The Effect of Season on the Histologic and Histomorphometric Appearance of the Equine Pituitary Gland

The objective of this study was to determine the effect of season on the histologic and histomorphometric appearance of the normal equine pituitary gland. Pituitary glands were collected at necropsy from 121 horses throughout the year. Plasma was also collected from 59 of these horses before euthanasia. Hematoxylin and eosin stained median sagittal sections of each pituitary were evaluated and histologically graded by three pathologists. Histomorphometric analysis was performed on the same slides. Plasma α-melanocyte stimulating hormone was measured by radioimmunoassay in a subset of horses (n = 59). A total of 118 pituitary glands were included in the study after exclusions were made on the basis of the presence of pars intermedia (PI) adenomas (>5 mm). There was a positive correlation between PI hormone concentration (α-melanocyte stimulating hormone) and PI area. Pituitary gland measurements and grades from samples collected in the fall were compared with those collected in the nonfall months using t-test. The PI area, total pituitary area, and PI/total pituitary ratio were significantly greater in the fall compared with nonfall months (P < .0001, P < .01, P < .0001, respectively). Pituitary grades were also higher in the fall compared with nonfall months (P < .001). There was no seasonal difference in pars distalis or pars nervosa area. The results of this study show that the normal equine pituitary shows seasonal changes in appearance and size. These changes must be considered when using postmortem histologic evaluations in the diagnosis of pituitary pars intermedia dysfunction or for validation of antemortem diagnostic tests.     

Digital hypothermia inhibits early lamellar inflammatory signalling in the oligofructose laminitis model

Reasons for performing study: The pathophysiological events inhibited by prophylactic digital hypothermia that result in reduction of the severity of acute laminitis are unknown. Objectives: To determine if digital hypothermia inhibits lamellar inflammatory signalling during development of oligofructose (OF) induced laminitis. Methods: Fourteen Standardbred horses were given 10 g/kg bwt OF by nasogastric tube with one forelimb (CRYO) continuously cooled by immersion in ice and water and one forelimb (NON‐RX) at ambient temperature. Lamellae were harvested prior to the onset of lameness (24 h post OF administration, DEV group, n = 7) or at the onset of lameness (OG1 group, n = 7). Lamellar mRNA was purified and cDNA produced for real time‐quantitative PCR analysis of mRNA concentrations of cytokines (IL‐6, IL‐1β, IL‐10), chemokines (CXCL1, CXCL6, CXCL8/IL‐8, MCP‐1, MCP‐2), cell adhesion molecules (ICAM‐1, E‐selectin), COX‐2 and 3 housekeeping genes. Data were analysed (NON‐RX vs. CRYO, NON‐RX vs. archived control [CON, n = 7] lamellar tissue) using nonparametric tests. Results: Compared with CON, the OG1 NON‐RX had increased (P<0.05) lamellar mRNA concentrations of all measured mediators except IL‐10, IL‐1β and MCP‐1/2, whereas only CXCL8 was increased (P<0.05) in DEV NON‐RX. Within the OG1 group, CRYO limbs (compared with NON‐RX) had decreased (P<0.05) mRNA concentrations of the majority of measured inflammatory mediators (no change in MCP‐1 and IL‐10). Within the DEV group, mRNA concentrations of CXCL‐1, ICAM‐1, IL‐1β, CXCL8 and MCP‐2 were decreased (P<0.05) and the anti‐inflammatory cytokine IL‐10 was increased (compared with NON‐RX limbs; P<0.05). Conclusions: Digital hypothermia effectively blocked early lamellar inflammatory events likely to play an important role in lamellar injury including the expression of chemokines, proinflammatory cytokines, COX‐2 and endothelial adhesion molecules. Potential relevance: This study demonstrates a potential mechanism by which hypothermia reduces the severity of acute laminitis, and may help identify molecular targets for future laminitis intervention.     

Hindlimb laminar inflammatory response is similar to that present in forelimbs after carbohydrate overload in horses

Reasons for performing study: A significant proinflammatory response is known to occur in the forelimb lamina after carbohydrate administration. As the hindlimbs are often less affected by laminitis compared with the forelimbs, we assessed hindlimb inflammatory response in the early stages of carbohydrate‐induced laminitis to determine whether differences in the response existed. Objective: To determine whether a similar proinflammatory response occurs in the hindlimb laminae to that previously reported for the forelimb. Methods: Archived laminar samples from 12 horses administered 17.6 g of starch (85% corn starch, 15% wood flour)/kg bwt via nasogastric tube that were anaesthetised either after developing a temperature >38.9°C (DEV; n = 6) or at the onset of Obel grade 1 lameness (OG1; n = 6) were used in addition to 6 control horses (CON) that were anaesthetised 24 h after administration of water. Real‐time quantitative polymerase chain reaction for selected proinflammatory mediators and MAC387 immunohistochemistry were performed. The data were analysed nonparametrically to compare groups. Results: Increases in laminar MAC387‐positive leucocytes and laminar messenger ribonucleic acid (mRNA) concentrations (P<0.05) for interleukin‐1β, interleukin‐6, cyclo‐oxygenase‐2, chemokine (C‐X‐C motif)ligand (CXCL)1 and CXCL8 were present in both fore‐ and hindlimb laminae from horses with OG1 lameness. Both CXCL1 and CXCL8 were also increased in forelimb and hindlimb laminae in the DEV horses. Conclusions: Administration of carbohydrate resulted in a similar inflammatory response in the hindlimb laminae to that previously reported for the forelimb laminae. These findings suggest that other factors, such as weightbearing, may play an important role in the development of laminitis after a systemic inflammatory condition develops. Potential relevance: Evidence of inflammation in the hindlimb laminae suggests that the hindfeet should be addressed in the septic horse at risk for laminitis; however, laminitis is often less severe in the hindlimbs due to other factors, such as weightbearing and hoof angle.     

Electroretinogram responses of the normal thoroughbred horse sedated with detomidine hydrochloride

Purpose The main objective was to record electroretinogram (ERG) parameters of normal thoroughbred mares using the HMsERG, a mini-Ganzfeld electroretinographic unit, and a contact lens electrode. The second objective was to determine whether IV detomidine hydrochloride at 0.015?mg/kg is consistently an effective choice for sedation of horses undergoing this ERG protocol. Methods The study population consisted of 30 normal thoroughbred mares. ERG data were harvested using a protocol that included three different light intensities (10, 3000, and 10?000?mcd?s/m(2) ) and a 30-Hz flicker at 3000?mcd?s/m(2) . Results Mean, median, standard deviation, and estimated normal ranges using the 5-95% of the data for a- and b-wave implicit times (IT), amplitudes (AMP), and b/a ratios were reported. Scotopic results at low intensity (10?mcd?s/m(2) ) had estimated ranges for b-wave IT of 41.8-72.9?ms and AMP of 19.8-173.3?μV. Middle intensity (3000?mcd?s/m(2) ) a-wave IT was 13.2-14.7?ms with a-wave AMP of 68.4-144?μV; the b-wave IT was 28.7-41.5?ms with b-wave AMP of 105.7-271.5?μV; and the b/a ratio was 0.95-2.71. The high-intensity (10?000?mcd?s/m(2) ) average recordings showed an a-wave IT of 13-14.9?ms, a-wave AMP of 85.7-186.8?μV; b-wave IT of 26.6-45.4?ms, b-wave AMP of 104.7-250.6?μV; and a b/a wave ratio of 0.7-2.0. The 30-Hz cone flicker showed an IT of 22.8-28.9?ms and AMP of 44.1-117.1?μV. Conclusions Results of normal thoroughbred ERG responses are reported. The protocol proved to be simple and safe and provided consistent results.     

The prevalence and clinical relevance of hyperkalaemia in calves with neonatal diarrhoea

One hundred and twenty-four calves with neonatal diarrhoea were investigated in order to assess the prevalence of hyperkalaemia and the associated clinical signs. Hyperkalaemia (potassium concentration >5.8 mmol/L) was recognized in 42 (34%) calves and was more closely associated with dehydration than with decreases in base excess or venous blood pH. In 75 calves with normal blood concentrations of D-lactate (i.e. ?3.96 mmol/L), K concentrations were moderately correlated with base excess values (r = ?0.48, P < 0.001). In contrast, no significant correlation was observed in 49 calves with elevated D-lactate. Only three hyperkalaemic calves had bradycardia and a weak positive correlation was found between heart rate and K concentrations (r = 0.22, P = 0.014). Ten of the 124 calves had cardiac arrhythmia and of these seven had hyperkalaemia indicating that cardiac arrhythmia had a low sensitivity (17%) but a high specificity (96%) as a predictor of hyperkalaemia.In a subset of 34 calves with base excess values ??5 mmol/L and D-lactate concentrations <5 mmol/L (of which 22 had hyperkalaemia), changes in posture/ability to stand could be mainly explained by elevations of K concentrations (P < 0.001) and to a lesser extent by increases in L-lactate concentrations (P = 0.024). Skeletal muscle weakness due to hyperkalaemia alongside hypovolaemia may produce a clinical picture that is similar to that in calves with marked D-lactic acidosis. However, since reductions in the strength of the palpebral reflex are closely correlated with D-lactate concentrations, a prompt palpebral reflex can assist the clinical prediction of hyperkalaemia in calves presenting with a distinct impairment in their ability to stand (specificity 99%, sensitivity 29%).     

Outbreak of Philophthalmus gralli in four greater rheas (Rhea americana)

Using slit‐lamp biomicroscopy, conjunctival biopsy, and morphological identification, a flock of four Greater rheas (Rhea americana) in Arizona were diagnosed with conjunctivitis secondary to Philophthalmus gralli (P. gralli) infection. Aquatic snails from the exhibit’s water source were identified as Melanoides tuberculatus, a known vector for P. gralli. Comparison of partial sequences of DNA regions from P. gralli adults removed from the rheas and metacercariae from the aquatic snails demonstrated a 100% match, confirming the source of infection. The flock was divided into two treatment groups: the most severely affected rheas received both manual removal of trematodes and praziquantel 1% ointment OU q12 h and the least severely affected rheas were only given praziquantel 1% ointment OU q12 h. The rheas were permanently relocated away from the infected water source and aquatic snails. Initial resolution was seen at 17 weeks in the most severely affected rhea, which had 675 adult P. gralli removed and topical praziquantel. The two rheas that only received topical praziquantel showed resolution within 3 and 15 weeks. Current recommendations for treating P. gralli include: manual removal of trematodes, topical praziquantel 1% ointment, and relocation away from infected water sources and aquatic snails.     

Source relationships between streambank soils and streambed sediments in a mercury-contaminated stream

Purpose

In contaminated streams, understanding the role of streambank and streambed source contributions is essential to developing robust remedial solutions. However, identifying relationships can be difficult because of the lack of identifying signatures in source and receptor pools. East Fork Poplar Creek (EFPC) in Oak Ridge, TN, USA received historical industrial releases of mercury that contaminated streambank soils and sediments. Here, we determined relationships between the contaminated streambank soils and sand-sized streambed sediments.

Materials and methods

Field surveys revealed the spatial trends of the concentrations of inorganic total mercury (Hg) and methyl mercury (MeHg), Hg lability as inferred by sequential extraction, particle size distribution, and total organic carbon. Statistical tests were applied to determine relationships between streambank soil and streambed sediment properties.

Results and discussion

Concentrations of Hg in streambank soils in the upper reaches averaged 206 mg kg^?1 (all as dry weight) (n?=?457), and 13 mg kg^?1 in lower reaches (n?=?321), while sand-sized streambed sediments were approximately 16 mg kg^?1 (n?=?57). Two areas of much higher Hg and MeHg concentrations in streambank soils were identified and related to localized higher Hg concentrations in the streambed sediments; however, most of the streambank soils have similar Hg concentrations to the streambed sediments. The molar ratio of Hg to organic carbon, correlation between MeHg and Hg, and particle size distributions suggested similarity between the streambank soils and the fine sand-sized fraction (125–250 μm) collected from the streambed sediments. Mercury in the fine sand-sized streambed sediments, however, was more labile than Hg in the streambank soils, suggesting an in-stream environment that altered the geochemistry of sediment-bound Hg.

Conclusions

This study revealed major source areas of Hg in streambank soils, identified possible depositional locations in streambed sediments, and highlighted potential differences in the stability of Hg bound to streambank soils and sediments. This work will guide future remedial decision making in EFPC and will aid other researchers in identifying source–sink linkages in contaminated fluvial systems.

     

Wetting of agricultural soil measured by a simplified capillary rise technique

We describe the use of a capillary rise method to measure the wettability of 10 samples of agricultural soil from Rothamsted long-term experimental sites. The samples have very similar clay contents, but organic carbon (C) contents range from 11.5 to 31.2 g kg⁻¹. Their wetting rates were interpreted by an improved method of data analysis, consistent with the Washburn equation, and showed an increase in the effective contact angle between the water meniscus and the soil with increasing C content. This corresponds to a decrease in wettability with increasing C content, and accords with other results reported in the literature. By contrast with water, we found that capillary rise for n -hexane into soil did not depend on the soil's bulk density or C content. A priori calculations of the expected wetting rates from fluid properties and an effective hydraulic radius estimated by other methods gave magnitudes and trends that agreed with our experimental data. The results show that estimates of effective hydraulic radius can provide a useful approximation for characterizing soil wetting, but that further modelling should be carried out.     

Soil and Atmospheric Inputs to PAH Concentrations in Salt Marsh Plants

Polycyclic aromatic hydrocarbons (PAHs) were measured in two wetland plant species grown outdoors in pots of sediment contaminated with up to 730,000 μg/kg total PAHs. After approximately 3 months, the plants were harvested, cleaned, and analyzed for an expanded suite of PAHs which included both the 16 priority PAHs and 22 alkyl PAH homologs. Sediment and air samples were also collected and analyzed. PAH compounds were detected in all of the samples, with the highest concentrations in the contaminated sediment. The root sample concentrations were generally about one order of magnitude lower than that of the sediment, and were strongly correlated with the concentration in the sediment in which they were grown. The concentrations in foliage were much lower and did not correlate with sediment concentration. Concentrations of low molecular weight PAH compounds detected in the foliage were not significantly lower in plants grown in control sediments, suggesting that the sediment is not the primary source of these PAHs. Several high molecular weight PAHs were detected only in plants grown in contaminated sediment. Plants of both species grown in control sediment were larger than plants grown in contaminated sediment.