The Acidic Probe LysoSensor™ is not Useful for Acrosome Evaluation of Cryopreserved Ram Spermatozoa

To try new acrosomal probes for assessing ram spermatozoa, we compared the LysoSensor^? probe, which labels acidic organelles, with the frequently used peanut agglutinin acrosomal probe (PNA‐PE; phycoerythrin as fluorescent moiety). The previous microscopic observations showed a lack of relationship of LysoSensor^? with acrosomal status. Semen was obtained from five rams and frozen in four pools. Each pool was analysed carrying out a triple staining propidium ioide/PNA‐PE/LysoSensor^? Green DND‐189 to test acrosome labelling, and a double staining SYBR‐14/PI, to assess sperm viability. Stained samples were analysed by flow cytometry. All measurements were replicated. Data were processed using agreement and repeatability tests. LysoSensor^? labelling did not agree with PNA (mean of differences: 30.8%; coefficient of agreement: 22.6%), confirming microscopic observations. Nevertheless, when LysoSensor^? was compared with SYBR‐14/PI, the agreement was high (mean of differences: ?0.05%; coefficient of agreement: 5.07%). Repeatability of both methods was high and similar. LysoSensor^? did not seem to specifically stain the acrosome, but it may accumulate in the cytoplasm and label viable spermatozoa. Therefore, LysoSensor^? might not be used as an acrosomal probe in ram spermatozoa, but it could be used in other kind of studies, taking advantage of its pH sensitivity.     

Interactions between different media and follicle‐stimulating hormone supplementation on in vitro culture of preantral follicles enclosed in ovarian tissue derived from collared peccaries (Pecari tajacu Linneaus, 1758)

The aim was to verify the effect of follicle‐stimulating hormone (FSH) supplementation to α‐MEM+ or TCM199+ media on the in vitro development of ovarian preantral follicles (PFs) derived from collared peccaries. Ovaries (n = 5 pairs) were collected and divided into fragments destined to control group (non‐cultured) or treatments that were cultured for 7 days. The PFs morphology, growth and activation were evaluated by classical histology. The immunohistochemistry markers Ag‐NOR and PCNA were used for nuclear proliferation analysis, and the picrosirius red labelling was used for ovarian extracellular matrix (ECM) evaluation. After 7‐day culture, only the TCM199+ treatment maintained the proportion of intact PFs similar to day 1 (63.2%), but no differences were found among treatments (p > .05). In addition, a significant increase in the growing follicles proportion was verified for all the treatments, indicating follicular activation (p > .05). By the Ag‐NOR analysis, only the TCM199+/FSH maintained the nuclear proliferation similar to the first day (p > .05). The picrosirius red staining revealed that the ECM remained intact in all the treatments (p > .05). We suggest the use of TCM199+ medium supplemented of FSH for the in vitro development of peccaries PFs under 7‐day culturing conditions.     

Intranasal Osteosarcoma in a Dog—A Case Report

Osteosarcoma is the most commonly reported bone tumor in dogs, typically affecting the axial and appendicular skeleton. Involvement of the paranasal sinuses and the nasal cavity is extremely rare. This report describes a 6-year-old, female, cross-breed dog presented with signs of 90 days duration and diagnosed with a mass located in the right nasal cavity. Radiography, rhinoscopy, and computed tomography findings suggested the presence of an intranasal tumor. Osteosarcoma in the nasal cavity was diagnosed based on histopathological examination and clinical and imaging findings. Survival time was 12 months from diagnosis.     

Experimental acute canine monocytic ehrlichiosis: clinicopathological and immunopathological findings

Clinical signs, humoral and cellular immune responses, and microscopic and gross tissue alterations resulting from acute experimental Ehrlichia canis infection in dogs were studied. Four dogs were inoculated with E. canis and four were used as uninfected controls. After a 10-14-day incubation period, infected dogs developed pyrexia up to 41 degrees C for 6-8 days. Antibody titers to E. canis antigen were demonstrable in all inoculated dogs at 30 days post-infection. Necropsy of infected animals revealed pale mucous membranes, generalized lymphadenopathy, splenomegaly, edema and ascites. Microcopically, the main lesions were: lymphoreticular hyperplasia in cortical areas of lymph nodes and spleenic white pulp, periportal accumulation of mononuclear cells and centrolobular fatty degeneration of the liver. Kidneys presented with glomerulonephritis characterized by interstitial mononuclear infiltration. Immunophenotyping of lymphocytes from lymph nodes and spleen sections displayed alterations in IgG, IgM, CD3+ and CD8+ cells population in infected dogs.     

Evaluation of the infectivity, length of infection, and immune response of a low-pathogenicity H7N2 avian influenza virus in specific-pathogen-free chickens

The H7N2 subtype of avian influenza virus (AIV) field isolate (H7N2/chicken/PA/3779-2/97), which caused the 1997-98 AIV outbreak in Pennsylvania, was evaluated for its infectivity, length of infection, and immune response in specific-pathogen-free (SPF) chickens. The composite findings of three clinical trials with various concentrations of virus indicated that this H7N2 subtype contained minimal pathogenicity for chickens. The concentration of the virus in the inoculum proved critical in the establishment of a productive infection in a chicken. Seven-day-old SPF chickens were not infected when inoculated with 10(0.7-2.0) mean embryo lethal dose (ELD50) of the H7N2 virus per bird. At this dose level, the immune response to this virus was not detected by the hemagglutination-inhibition (HI) test. Nonetheless, chickens at ages of 5 and 23 wk old tested were successfully infected when exposed to 10(4.7-5.7) ELD50 of H7N2 infectious doses per bird by various routes of administration and also by direct contact. Infected birds started shedding virus as early as 2 days postinoculation, and the period of virus shedding occurred mostly within 1 or 2 wk postinoculation (WPI). This H7N2 subtype of AIV induced a measurable immune response in all birds within 2 wk after virus exposure. Antibody titers were associated with AIV infectious doses and age of exposure of birds. Challenge of these infected birds with the same H7N2 virus at 5 and 10 WPI indicated the infective virus was recoverable from cloacal swabs at 3 days postchallenge and disappeared thereafter. In these challenged birds, the antibody levels as measured by the HI test spiked within 1-2 wk.     

First molecular detection of canine herpesvirus 1 (CaHV-1) in the Eastern Brazilian Amazon

BackgroundCanine herpesvirus type 1 (CaHV-1) infects dogs and is associated with neonatal deaths and reproductive, ocular, neurological, and respiratory problems. In Brazil, reports of CaHV-1 have been restricted to the southeast and south regions, particularly in municipalities in the state of Rio Grande do Sul.ObjectivesTo assess the presence and variability of CaHV-1 in canine populations in the state of Pará, North Brazil.MethodsBiological samples from 159 dogs from 4 municipalities in the State of Pará were evaluated using polymerase chain reaction and phylogenetic analyses, with the target being the viral enzyme, thymidine kinase.ResultsCaHV-1 was detected in 13 dogs (8.2%), with 2 animals being from the municipality of Santa Bárbara do Pará, 8 from Algodoal Island, 2 from Salinópolis, and one from Capanema. The study sequences revealed 100% identity among themselves and 64% to 100% identity with the other nucleotide sequences from Australia, Brazil, United Kingdom, and United States, including 100% identity with the 2002 isolate from Australia. The 1996 isolate from France was grouped in a branch that was different from the sequence of this study.ConclusionsThis study presents the first molecular detection of CaHV-1 in dogs from the Amazon region in northern Brazil. The nucleotide identity between the strains and cytosine insertion in the sequences isolated in this study suggests at least 2 strains of CaHV-1 circulating in Brazil (Pará and BTU-1).     

The use of porcine small-intestinal submucosa for abdominal wall reconstruction – a clinical case

A seven-year-old miniature dachshund was presented with a large contaminated bite wound centred on the left costal arch. Survey radiographs revealed a pneumomediastinum, a mid-body fracture of the left 11th rib and luxation of the xiphisternum. The compromised skin, ribs, xiphisternum, diaphragm and abdominal wall were surgically debrided. This resulted in a large combined thoracic and abdominal wall defect. The thoracic cavity was closed by diaphragmatic advancement. The abdominal wall defect was partially closed by advancing local abdominal musculature. A cranial abdominal mid-line defect remained and was repaired using two sheets of lyophilised porcine small-intestinal submucosa. Dehiscence of the skin exposing the SIS graft was seen but the abdominal repair remained intact. Eighteen months after the initial injury, the dog remained clinically well and no associated complications were reported.     

Dynamics and efficacy of sulfentrazone,flumioxazin, and isoxaflutole herbicides applied on eucalyptus harvest residues

New Forests - Eucalyptus harvests generate residues that remain on the soil and affect the efficacy of preemergent herbicides in the following planting cycle. The objective of this work was to...     

Post-fire soil respiration in relation to burnt wood management in a Mediterranean mountain ecosystem

After a wildfire, the management of burnt wood may determine microclimatic conditions and microbiological activity with the potential to affect soil respiration. To experimentally analyze the effect on soil respiration, we manipulated a recently burned pine forest in a Mediterranean mountain (Sierra Nevada National and Natural Park, SE Spain). Three representative treatments of post-fire burnt wood management were established at two elevations: (1) “salvage logging” (SL), where all trees were cut, trunks removed, and branches chipped; (2) “non-intervention” (NI), leaving all burnt trees standing; and (3) “cut plus lopping” (CL), a treatment where burnt trees were felled, with the main branches lopped off, but left in situ partially covering the ground surface. Seasonal measurements were carried out over the course of two years. In addition, we performed continuous diurnal campaigns and an irrigation experiment to ascertain the roles of soil temperature and moisture in determining CO₂ fluxes across treatments. Soil CO₂ fluxes were highest in CL (average of 3.34 ± 0.19 μmol m⁻² s⁻¹) and the lowest in SL (2.21 ± 0.11 μmol m⁻² s⁻¹). Across seasons, basal values were registered during summer (average of 1.46 ± 0.04 μmol m⁻² s⁻¹), but increased during the humid seasons (up to 10.07 ± 1.08 μmol m⁻² s⁻¹ in spring in CL). Seasonal and treatment patterns were consistent at the two elevations (1477 and 2317 m a.s.l.), although respiration was half as high at the higher altitude.Respiration was mainly controlled by soil moisture. Watering during the summer drought boosted CO₂ effluxes (up to 37 ± 6 μmol m⁻² s⁻¹ just after water addition), which then decreased to basal values as the soil dried. About 64% of CO₂ emissions during the first 24 h could be attributed to the degasification of soil pores, with the rest likely related to biological processes. The patterns of CO₂ effluxes under experimental watering were similar to the seasonal tendencies, with the highest pulse in CL. Temperature, however, had a weak effect on soil respiration, with Q₁₀ values of ca. 1 across seasons and soil moisture conditions. These results represent a first step towards illustrating the effects of post-fire burnt wood management on soil respiration, and eventually carbon sequestration.