Comparative study of equine bone marrow and adipose tissue-derived mesenchymal stromal cells

Reasons for performing study: Mesenchymal stromal cells (MSCs) represent an attractive source for regenerative medicine. However, prior to their application, fundamental questions regarding molecular characterisation, growth and differentiation of MSCs must be resolved. Objectives: To compare and better understand the behaviour of equine MSCs obtained from bone marrow (BM) and adipose tissue (AT) in culture. Methods: Five horses were included in this study. Proliferation rate was measured using MTT assay and cell viability; apoptosis, necrosis and late apoptosis and necrosis were evaluated by flow cytometry. The mRNA expression levels of 7 surface marker genes were quantified using RT‐qPCR and CD90 was also analysed by flow cytometry. Differentiation was evaluated using specific staining, measurement of alkaline phosphatase activity and analysis of the mRNA expression. Results: High interindividual differences were observed in proliferation in both cell types, particularly during the final days. Statistically significant differences in viability and early apoptosis of cultured AT‐ and BM‐MSCs were found. The highest values of early apoptosis were observed during the first days of culture, while the highest percentage of necrosis and late apoptosis and lowest viability was observed in the last days. Surface marker expression pattern observed is in accordance to other studies in horse and other species. Osteogenic differentiation was evident after 7 days, with an increasing of ALP activity and mRNA expression of osteogenic markers. Adipogenic differentiation was achieved in BM‐MSCs from 2 donors with one of the 16 media tested. Chondrogenic differentiation was also observed. Conclusions: Proliferation ability is different in AT‐MSCs and BM‐MSCs. Differences in viability and early apoptosis were observed between both sources and CD34 was only found in AT‐MSCs. Differences in their osteogenic and adipogenic potential were detected by staining and quantification of specific tissue markers. Potential relevance: To provide data to better understand AT‐MSCs and BM‐MSCs behaviour in vitro.     

Irrigation water reduction using System of Rice Intensification compared with conventional cultivation methods in Iraq

A field study was conducted at Al-Mishkhab Rice Research Station (MRRS) during the summer season 2009 to evaluate irrigation water use efficiency (IWUE) using Anbar 33 variety with the System of Rice Intensification compared to traditional methods. During the growth phase, the number of leaves, stems, and roots, and the average plant height were measured every 15 days for the two sets of methods. At maturity, the depth and length of plant roots was assessed, along with leaf area index (LAI) of the flag leaf and plant height. The amount of irrigation water applied was measured by water meter for both methods. SRI principles for plant age, spacing, etc., were implemented in the SRI plots. The results indicated more vigorous growth of roots under SRI methods, reaching 13,004 cm plant⁻¹ compared with non-SRI results of 4,722 cm plant⁻¹. There was 42% increase in grain yield when SRI methods were used. These had water use efficiency (WUE) of 0.291 kg m⁻² compared with WUE of 0.108 kg m⁻² for non-SRI cultivation, almost a threefold difference. SRI practices reduced the need for irrigation water by 38.5%.     

Evapotranspiration losses for pepper under plastic mulch and shallow water table conditions

Use of literature crop coefficient (K _c) values for quantifying evapotranspiration (ET_c) under non-standard conditions such as plastic mulch, shallow water table, and sub-tropical conditions can lead to inaccurate ET_c estimates. A 5-year experiment was conducted for fall crop growing seasons in south Florida to quantity bi-weekly ET_c and K _c for bell pepper grown under shallow water table and plastic mulch environments using large drainage lysimeters. The ET_c values varied from 205 to 320 mm with a seasonal average of 267 mm. Average K _c values for bell pepper for development, mid-season, and late stages were 1.05, 1.21, and 1.28, respectively. Higher than literature initial K _c values were due to rainfall and use of sub-irrigation system to maintain artificially high water table which results in high soil moisture in the bare soil area—such high moisture results in high evaporation. The K _c values from this study were statistically higher than literature values. Use of literature K _c values resulted in underestimating ET_c by 27–37%. The K _c values would provide improved estimates of sub-irrigated pepper ET_c in subtropical Florida and elsewhere with similar environment.     

Impact of Organic Amendments on Groundwater Nitrogen Concentrations for Sandy and Calcareous Soils

Experiments were conducted on calcareous and sandy soils to investigate the effects of organic amendments for vegetable production on groundwater nitrogen (N) concentration in south Florida. The treatments consisted of applying yard and food residuals compost, biosolids compost, a cocompost of the municipal solid waste and biosolids, and inorganic fertilizer. Nitrate nitrogen (NO₃-N), ammonium nitrogen (NH₄-N), and total N concentrations were collected for a period of two years for both soils. Statistical analysis results revealed that for the three species tested, there were no significant differences among treatments. NO₃-N concentrations for all treatments remained less than the maximum contamination level (10 mg/L). NO₃-N transport to groundwater was higher in calcareous soil (mean=5.3 mg/L) than in sandy soil (mean=0.6 mg/L). NH₄-N concentrations ranged from 0 to 13.6 mg/L throughout the experiment. Calcareous soil had lower NH₄-N concentrations (mean=0.1 mg/L) than sandy soils (mean=0.7 mg/L). Total N ranged from 0.4 to 21.7 mg/L for all treatments for both soils reflecting high adsorption of dissolved organic N in both soils. Overall, results indicated that all the compost treatments were comparable to inorganic fertilizer with regard to N leaching and N concentrations in the groundwater while producing similar or higher yields.     

Clinicopathological Features of Hemangioma in Two Young Horses

The clinicopathological and immunohistochemical features of two vascular tumors in two young horses are described in the present work. These animals were referred to the Veterinary Teaching Hospital of the University of Córdoba because of the presence of hyperpigmented plaques located in the medial aspect of the left leg and also around the hock (case 1) and in the right front leg (case 2). Some of the lesions showed deep ulceration and severe protrusion with abundant bleeding. The histopathological study revealed that lesions were composed of nonencapsulated, proliferated, closely packed small blood vessels, some of which showed irregular shape, whereas others were similar to capillaries, arterioles, and venules. Neoplastic cells expressed vimentin and factor VIIIar, suggesting their endothelial nature, whereas in the wall of some proliferated vessels, some cells expressed vimentin, α-smooth muscle actin and desmin, an immunophenotype consistent with pericytes, and small muscle cells. These features agree with those reported in human juvenile hemangioma rather than with hemangioma in adult horses.     

Development and evaluation of a loop-mediated isothermal amplification assay for detection of Erwinia amylovora based on chromosomal DNA

A reliable and rapid pathogen detection protocol that utilizes loop-mediated isothermal amplification (LAMP) was developed for detection of Erwinia amylovora, the casual agent of fire blight. The six LAMP primers applied were derived from the highly conserved fragment of the chromosomally amsH gene. Despite the proposed LAMP as well as nested PCR presenting equal values of sensitivity (2?×?10¹?CFU/ml or more) for pure cultures, as compared with conventional PCR (2?×?10³?CFU/ml), both methods were together superior. The specificity assay also showed that the LAMP protocol is species-specific for detection of E. amylovora even in inter-species analysis. Meanwhile, when all 208 naturally infected samples were examined, the specificity value of LAMP was 84%, while conventional and nested PCR could detect only 59% and 73% of the whole collection. Significantly, an independent behaviour versus host plant as well as each strain origin was also observed regarding the current LAMP method as well as other two PCR-based methods. All the results, overall, indicated that the LAMP offers an interesting novel and convenient assay format for the quick and specific chromosomal detection and diagnostic tool of recognition of E. amylovora and therefore presents an alternative to PCR-based assays.     

"Gas and fat embolic syndrome" involving a mass stranding of beaked whales (family Ziphiidae) exposed to anthropogenic sonar signals

A study of the lesions of beaked whales (BWs) in a recent mass stranding in the Canary Islands following naval exercises provides a possible explanation of the relationship between anthropogenic, acoustic (sonar) activities and the stranding and death of marine mammals. Fourteen BWs were stranded in the Canary Islands close to the site of an international naval exercise (Neo-Tapon 2002) held on 24 September 2002. Strandings began about 4 hours after the onset of midfrequency sonar activity. Eight Cuvier's BWs (Ziphius cavirostris), one Blainville's BW (Mesoplodon densirostris), and one Gervais' BW (Mesoplodon europaeus) were examined postmortem and studied histopathologically. No inflammatory or neoplastic processes were noted, and no pathogens were identified. Macroscopically, whales had severe, diffuse congestion and hemorrhage, especially around the acoustic jaw fat, ears, brain, and kidneys. Gas bubble-associated lesions and fat embolism were observed in the vessels and parenchyma of vital organs. In vivo bubble formation associated with sonar exposure that may have been exacerbated by modified diving behavior caused nitrogen supersaturation above a threshold value normally tolerated by the tissues (as occurs in decompression sickness). Alternatively, the effect that sonar has on tissues that have been supersaturated with nitrogen gas could be such that it lowers the threshold for the expansion of in vivo bubble precursors (gas nuclei). Exclusively or in combination, these mechanisms may enhance and maintain bubble growth or initiate embolism. Severely injured whales died or became stranded and died due to cardiovascular collapse during beaching. The present study demonstrates a new pathologic entity in cetaceans. The syndrome is apparently induced by exposure to mid-frequency sonar signals and particularly affects deep, long-duration, repetitive-diving species like BWs.     

An immunohistochemical study of S-100 protein in the intestinal tract of Chinese soft-shelled turtle, Pelodiscus sinensis

The present work describes the distribution of S-100 protein in the intestinal tract of a Chinese soft-shelled turtle specimen (Pelodiscus sinensis). S-100 protein positive cells were located in the intestinal tract, from the proximal small to distal large intestine. S-100 protein positive dendritic cells had irregular shape and were positive in both cytoplasm and nucleus. Most of them were located both lamina propria and submucosa in the small intestine, while few were found in the large intestine. S-100 protein, C-kit positive ICCs and Silver staining glial cells were predominantly observed in three locations: (1) in the interspace between the submucosa and circular muscle layer; (2) in the circular muscle layer; and (3) between the circular and longitudinal muscle layers of the intestine. Fewer were found in the large intestinal lamina propria and submucosa. Three types of positive cells (S-100 protein positive cells, C-kit positive ICCs and Silver staining glial cells) with 1–2 long or 2–3 short processes were distributed as lace-like or surrounding blood vessels in the different locations mentioned above. In the lamina propria, all the positive cells with irregular processes were connected with each other and formed a network. In the submucosa, all the positive cells were found surrounding the blood vessels.     

Physiological responses to water restriction in dry and lactating Awassi ewes

The effect of water restriction in Awassi ewes was assessed under two physiological conditions: lactating and dry, over a 3-week-period (August–September, 2002). Eight dry and eight lactating Awassi ewes (with their lambs) were assigned to one of two watering regimes: watered once every three days and daily watering, respectively. Weather data, body weight and rectal temperature were recorded. Venous blood was sampled on the watering day of the water-restricted group, and subjected to haematological, biochemical and electrolytes analysis. Ewes under water restriction lost more weight and had higher packed cell volume, haemoglobin, serum cholesterol, urea, creatinine, total protein and albumin as compared to daily-watered animals. However, lactation seemed to have no significant effect on these serum components. As for blood pH and electrolytes, Na⁺ and Cl⁻ were significantly higher under water restriction; whereas significantly lower K⁺ and Ca⁺⁺ but higher pH and Cl⁻ were observed in lactating animals. Finally, lower cortisol concentrations were observed in water-restricted animals, warranting further studies on the hormonal adaptation to long term water stress. It was concluded that water restriction induces similar changes in blood physiological indicators in dry and lactating Awassi ewes; while lactation beyond the first month post partum seems to mostly affect blood electrolytes.