Apoptosis in the Antral Follicles of Swamp Buffalo and Cattle Ovary: TUNEL and Caspase-3 Histochemistry

The present study was carried out to investigate the pattern of apoptosis in the healthy antral and atretic follicles of Philippine swamp buffaloes (BU) in comparison with Holstein-Friesian (HF) cows. Paraffin sections of healthy follicles and various stages of atretic follicles were stained using the terminal deoxynucleotidyl transferase (TdT)-mediated biotinylated deoxyuridine triphosphates (dUTP) nick end-labelling (TUNEL) method to detect DNA fragmentation and cleaved caspase-3 antibody to detect cells committed to undergo apoptosis. Five equidistant areas of a follicle were counted for the presence of TUNEL- and caspase-3-positive cells. Healthy follicles of BU and HF contained no TUNEL-positive cells in the granulosa and theca layer but showed some caspase-3 positivity. The granulosa layer of advanced atretic follicles showed a significantly higher frequency of caspase-3 positivity than the healthy and early atretic follicles in both breeds. The frequency of caspase-3-positive cells of BU was significantly higher than HF in the granulosa layer of healthy, early atretic and advanced atretic follicles. In the theca interna layer, BU and HF showed a significantly lower and higher frequency of TUNEL-positive cells in the late atretic follicles compared with advanced atretic follicle, respectively. However, the frequency of caspase-3-positive cells of both BU and HF in the late atretic follicles was significantly higher than the advanced atretic follicles in the theca interna layer. These results indicate that caspase-3 and DNA fragmentation is involved in the buffalo ovarian apoptotic process.     

Cell Proliferation in the Atretic Follicles of Buffalo and Cattle Ovary

The present study was carried out to describe the proliferative activity of granulosa and theca cells in healthy antral and atretic follicles of Philippine buffaloes (BU) and Holstein-Friesian (HF) cows. Paraffin sections of ovary were immunostained with mouse monoclonal antibody to proliferating cell nuclear antigen (PCNA). Then the follicles were classified into healthy and various stages of atretic follicles. The granulosa layer of healthy follicles had a significantly higher frequency of PCNA-positive cells than the early and advanced atretic follicles in both breeds. In the theca interna, significantly reduced populations of the PCNA-positive cells were found in both breeds as atresia progressed. Moreover, HF had significantly higher PCNA-positive cells in the theca interna of healthy, early atretic and advanced atretic follicles than BU. A reduction of PCNA-positive cells during atresia was also noted in the theca externa in both animals although differences were not significant. The results of the present work suggest that the proliferative activity of granulosa and theca cells decreases in association with follicular atresia in the BU similar to HF. Furthermore, a significantly deficient cell proliferative activity of theca interna was found in BU compared with HF.     

Ergovaline does not alter the severity of ryegrass staggers induced by lolitrem B

AIM: To investigate a possible interaction between lolitrem B and ergovaline by comparing the incidence and severity of ryegrass staggers in sheep grazing ryegrass (Lolium perenne) containing lolitrem B or ryegrass containing both lolitrem B and ergovaline.

METHODS: Ninety lambs, aged approximately 6 months, were grazed on plots of perennial ryegrass infected with either AR98 endophyte (containing lolitrem B), standard endophyte (containing lolitrem B and ergovaline) or no endophyte, for up to 42 days from 2 February 2010. Ten lambs were grazed on three replicate plots per cultivar. Herbage samples were collected for alkaloid analysis and lambs were scored for ryegrass staggers (scores from 0–5) weekly during the study. Any animal which was scored ≥4 was removed from the study.

RESULTS: Concentrations of lolitrem B did not differ between AR98 and standard endophyte-infected pastures during the study period (p=0.26), and ergovaline was present only in standard endophyte pastures. Ryegrass staggers was observed in sheep grazing both the AR98 and standard endophyte plots, with median scores increasing in the third week of the study. Prior to the end of the 42-day grazing period, 22 and 17 animals were removed from the standard endophyte and AR98 plots, respectively, because their staggers scores were ≥4. The cumulative probability of lambs having scores ≥4 did not differ between animals grazing the two pasture types (p=0.41).

CONCLUSIONS AND CLINICAL RELEVANCE: There was no evidence for ergovaline increasing the severity of ryegrass staggers induced by lolitrem B. In situations where the severity of ryegrass staggers appears to be greater than that predicted on the basis of concentrations of lolitrem B, the presence of other tremorgenic alkaloids should be investigated.     

Microhabitat use of the diamond darter

The only known extant population of the diamond darter (Crystallaria cincotta) exists in the lower 37 km of Elk River, WV, USA. Our understanding of diamond darter habitat use was previously limited, because few individuals have been observed during sampling with conventional gears. We quantified microhabitat use of diamond darters based on measurements of water depth, water velocity and per cent substrate composition. Using spotlights at night‐time, we sampled 16 sites within the lower 133 km of Elk River and observed a total of 82 diamond darters at 10 of 11 sampling sites within the lower 37 km. Glides, located immediately upstream of riffles, were the primary habitats sampled for diamond darters, which included relatively shallow depths (<1 m), moderate‐to‐low water velocities (often < 0.5 m·s^?1) and a smooth water surface. Microhabitat use (mean ± SE) of diamond darters was estimated for depth (0.47 ± 0.02 m), average velocity (0.27 ± 0.01 m·s^?1) and bottom velocity (0.15 ± 0.01 m·s^?1). Substrate used (mean ± SE) by diamond darters was predominantly sand intermixed with lesser amounts of gravel and cobble: % sand (52.1 ± 1.6), % small gravel (12.2 ± 0.78), % large gravel (14.2 ± 0.83), % cobble (19.8 ± 0.96) and % boulder (1.6 ± 0.36). Based on our microhabitat use data, conservation and management efforts for this species should consider preserving glide habitats within Elk River. Spotlighting, a successful sampling method for diamond darters, should be considered for study designs of population estimation and long‐term monitoring.     

The WC1(+) gammadelta T-cell population in cattle: a possible role in resistance to intracellular infection

Intracellular infections are important in veterinary medicine and detailed understanding of the associated immune responses is needed for optimal development of strategies based on diagnosis and vaccination. It is generally accepted that cell-mediated immune responses are of greatest importance in intracellular infections and recent studies from several bovine models of infection indicate that WC1(+) gammadelta T-cells have a number of possible levels of involvement, which remain incompletely defined. Investigations of experimental infection with Mycobacterium bovis in cattle have indicated that WC1(+) gammadelta T-cells are among the first cells to accumulate at initial sites of infection, an observation which has been linked with decreased numbers of these cells in the circulation within days of infection. These WC1(+) gammadelta T-cells have been shown to respond in vitro, both to protein antigens and to non-protein, phosphate containing antigens of M. bovis and to be capable of producing IFN-gamma. Studies of M. bovis infection in calves depleted of WC1(+) gammadelta T-cells by monoclonal antibody have suggested that the presence of these cells is associated with development of a Th1-biased acquired immune response. In combination, these observations allow speculation regarding a possible role for WC1(+) gammadelta T-cells as a link between the innate and acquired immune systems which is instrumental in establishing an appropriate response.     

Dermoid sinus at the lumbosacral junction in an English springer spaniel

A dermoid sinus was identified in a springer spaniel that presented with hindlimb neurological deficits. The sinus was continuous with the dura mater at the level of the lumbosacral junction. The presence of hair and debris adjacent to neural tissue had elicited a myelitis. A dorsal laminectomy was required to allow complete surgical resection of the sinus. The dog's neurological status improved after treatment and this improvement was maintained over a five-month follow-up period.     

Preliminary clinical observations on the use of piroxicam in the management of rectal tubulopapillary polyps

Rectal tubulopapillary polyps were diagnosed in eight dogs following proctoscopy and mucosal pinch biopsy. Histological examination of the pinch biopsies revealed evidence of malignant transformation in three of the cases. The remaining cases were diagnosed as benign polyps. Inflammatory changes were observed in four cases. Seven dogs were treated with piroxicam suppositories and one with oral piroxicam. All dogs were re-examined after four to six weeks of piroxicam therapy and the extent of haematochezia, tenesmus and faecal mucus production was reduced in all cases. The owners of seven of the dogs considered the improvement in clinical signs to be good or excellent. Cases with and without evidence of inflammation responded equally well. This finding supports the hypothesis that piroxicam has an antineoplastic effect due to apoptosis and alteration in the cell cycle. Medical management with piroxicam may provide a non-invasive treatment option for dogs with rectal polyp formation in which surgical treatment is likely to be associated with complications such as incontinence, infection and wound breakdown, or where the owner declines such treatment.     

Relationship between physiological factors and clinical pain in dogs scored using a numerical rating scale

A study was designed to investigate the association between heart rate, respiratory rate and pupil dilation and a subjective pain score allocated using a numerical rating scale (NRS). Four groups of dogs (n = 17 to 20 per group) were included: orthopaedic surgery cases, soft tissue surgery cases, dogs with medical conditions and healthy dogs. Each dog was examined by five veterinary surgeons within a four-hour period. When examining each dog, the observers recorded heart rate, respiratory rate, pupil dilation (present or absent) and a pain score using a NRS (range 0 to 10). For surgical cases, all study assessments were carried out between 21 and 27 hours following the end of surgery. The correlation coefficients between physiological parameters and NRS score were small but, depending on the analysis, were significant for heart rate and NRS score (P < 0·05). However, the size of the coefficient, 0·168, indicated that the relationship was not biologically significant. There was no association between respiratory rate and NRS score. Analysis of the relationship between NRS score and pupil dilation indicated that there was an association between this factor and pain, depending on the analysis. This was significant for the surgical groups (P < 0·05) but not for the other groups.
These findings indicate that heart rate and respiratory rate are not useful indicators of pain in hospitalised dogs. It is also unlikely that pupil dilation will be a useful tool in the assessment of pain.     

Biodiversity management approaches for stream–riparian areas: Perspectives for Pacific Northwest headwater forests,microclimates, and amphibians

Stream–riparian areas represent a nexus of biodiversity, with disproportionate numbers of species tied to and interacting within this key habitat. New research in Pacific Northwest headwater forests, especially the characterization of microclimates and amphibian distributions, is expanding our perspective of riparian zones, and suggests the need for alternative designs to manage stream–riparian zones and their adjacent uplands. High biodiversity in riparian areas can be attributed to cool moist conditions, high productivity and complex habitat. All 47 northwestern amphibian species have stream–riparian associations, with a third being obligate forms to general stream–riparian areas, and a quarter with life histories reliant on headwater landscapes in particular. Recent recognition that stream-breeding amphibians can disperse hundreds of meters into uplands implies that connectivity among neighboring drainages may be important to their population structures and dynamics. Microclimate studies substantiate a “stream effect” of cool moist conditions permeating upslope into warmer, drier forests. We review forest management approaches relative to headwater riparian areas in the U.S. Pacific Northwest, and we propose scenarios designed to retain all habitats used by amphibians with complex life histories. These include a mix of riparian and upslope management approaches to address the breeding, foraging, overwintering, and dispersal functions of these animals. We speculate that the stream microclimate effect can partly counterbalance edge effects imposed by upslope forest disturbances, hence appropriately sized and managed riparian buffers can protect suitable microclimates at streams and within riparian forests. We propose one approach that focuses habitat conservation in headwater areas – where present management allows extensive logging – on sensitive target species, such as tailed frogs and torrent salamanders that often occur patchily. Assuming both high patchiness and some concordance among the distribution of sensitive species, protecting areas with higher abundances of these animals could justify less protection of currently unoccupied or low-density habitats, where more intensive forest management for timber production could occur. Also, we outline an approach that protects juxtaposed headwater patches, retaining connectivity among sub-drainages using a 6th-field watershed spatial scale for assuring well-distributed protected areas across forested landscapes. However, research is needed to test this approach and to determine whether it is sufficient to buffer downstream water quality and habitat from impacts of headwater management. Offering too-sparse protection everywhere is likely insufficient to conserve headwater habitats and biodiversity, while our alternative targeted protection of selected headwaters does not bind the entire forest landscape into a biodiversity reserve.