Cytological analysis of bronchoalveolar lavage fluid acquired by bronchoscopy in healthy ferrets: A pilot study

The objective of this study was to investigate the normal cytological evaluation of bronchoalveolar lavage (BAL) fluid in healthy adult ferrets (N = 12). These ferrets underwent bronchoscopy and BAL using sterile saline [1.5 mL/kg body weight (BW)]. Percentage of fluid recovered, total leukocyte count, differential leukocyte count, and cell count of the epithelial lining fluid (ELF) were determined. The mean percentage of lavage volume recovered from the right lung and left lung were 67.8 ± 14.9% and 69.7 ± 20.0%, respectively. Gender (P = 0.12) and weight (P = 0.17) did not significantly affect the mean percentage of recovered volume. The mean percentage of recovered volume (P = 0.47) and the mean leukocyte count (P = 0.17) from the right and left lung were not significantly different. Macrophages were the main leukocyte component of the lavages, followed by neutrophils, lymphocytes, and eosinophils. The mean proportion of ELF in BAL fluid was 9.3 ± 3.7% v/v. Bronchoscopy is clinically useful for collecting good quality BAL samples for cytological analysis in ferrets. The leucocyte differential was established, which may help veterinarians to make better clinical decisions when treating respiratory disease. Further studies are required with a larger group in order to establish the healthy reference intervals for BAL values in ferrets.     

Cardiovascular images: vascular hamartoma of the mitral valve in a horse

An 8-month-old Hanoverian gelding was presented with a history of cardiac murmurs that were not apparent as a foal nor reported at the time of castration. Major echocardiographic findings included mitral valvular thickening, functional stenosis, and mitral regurgitation of sufficient severity to cause diastolic and systolic cardiac murmurs, left-sided volume overload, and pulmonary hypertension. Due to the hemodynamic severity of the lesion and poor prognosis for future performance and longevity, euthanasia was elected. On gross postmortem examination, there was focal fibrous epicarditis affecting the heart base, and the left atrium was moderately dilated. The mitral valve surface was irregular and contained several nodules along the atrial face of the cusp. Histologically, this lesion was diagnosed as a vascular hamartoma, which is rarely reported in veterinary species and has not been described in heart valves. This benign proliferative lesion, and concurrent valvular dysfunction, was associated with an unusual manifestation of clinically evident disease and should be differentiated from common incidental valvular lesions such as hematocysts.     

Changes in aggregate stability,nutrient status,indigenous microbial populations,and seedling emergence,following inoculation of soil withNostoc muscorum

The potential of the N₂-fixing cyanophyteNostoc muscorum for improving the aggregate stability of a poorly structured silt loam soil was studied in a greenhouse experiment. Inoculum rates were 1.61×10⁵ cells g^-1 soil dry weight (low rate) and 4.04×10⁵ cell g^-1 soil dry weight (high rate), approximately equivalent to a field application of 2 and 5 kg ha^-1 cells dry weight, respectively.N. muscorum numbers had increased 8-fold (low rate) and 10-fold (high rate) by 300 days after inoculation, indicating not only survival but proliferation. Increases in soil polysaccharides, determined as soil carbohydrate C, were 2.96–3.49 time the values in the non-inoculated soils and aggregate stability had incrased by an average of 18% on day 300. Inoculation withN. muscorum also had a pronounced effect on soil chemical and biological properties, with total C increasing by 50–63% and total N increasing by 111–120%. Increases in the soil indigenous microbial population were recorded, with numbers of bacteria 500, fungi 16, and actinomycetes 48 times the non-inoculated values on day 300 in the high-rate soil. The emergence of lettuce seedlings (Lactuca sativa var. Saladin) in undisturbed inoculated 300-day soils was 56% (low rate) and 52% (high rate) higher than in non-inoculated soils. However, homogenising soils and irrigating (to smulate ploughing and surface crusting) significantly reduced this increase in both treatments, although emergence in inoculated soils was still greater by 45% (low) and 24% (high). It is recommended that inoculated soils be left undisturbed prior to planting. The effects ofN. muscorum on soil physical, chemical, and biological properties indicate the possible benefits of cyanobacteria as soil inoculants, not only for the improvement of soil aggregate stability but also as a means of improving seedling emergence.     

Nitrogen immobilization by wood-chip application: Protecting water quality in a northern hardwood forest

Forest harvesting disrupts the nitrogen cycle, which may affect stream water quality by increasing nitrate concentrations, reducing pH and acid neutralizing capacity, and mobilizing aluminum and base cations. We tested the application of wood chips derived from logging slash to increase immobilization of N after harvesting, which should reduce nitrate flux to streams. In August 2004, a stand of northern hardwoods was patch-clearcut in the Catskill Mountains, NY, and four replicates of three treatments were implemented in five 0.2-ha cut patches. Wood chips were applied to the soil surface at a rate equivalent to the amount of slash smaller than eight inches in diameter (1× treatment). A second treatment doubled that rate (2×), and a third treatment received no chips (0×). Additionally, three uncut reference plots were established in nearby forested areas. Ion exchange resin bags and soil KCl-extractions were used to monitor nitrate availability in the upper 5–10 cm of soil approximately every seven weeks, except in winter. Resin bags indicated that the wood chips retained 30% or 42% of the nitrate pulse, while for KCl extracts, the retention rate was 78% or 100% of the difference between 0× and uncut plots. During the fall following harvest, wood-chip treated plots had resin bag soil nitrate concentrations about 25% of those in 0× plots (p = 0.0001). In the first growing season after the cut, nitrate concentrations in wood-chip treated plots for KCl extracts were 13% of those in 0× treatments (p = 0.03) in May and about half those in 0× treatments (p = 0.01) in July for resin bags. During spring snowmelt, however, nitrate concentrations were high and indistinguishable among treatments, including the uncut reference plots for resin bags and below detection limit for KCl extracts. Wood chips incubated in litterbags had an initial C:N of 125:1, which then decreased to 70:1 after one year of field incubation. These changes in C:N values indicate that the wood-chip application can potentially immobilize between 19 and 38 kg N ha⁻¹ in the first year after harvesting, depending on the rate of wood-chip application. Our results suggest that the application of wood chips following harvesting operations can contribute to the protection of water quality and warrant additional research as a new Best Management Practice following cutting in regions that receive elevated levels of atmospheric N deposition.     

Effect of body condition score and reuse of progesterone‐releasing intravaginal devices on conception rate following timed artificial insemination in Nelore cows

This study had the aim of investigating the efficiency of timed artificial insemination (TAI) through the progesterone‐releasing intravaginal device (PRID), used in new condition and for the second and third times in Nelore cows. The effects of device reuse and body condition score (BCS) on the conception rate (CR) were evaluated in 1,122 multiparous Nelore cows (mean BCS of 2.7 ± 0.4), which were randomly distributed into three groups that received new (n = 330), once (n = 439) and twice used (n = 353) PRID. Among the 1,122 females that underwent TAI, 573 became pregnant, thus representing an overall CR of 51.06%. Cows with BCS between 2.75 and 4.0 had greater (p < .0001) CR (69.75%) than cows with BSC of 2.0–2.5 (32.98%). It was observed that the CR through using PRID was 60.00%, 51.71% and 41.93% for new, once and twice used PRID, respectively, with difference between all groups (p < .0001). Under tropical conditions, animals with BCS greater than 2.5 had a higher CR, and the CR decreased proportionally with the number of times that the PRID had been used.     

Studies of the adsorption of paraquat on soluble humic fractions by gel filtration and ultrafiltration techniques

The adsorption of paraquat dichloride (1,1′-dimethyl-4,4′-bipyridylium dichloride) on a soluble sodium humate fraction of a Fenland soil was studied by gel filtration (on Sephadex G10 and G100) and by ultrafiltration (through an Amicon Diaflo UM-2 ultrafilter). Both methods depend upon the separation, on a molecular weight basis, of the unadsorbed molecules of herbicide from the adsorption complex (consisting of polymeric organic materials and the adsorbed paraquat). Separations were obtained on columns of Sephadex G10 (Method I) and in the ultrafiltration experiments (Method II), and isotherms were prepared from data for adsorption in water (by Method II) and in sodium chloride (by Methods I and II) solutions. Results from the two methods were comparable over the concentration range examined. The increased adsorption of paraquat by Na⁺-compared with Ca²⁺-humate is explained on the basis of the selectivity sequence of humate for exchangeable cations. Attempts to prepare isotherms from gel filtration data, for the adsorption of paraquat on two soluble model humic polymers (polyacrylic acid and a polymer prepared by the oxidative coupling of benzoquinone and ammonium chloride) were unsuccessful because binding to the gel matrix did not permit quantitative recoveries of the adsorption complexes. Paraquat was adsorbed to the same extent on each of four fractions of Na⁺-humate separated on Sephadex G100.     

Isolation and Culture of Ovine and Bubaline Small and Large Pre-antral Follicles: Effect of Cyclicity and Presence of a Dominant Follicle

Studies were conducted to examine the effects of the cyclicity and the presence of a dominant follicle (DF) in ovary on the recovery and in vitro growth of pre-antral follicles (PFs) in sheep and buffalo. Small pre-antral follicles (SPFs, 100–250 μm) and large pre-antral follicles (LPFs, 250–450 μm) were isolated from slaughterhouse ovaries in the breeding seasons by a mechanical and enzymatic method. The sheep and buffalo PFs were cultured in vitro for 6 and 15 days, respectively, and examined for their growth, survival and antrum formation rates and growth rates of oocytes in cultured pre-antral follicles. The follicles of the sheep and buffalo were recovered and cultured simultaneously within replicates. The recovery rates (number per ovary) of both SPFs and LPFs were significantly (p < 0.05) higher in cyclic ewes (SPFs: 22.0 ± 3.3 vs 12.1 ± 2.6 and LPFs: 16.0 ± 3.6 vs 9.2 ± 1.8) and buffaloes (SPFs: 9.2 ± 1.3 vs 4.1 ± 1.0 and LPFs: 10.3 ± 2.7 vs 5.4 ± 0.7) compared with those recovered from acyclic ones. Presence of a DF in ovary significantly (p < 0.05) reduced the recovery rates of LPFs in ewes (9.06 ± 2.7 vs 16.4 ± 3.8) but had no effect in buffalo. Cyclicity of animals or follicular dominance had no effects on in vitro growth, survival and antrum formation rates and growth rates of oocytes in cultured PFs of SPFs and LPFs in both sheep and buffalo. The in vitro growth, survival and antrum formation rates of LPFs and growth rates of oocytes in cultured LPFs were significantly (p < 0.05) higher than those observed in SPFs in both sheep and buffalo. The overall recovery and growth rates of the PFs were lower in buffaloes compared with ewes.     

Purification of secretory immunoglobulin A from milk of the brushtail possum (Trichosurus vulpecula)

AIM: To identify and purify secretory immunoglobulin A (sIgA), a key effecter molecule in mucosal immune responses, from milk of the brushtail possum (Trichosurus vulpecula).

METHODS: Milk samples were collected from female possums with pouch young, and clarified by centrifugation and precipitation methods. The clarified fraction was purified by gel filtration and affinity chromatography to yield sIgA. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting techniques were used to assess the purity of the final product, and to identify the heavy (H) chain, light (L) chain and secretory component (SC) of possum sIgA.

RESULTS: Immunoblotting, using antibodies raised against cloned possum sIgA SC and H-chain, and a synthetic peptide fragment of the H-chain, confirmed the identity of the purified protein. The N-terminal amino acid sequence of purified possum sIgA showed strong homology to reported sequences of H-chain variable regions of marsupial immunoglobulins.

CONCLUSIONS: Milk was shown to be a convenient source of mucosal secretion containing sIgA, and a process involving 2 precipitation and 2 chromatography steps produced purified sIgA. This IgA preparation will prove useful for the generation of sIgA-specific immunological reagents for measurement of immune responses in the development of mucosal-based vaccines for biological control of possums.