Ultrastructural Characteristics of Non-matured and In Vitro Matured Oocytes Collected from Pre-pubertal and Adult Domestic Cat Ovaries

The present study describes the ultrastructural characteristics of cat oocytes before maturation and after 12- and 24-h in vitro maturation (IVM). Oocytes were recovered from pre-pubertal and adult queen ovaries after ovariohysterectomy and a proportion were stored in glutaraldehyde at 4°C until examination by transmission electronic microscopy (TEM). Those selected for maturation were cultured before TEM in DMEM for 12 and 24 h at 38°C in a humidified environment of 5% O₂, 5% CO₂ and 90% N₂. Specimens were divided into six groups: non-matured oocytes from pre-pubertal queens (PP0), non-matured oocytes from adult queens (A0), 12-h in vitro matured oocytes from pre-pubertal queens (PP12), 12-h in vitro matured oocytes from adult queens (A12), 24-h in vitro matured oocytes from pre-pubertal queens (PP24) and 24-h in vitro matured oocytes from adult queens (A24). Across the treatment groups, it was possible to observe differences in the thickness of the perivitelline space, the penetration of cumulus cell projections forming a junctional complex, distribution and density of small vesicles, lipid droplets, microvilli, mitochondria and cortical granules and variable degrees of development of Golgi complexes. These findings demonstrated that ultrastructural analysis of oocytes matured in vitro is a valuable tool to evaluate oocyte cytoplasmic maturation and that this IVM protocol was efficient in inducing gradual morphological changes necessary for cytoplasmic maturation of pre-pubertal and adult cat oocytes.     

Surgical Treatment of a Canine Intranasal Meningoencephalocele

Objective— To report the clinical signs, diagnosis, and surgical treatment of an intranasal meningoencephalocele in a dog.
Study Design— Case report.
Animal— Female Border collie, 5 months old.
Methods— A right intranasal meningoencephalocele was identified by computed tomography and magnetic resonance imaging.
Results— The lesion was approached by a modified transfrontal craniotomy. Surgical closure of the defect at the level of the cribriform plate and removal of extruded brain tissue resulted in regression of lacrimation and coincided with absence of seizuring. Treatment with phenobarbital was gradually reduced and stopped at 7 months after surgery. At 28 months the dog remained free of seizures.
Conclusion— Meningoencephalocele, although rare, can cause seizures in dogs and can be treated surgically.
Clinical Relevance— A transfrontal craniotomy with excision of the meningoencephalocele and closure of the defect can be an effective treatment for an intranasal meningoencephalocele in dogs.     

Occurrence of Trypanosoma evansi in Horses in the State of Minas Gerais,Brazil

Ttrypanosomosis caused by Trypanosoma evansi is a condition that causes significant losses to farmers in endemic areas. This study aimed to report one case of trypanosomiasis in the municipality of Itabira, state of Minas Gerais, Brazil. In November 2010, on a farm with 16 horses, three horses had clinical signs of anemia, limb edema, weight loss, lameness, muscle atrophy, and incoordination of the hind limbs. Trypanosomosis was suspected, and the animals with clinical signs were treated with two doses of diminazene aceturate (7 mg/kg, intramuscular) every 7 days; the treated animals recovered from the clinical symptoms. Blood samples were collected from six horses on the farm, including the three treated animals, to perform polymerase chain reaction specific for T evansi. Four samples were polymerase chain reaction negative, including those collected from three treated horses. However, two other asymptomatic horses were positive for the parasite based on the molecular testing. Based on the results, we concluded that the initial clinical suspect of trypanosomosis was correct, and the treatment used was effective. Probably the disease was introduced to Minas Gerais State through a stallion, which acquired the infection in an endemic area in southern Brazil.     

Prophylactic treatment with flumethrin,a pyrethroid (Bayticol?, Bayer), against Anaplasma phagocytophilum infection in lambs

Backgroud

Anaplasma phagocytophilum (formerly Ehrlichia phagocytophila) causes the disease tick-borne fever (TBF) in domestic ruminants and has for decades been one of the main scourges for the sheep industry in the coastal areas of Norway. Current control strategies are based on reduction of tick infestation by chemical acaricides.

Methods

In the present study, we investigated if frequent pour-on applications of pyrethroids would reduce tick infestion rate and seroprevalence of A. phagocytophilum infection in sheep. Forty lambs, one month old, of the Norwegian White Sheep breed were used. The lambs belonged to the experimental sheep flock at the Department of Production Animal Clinical Sciences. None of the lambs had been on I. ricinus infested pasture before turnout (day 0). All lambs were twins and twenty lambs were treated with a pour-on pyrethroid (Bayticol®, Bayer A/S, DK-2300) with a dose of 5 ml on days 0, 14, 28, 42, 56, 70, 84, 98, 112 and 128. Twenty lambs were untreated controls. The lambs were collected every fourteen days on pasture for treatment. In addition, the lambs were examined for ticks, blood sampled, weighed, and rectal temperature was recorded.

Results and conclusion

A significant reduction in tick infestion rate was detected on treated lambs. However, the present results indicate that frequent acaricide treatment does not reduce the seroprevalence to A. phagocytophilum on tick-infested pasture.     

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.     

Basic reproduction number for equine-2 influenza virus a (H3N8) epidemic in racehorse facilities in Japan, 1971

Characterizing the epidemiologic determinants of equine influenza to facilitate effective control measures, including determining the critical proportion of the population that must be immunized to achieve containment, is critical. Through the use of counting process and martingale method, the basic reproduction number (R₀; a measure of transmission potential) of equine-2 influenza A virus (H3N8) infection was estimated based on outbreak data recorded in 1971. The final attack rate ranged from 81.9% to 99.4%. Our estimate of R₀ was in the order of 2 to 5, which was smaller than the previously suggested estimate. Numerical analysis supported airborne spread as the major mode of transmission. Given that simple threshold vaccination coverage might not be realistic within racing stables, the combination of movement restrictions, isolation, and other countermeasures alongside vaccination is recommended to contain outbreaks of equine influenza.     

Effects of a Specific Thromboxane Synthetase Inhibitor on Development of Experimental Dirofilaria immitis Immune Complex Glomerulonephritis in the Dog

Twelve Beagle dogs were immunized with aqueous-soluble Dirofilaria immitis antigens, and subsequent to at least fivefold increases in serum antibody titer, 6 mg of homologous antigen was infused into the left renal artery. Six dogs were treated once daily starting the day of infusion with 0.75 mg/kg of 1-benzylimidazole (1-BIM) in saline. Six control dogs were given saline only. Light, immunofluorescent, and transmission electron microscopic examinations of renal tissue from control dogs, 10 days after antigen infusion, showed a mesangioproliferative glomerulonephritis in the left kidney with polymorphonuclear leukocyte (PMNL) infiltration and fibrin deposition. Immunoglobulin (Ig) G, M, C3, and Dirofilaria antigen deposits were observed in a segmental granular pattern. Mesangial, subendothelial, and intramembranous electron dense deposits were observed, and anti-Dirofilaria antibodies were demonstrated in kidney eluates from each dog. Administration of 1-BIM had no significant effect on IgG, IgM, C3, or antigen deposits, electron dense deposits, or concentration of antibody in kidney eluates. However, 1-BIM-treated dogs had less glomerular cell proliferation, periodic acid-Schiff (PAS) positive glomerular staining, PMNL infiltration, and fibrin deposition. These data suggest that thromboxane is an important mediator in the development of immune complex glomerulonephritis, and that in certain circumstances, inhibition of thromboxane synthesis may be an effective therapy for immune complex glomerulonephritis in the dog.     

Electrocoagulative removal of endometrial cysts in the mare

Six mares had endometrial cysts removed by endoscopic electrocoagulative surgery. These mares exhibited poor reproductive histories and endometrial cysts were the only detectable abnormalities. In each case, at least two cysts greater than 2.5cm were present. One mare subsequently produced a live foal; two are presently in foal; one conceived but absorbed; one formed more cysts after surgery; and one has not yet been bred. Histologically, the walls of the cysts contained entire lamina propria, relatively normal endometrial glands, and small cystic spaces.