Changes of Cytochemical Markers in the Conjunctival and Corneal Epithelium After Corneal Debridement

1. The aim of this study was to determine the epithelial changes of the conjunctiva and cornea up to 7 days after corneal debridement and the changes highlighted included (1) proliferation, (2) production of growth factor, (3) changes in calcium binding protein marker, (4) production of cytokine, and (5) maturity of the regeneration corneal epithelium.2. The cytochemical changes of the corneal and conjunctival epithelia of rabbit were analyzed up to 7 days after debridement.3. An increase in proliferating cell nuclear antigen (PCNA) was observed in the limbal epithelia 12 hr after lesion and reached a peak by 48 hr.4. Some proliferating limbal cells also contained epidermal growth factor (EGF) beginning 24 hr after injury. The early limbal cell proliferation and the EGF production and their persistence until 7 days after lesion were likely involved with the process of regeneration.5. Other positive markers appeared after lesion included tumor necrosis factor (TNF) and calcium binding proteins S100A and S100B, which appeared mainly within the first 48 hr after lesion and then started to decline. The short appearance and the relatively small quantity of TNF indicated that this cytokine was probably not very important in the repair process and its appearance might be related to the injury induced. The presence of S100A and S100B could be associated with both cell death after injury and the proliferation of new epithelium.6. The cornea epithelium was still immature 7 days after lesion in that it still contained cytokeratin.7. In conclusion, the critical hours of peak conjunctival and corneal changes after corneal debridement were in the first 2 days.     

Soil macrofauna diversity as a key element for building sustainable agriculture in Argentine Pampas

The agricultural activity in the Argentine Pampas, characterized by an important trend towards no-till soybean monocropping, has completely transformed the original Pampas landscape into a monotonous scenario with a continuous succession of farms of very low crop diversity. This process has led to soil physical, chemical and biological degradation in those systems. The increase of crop rotation rates in no-till and reduced tillage systems has been proposed as an alternative with reduced negative impact on soils in the context of conventional agriculture. On the other hand, extensive organic farming is also suggested as an alternative to high-input agriculture systems. In this article, we aim to explore how different variations of farming practices and systems impact soil macrofauna, along an edaphoclimatic gradient in the Pampas region. We studied the following systems: natural grassland (Gr) as indicator of the original community, extensive organic farming (Org), conventional agriculture with no-tillage and three crop rotation levels (Nt-R1, Nt-R2 and Nt-R3), and reduced tillage with two levels of crop rotation (Til and Til-R). We assessed soil macrofauna, with emphasis on earthworm, beetle and ant communities; and soil physical and chemical properties. Macrofaunal taxa composition was significantly affected by both management systems and edaphoclimatic conditions. The Gr community had pronounced differences from all the agricultural systems. The earthworm community from Gr had distinctive features from those of most agricultural systems, with Org and Nt-R3 being the most similar to Gr in native and exotic earthworm species, respectively. The beetle community in Org was the most different one, and the communities from the other systems did not show a pattern related to management. Ant community composition was not determined by management systems, but it was affected by edaphoclimatic conditions. All the studied macrofauna groups had a significant co-variation with soil physical and chemical properties, showing that both the characteristics of each soil relative to the geographic location and the effect of management on abiotic soil attributes have an important effect on soil macrofauna. This study confirms that biodiversity is being lost in Pampas soils, which implies a possible threat to the soil capacity to perform the processes that sustain soil functioning and hence plant productivity. Further considerations about the sustainability of the current agricultural model applied in the Argentine Pampas are needed.     

Quality control of a medicinal larval (Lucilia sericata) debridement device based on released gelatinase activity

Lucilia sericata Meigen (Diptera: Calliphoridae) larvae are manufactured worldwide for the treatment of chronic wounds. Published research has confirmed that the primary clinical effect of the product, debridement (the degradation of non‐viable wound tissue), is accomplished by a range of enzymes released by larvae during feeding. The quality assessment of larval activity is currently achieved during production using meat‐based assays, which monitor insect growth and/or the reduction in substrate mass. To support this, the present authors developed a complementary radial diffusion enzymatic assay to produce a visual and measureable indication of the activity of larval alimentary products (LAP) collected under standardized conditions, against a gelatin substrate. Using basic laboratory equipment and reagents, the assay is rapid and suited to high throughput. Assay reproducibility is high (standard deviation: 0.06–0.27; coefficient of variation: 0.75–4.31%) and the LAP collection procedure does not adversely affect larval survival (mortality: < 2%). Because it is both cost‐ and time‐effective, this method is suited to both academic and industrial use and supports good manufacturing and laboratory practice as a quality control assay.     

A new method for the production of sterile colonies of Lucilia sericata

Maggot debridement therapy (MDT) refers to the use of blowfly larvae to clean or debride an infected wound. Most commonly, larvae of Lucilia sericata (Meigen) (Diptera: Calliphoridae) are used, and are sterilized prior to use to ensure no further bacterial infections are introduced during treatment. Current methods sterilize eggs from laboratory‐reared blowfly colonies, after which sterile early second instar maggots can be provided to hospitals for use in treatment. Maggots not required for treatment are used for colony regeneration, in which sterility is not maintained. The ability to maintain sterility beyond this would allow further research into fly–bacteria interactions and the effects of different bacteria on the blowfly lifecycle. This study aimed to produce a colony of sterile adults, using current egg sterilization practice, but maintaining sterility through to pupation and emergence. The production of a sterile colony allows further research into the impact of bacteria on fly development and survival. Eggs were placed on a sterile food source within autoclaved plant tissue culture containers to allow growth under sterile conditions. Nutrient agar plating of sterilized and non‐sterilized eggs, larvae and adults (post‐emergence), as well as the pupation medium and feed source in nutrient broth confirmed the aerobic sterility of all samples involved. The lifecycle of L. sericata was successfully completed through pupation to emergence with no effects on lifespan or oviposition by the newly emerged, sterile adult colony.     

The maggot therapy supply chain: a review of the literature and practice

Maggot therapy (MT) is the clinical application of living fly larvae for the treatment of non-healing wounds and wounds that require debridement. This systematized and expanded literature review is the first study to investigate MT through the conceptual and disciplinary framework of supply chain management. The review of 491 selected academic papers was expanded to include the grey literature and online information resources to construct a first-pass theory of the medicinal maggot supply chain. It shows that the literature to date has focused on isolated discussions of echelon-specific issues such as diet improvement and sterilization protocols in the production echelon, and the relative effectiveness of medicinal maggot application methods in the treatment echelon. There is little knowledge in the public domain regarding the transport and distribution of medicinal maggots, but existing supply chains for vaccines, blood and pathology specimens may provide learning and supply chain integration opportunities. Maggot therapy knowledge across the treatment echelon is generally substantive but there is still insufficient knowledge regarding patients' and health care providers' attitudes toward the therapy, and their experiences of receiving and administering MT. Moreover, there is no research concerned with the humane disposal of medicinal flies during production and after treatment.     

The influence of environmental factors and management methods on the vegetation of mesic grasslands in a central European mountain range

We examined how environmental factors combined with the diverse ways of the extensive management of mesic mountain grasslands affect species composition and diversity in the Polish part of Central Sudetes Mts. Based on the data from 100 research plots altitude, organic matter content and exchangeable magnesium in the soil, as well as the maximum soil water capacity and amount of sand fraction significantly affected species composition. Among the management methods, a significant effect on the differentiation of species composition was observed following the cessation of usage and mowing. Mown meadows had the highest share of forbs in the biomass, whereas those abandoned ones were dominated by tall and expansive grasses, mainly Calamagrostis epigejos. The species richest grasslands occurred in areas with soils of high water capacity, containing high concentrations of calcium and low amounts of total nitrogen. No significant effect of management methods on the total number of species as well as on the number of forb species was observed. Mown pastures had the highest mean value of the Shannon–Wiener diversity index.     

Green tissue within the haustorium of the dwarf mistletoe Korthalsella (Viscaceae). An ultrastructural comparison between chloroplasts of sucker and aerial stem tissues

Summary Korthalsella (Viscaceae) is a dwarf mistletoe attached to its host branch by a single haustorium. Plants are leafless with flattened or cylindrical stems that function in photosynthesis. When a fresh haustorium is cut the sucker within the host appears bright green. Transmission electron microscopy reveals that this greening is due to chloroplasts, but that their organization differs from those of the aerial stem. The three representatives of Korthalsella endemic to New Zealand were the main species investigated. In the stem, chloroplasts have short stacks of cylindrical grana interconnected by stroma thylakoids typical of normal chloroplasts. Sucker chloroplasts have a more variable organization, with most containing extensive granal stacks and poorly differentiated stroma thylakoids. These granal thylakoids exhibit extensive partitions formed by appression of adjacent membranes. Some sucker plastids also approach etioplasts in having a prominent prolamellar body from which radiate thylakoids with short partitions. Sucker chloroplasts usually contain a few large starch grains, plastoglobuli, and sometimes also a stroma centre. The extensive granal thylakoids in sucker chloroplasts of Korthalsella resemble that found in certain shade plants and tissue grown under low light conditions. Sucker chloroplasts probably have a low level of photosynthesis. This activity might provide a local source of osmotically active material used to assist transport between host and parasite.     

A simple,heat‐sterilizable artificial diet excluding animal‐derived ingredients for adult blowfly,Lucilia sericata

Artificial diets have been developed for Lucilia sericata (Meigen) blowfly larvae; however, diets for adults have not yet been developed. An adult diet that excludes animal tissues and animal‐derived ingredients and promotes not only ovarian development, but also oviposition, would aid in basic research and maggot debridement therapy. We have successfully developed artificial diets that exclude animal tissues and animal‐derived ingredients for L. sericata adults. The outcomes of the diets were comparable with those of a beef liver diet in terms of oviposition, adult survival and number of offspring.