Evaluation of Fertilizing Potential of Frozen-thawed dog Spermatozoa Diluted in ACP-106® using an In Vitro Sperm–Oocyte Interaction Assay

The aim of present study was to evaluate frozen canine semen with ACP-106 (Powder Coconut Water) using an in vitro sperm--oocyte interaction assay (SOIA). Ten ejaculates from five stud dogs were diluted in ACP-106 containing 20% egg yolk, submitted to cooling in a thermal box for 40 min and in a refrigerator for 30 min. After this period, a second dilution was performed using ACP-106 containing 20% egg yolk and 12% glycerol. Samples were thawed at 38 degrees C for 1 min. Post-thaw motility was evaluated by light microscopy and by using a computer aided semen analysis (CASA). Plasma membrane integrity and sperm morphology/acrosomal status were evaluated by fluorescent probes (C-FDA/PI) and Bengal Rose respectively. Moreover, frozen-thawed semen was analysed by a SOIA. Subjective post-thaw motility was 52.0 +/- 14.8% and it was significant higher than the total motility estimated by CASA (23.0 +/- 14.8%) because this system considered the egg yolk debris as immotile spermatozoa. Although normal sperm rate and acrosomal integrity evaluated by Bengal Rose stain was 89.6 +/- 3.1% and 94.3 +/- 3.1%, respectively, post-thaw percentage of intact plasma membrane was only 35.1 +/- 14.3%. Regarding SOIA, the percentage of interacted oocytes (bound, penetrated and bound and/or penetrated) was 75.3%. Using regression analysis, it was found significant relations between some CASA patterns and data for SOIA. In conclusion, the freezing-thawing procedure using ACP-106 was efficient for maintain the in vitro fertility potential of dog spermatozoa.     

Polymerase chain reaction and other laboratory techniques in the diagnosis of long incubation rabies in Australia

SUMMARY Blood and post-mortem tissues from a 10-years-old girl were submitted to the Australian Animal Health Laboratory. Clinical signs and histopathological lesions had suggested a diagnosis of rabies, but, an unusually long incubation period of at least 5 years did not encourage such a diagnosis. Serological examinations by the rapid fluorescent focus inhibition test revealed a dramatic increase in rabies virus-neutralising antibody during the 10-day period of hospitalisation. The results of a fluorescent antibody test on brain smears, and an immunoperoxidase test on formalin-fixed sections of brain were also consistent with a diagnosis of rabies. Attempts to isolate virus were unsuccessful. Polymerase chain reactions (PCRs) were conducted on a 10% suspension of a post-mortem sample from the patient's brain, using primers based on the published sequence of the Pasteur virus strain of rabies virus. 413 and 513 bp fragments from the nucleoprotein gene and a 403 bp fragment from the glycoprotein gene were amplified. Subsequent sequencing of these fragments, and comparison with equivalent regions of known rabies viruses, confirmed that the fragments originated from a virus belonging to the rabies virus serotype. This case demonstrated the advantage of using a range of laboratory techniques to obtain a definitive diagnosis. In particular, a PCR-based test may allow a diagnosis, even in the face of conditions that preclude virus isolation such as apparently occurred in this case. Finally, this case demonstrated that an unusually long incubation period should not discourage a tentative clinical diagnosis of rabies.     

Qualitative and quantitative ultrasound attributes of maternal‐foetal structures in pregnant ewes

The aim of this study was to examine foetal organs and placental tissue to establish a correlation between the changes in the composition of these structures associated with their maturation and the ultrasonographic characteristics of the images. Twenty‐four pregnant ewes were included in the study. Ultrasonography assessments were performed in B‐mode, from the ninth gestational week until parturition. The lungs, liver and kidneys of foetuses and placentomes were located in transverse and longitudinal sections to evaluate the echogenicity (hypoechoic, isoechoic, hyperechoic or mixed) and echotexture (homogeneous and heterogeneous) of the tissues of interest. For quantitative evaluation of the ultrasonographic characteristics, it was performed a computerized image analysis using a commercial software (Image ProPlus^®). Mean numerical pixel values (NPVs), pixel heterogeneity (standard deviation of NPVs) and minimum and maximum pixel values were measured by selecting five circular regions of interest in each assessed tissue. All evaluated tissues presented significant variations in the NPVs, except for the liver. Pulmonary NPVmean, NPVmin and NPVmax decreased gradually through gestational weeks. The renal parameters gradually decreased with the advancement of the gestational weeks until the 17th week and later stabilized. The placentome NPVmean, NPVmin and NPVmax decreased gradually over the course of weeks. The hepatic tissue did not show echogenicity and echotexture variations and presented medium echogenicity and homogeneous echotexture throughout the experimental period. It was concluded that pixels numerical evaluation of maternal‐foetal tissues was applicable and allowed the identification of quantitative ultrasonographic characteristics showing changes in echogenicity related to gestational age.     

Factors affecting the breeding success of the African Black Oystercatcher (Haematopus moquini): a perspective on protection and food availability

Breeding success (fledglings pair^?1 y^?1) of the Red-listed African Black Oystercatcher (Haematopus moquini) is highly variable, both spatially and temporally. Despite a diversity of natural factors causing this variability, there is evidence that two anthropogenic factors, i.e. disturbance and an introduced mussel (Mytilus galloprovincialis), are having an impact on the local breeding success of this species. Using a data set comprising 87 site-years of nest-monitoring data across most of the species’ breeding range, we analysed the extent and causes of variability in breeding success. Breeding success differed across three population categories defined by varying levels of human disturbance: island populations, protected mainland populations, and unprotected mainland populations. Differences in breeding success between island populations and protected mainland populations were likely due to differing exposure to predators; however, differences between protected and unprotected mainland populations were unlikely caused by this as both experience equivalent predation levels (although from different predators). Protection only improved the breeding success of oystercatchers in very high-quality habitats (with a high biomass of alien mussels), and where populations were ‘released’ from high levels of human disturbance. In unprotected mainland areas, human activity impacted on the breeding success of local populations primarily through predation of small chicks by uncontrolled dogs, and by rising tides drowning chicks that were hiding from human disturbance. The findings of this study note the potential conservation dilemma resulting from an invasive species improving the conservation status of a Red-listed species, and encourage the implementation of restricted sites in high-quality habitats with high breeding pair densities.     

Do subcutaneous sutures increase risk of laparotomy wound suppuration?

REASONS FOR PERFORMING STUDY: Incisional drainage and suppuration occurs commonly following exploratory laparotomy; any technique with the potential to reduce the incidence of this complication warrants investigation. OBJECTIVES: To determine if abandoning the use of subcutaneous sutures in laparotomy wound closure is safe and whether it reduces the risk of suppuration. METHODS: A randomised controlled study was carried out at 2 referral hospitals in the UK, involving 309 horses undergoing exploratory laparotomy; 150 horses underwent '2-layer' closure without a subcutaneous suture, while 159 underwent conventional '3-layer' closure. Information regarding degree of oedema and gross types of discharge was recorded daily; suppuration was defined as discharge of pus. Telephone follow-up was carried out 30 days after hospital discharge to identify those complications occurring after that date and, thereafter, every 3 months. The influence of closure method on risk of wound suppuration was assessed by Chi-squared analysis and by logistic regression. Time to suppuration was modelled using a Cox proportional hazards model. RESULTS: No catastrophic failures of 2-layer closures were recorded. Prevalence of suppuration was not significantly different, being 18.7% and 23.9% for 2- and 3-layer closures, respectively (OR = 1.37, 0.79-2.37, P = 0.263). CONCLUSIONS AND POTENTIAL RELEVANCE: This study found no significant difference in prevalence or rate of wound suppuration in 2-layer closures compared to conventional 3-layer closure. Two-layer closure is recommended as a safe alternative means of achieving ventral midline abdominal closure in horses.     

Epidemic of blindness in kangaroos - evidence of a viral aetiology

OBJECTIVE: To determine the cause of an epidemic of blindness in kangaroos. DESIGN AND PROCEDURES: Laboratory examinations were made of eyes and brains of a large number of kangaroos using serological, virological, histopathological, electron microscopical, immunohistochemical methods, and PCR with cDNA sequencing. In addition, potential insect viral vectors identified during the disease outbreak were examined for specific viral genomic sequences. SAMPLE POPULATION: For histopathological analysis, 55 apparently blind and 18 apparently normal wild kangaroos and wallabies were obtained from New South Wales, Victoria, South Australia, and Western Australia. A total of 437 wild kangaroos and wallabies (including 23 animals with apparent blindness) were examined serologically. RESULTS: Orbiviruses of the Wallal and Warrego serogroups were isolated from kangaroos affected with blindness in a major epidemic in south-eastern Australia in 1994 and 1995 and extending to Western Australia in 1995/96. Histopathological examinations showed severe degeneration and inflammation in the eyes, and mild inflammation in the brains. In affected retinas, Wallal virus antigen was detected by immunohistochemical analysis and orbiviruses were seen in electron microscopy. There was serological variation in the newly isolated Wallal virus from archival Wallal virus that had been isolated in northern Australia. There were also variations of up to 20% in genotype sequence from the reference archival virus. Polymerase chain reactions showed that Wallal virus was present during the epidemic in three species of midges, Culicoides austropalpalis, C dycei and C marksi. Wallal virus nucleic acid was also detected by PCR in a paraffin-embedded retina taken from a blind kangaroo in 1975. CONCLUSION: Wallal virus and perhaps also Warrego virus are the cause of the outbreak of blindness in kangaroos. Other viruses may also be involved, but the evidence in this paper indicates a variant of Wallal virus, an orbivirus transmitted by midges, has the strongest aetiological association, and immunohistochemical analysis implicates it as the most damaging factor in the affected eyes.     

Interactions Among Different Devices and Electrical Stimulus on the Electroejaculation of Captive Agoutis (Dasyprocta leporina)

The interactions among different electroejaculation devices associated with serial or continuous stimuli were investigated to improve the efficiency of the electroejaculation for semen collection in agoutis. Ten sexually matured male Dasyprocta leporina were restrained by the intramuscular administration of xylazine–ketamine association. Each individual was randomly subjected to four electroejaculation protocols, by combining two devices (one presenting longitudinal electrodes emitting square waves and other presenting ring electrodes emitting sine waves) and two electrical stimuli protocols (serial or continuous). A total of 40 attempts for electroejaculation were conducted in agoutis, being 10 per treatment. The most efficient treatment in providing ejaculates containing sperm (p < 0.05) was that using and electroejaculator connected to a probe with ring electrodes and associated with serial stimuli (4/7; 57%). In spite of semen parameters obtained by sine waves were adequate for using the samples for assisted reproduction, higher values for sperm motility and functional membrane integrity were obtained in the use of the square wave, independently of the electric stimulation protocol used (p < 0.05). In conclusion, we verified that the use of a device presenting a probe with ring electrodes and emitting sine waves, associated with a serial stimuli protocol, improves the efficiency for semen obtaining by electroejaculation in adults D. leporina.     

Fusion podoplasty for the management of chronic pedal conditions in seven dogs and one cat

Eight animals underwent fusion podoplasties for the treatment of chronic interdigital furunculosis (n=3), ectrodactyly (n=1), digit abnormalities associated with tendonectomy (n=1), redundant indertigital skin (n=1), conformational deformity (n=1), and necrotizing fasciitis of the paw (n=1). Median duration of bandaging was 14 days, and median duration of hospitalization was 5 days. Four dogs had dehiscence, which occurred at a mean time of 11 days after surgery. Clinical abnormalities necessitating podoplasty resolved in six animals and improved in two. Six animals had normal ambulation and two dogs had slight weight-bearing lameness after a median follow-up time of 29 mo. Fusion podoplasty may be recommended as a salvage procedure for the treatment of various chronic pedal diseases in dogs and cats.