in central Italy

Three hundred and ninety-seven randomly selected households were interviewed by telephone to determine the numbers and management of owned cats and dogs in the Teramo Province of Italy. The households were selected using stratified random sampling for each municipality; municipalities were combined into coastal, central hills and mountain regions for analysis. The interviews were completed during May and June of 2004 with a response rate of 74% (397/536). Forty-six percent of households (n=181) owned pets; 15% of all households (n=60) owned cats and 33% (n=130) owned dogs. Twenty-seven of these households (7%) owned both cats and dogs. Data were provided on 91 cats evenly divided between males and females. The median age was 3 years (range 0.2-10 years). Forty-one percent of cats (36/87) entered the household as strays. Nearly half lived entirely outside. Seventy percent (62/88) had visited a veterinarian at least once; 43% (39/91) were sterilized. About 1/3 had had a litter and all litters were considered accidental rather than planned. Age, indoor/outdoor status, veterinarian visit and region were all associated with sterilization. Age, confined to a yard, veterinary visit and region were associated with allowing the cat to roam freely. Data were provided on 182 dogs. Sixty-two percent (113/181) were male, with a median age of 4 years (range newborn to 17 years) and 40% (72/181) were purebred. Almost half were acquired as a gift. Sixty-two percent (112/180) were kept entirely outside despite the fact that 82% (147/180) were considered companions rather than working dogs. Almost all of the dogs had been to a veterinarian at least once; only 20% (n=29) were sterilized. Male dogs were significantly less likely to be sterilized than females. Almost half the dogs had had at least one litter. Seventy-six percent (137/180) of dogs knew some basic commands. Sex, source and training to sit/stay/come were significantly associated with whether the dog was sterilized. Dog size, confinement to a yard, trained to leash walk, veterinary visit and region were predictors of roaming freely. Source and trained to leash walk were associated with dog registration. For cats and dogs, education about sterilization seemed to be critically important. For dogs, factors relating to training, which may reflect the strength of the human-animal bond, were also important.     

Definition and clinical evaluation of a recruiting airway pressure based on the specific lung elastance in anesthetized dogs

ObjectiveTo determine the specific lung elastance (SE_L) in anesthetized dogs and to evaluate the efficacy of a SE_L-based recruiting airway pressure (RP_aw) at improving global and regional lung aeration.Study designRetrospective and prospective clinical study.AnimalsA total of 28 adult dogs were included in the retrospective study and six adult dogs in the prospective study.MethodsRetrospective study: SE_L and SE_L-based RP_aw were determined using previously published data. In mechanically ventilated dogs undergoing thoracic computed tomography (CT), SE_L was calculated as ΔP_L/(V_T/EELV), where ΔP_L is the driving transpulmonary pressure, V_T is the tidal volume and EELV is the end-expiratory lung volume. The ratio of lung to respiratory system elastance (E_L/E_rs) was determined. SE_L and E_L/E_rs were used to calculate the SE_L-based RP_aw. Prospective study: dogs underwent thoracic CT at end-expiration and at end-inspiration using the SE_L-based RP_aw, and global and regional aeration was determined. For analysis of regional aeration, lungs were divided into cranial, intermediate and caudal regions. Regional compliance was also calculated. A p value <0.05 was considered significant.ResultsThe SE_L and E_L/E_rs were 12.7 ± 3.1 cmH₂O and 0.54 ± 0.07, respectively. The SE_L-based RP_aw was 29.1 ± 7.6 cmH₂O. In the prospective study, the RP_aw was 28.2 ± 1.3 cmH₂O. During RP_aw, hyperinflation increased (p = 0.0003) whereas poorly aerated (p < 0.0001) and nonaerated (p = 0.01) tissue decreased. Normally aerated tissue did not change (p = 0.265). Regional compliance was higher in the intermediate (p = 0.0003) and caudal (p = 0.034) regions compared with the cranial region. Aeration did not differ between regions (p > 0.05).Conclusions and clinical relevanceAn SE_L-based RP_aw reduces poorly and nonaerated lung tissue in anesthetized dogs. In nonsurgical anesthetized dogs, an RP_aw near 30 cmH₂O is effective at improving lung aeration.