Mussel Shells,a Valuable Calcium Resource for the Pharmaceutical Industry

(1) Background: The mussel (Mytilus edulis, Mytilus galloprovincialis) is the most widespread lamellibranch mollusk, being fished on all coasts of the European seas. Mussels are also widely grown in Japan, China, and Spain, especially for food purposes. This paper shows an original technique for mussel shell processing for preparation of calcium salts, such as calcium levulinate. This process involves synthesis of calcium levulinate by treatment of Mytilus galloprovincialis shells with levulinic acid. The advantage of mussel shell utilization results in more straightforward qualitative composition. Thus, the weight of the mineral component lies with calcium carbonate, which can be used for extraction of pharmaceutical preparations. (2) Methods: Shell powder was first deproteinized by calcination, then the mineral part was treated with levulinic acid. The problem of shells generally resulting from the industrialization of marine molluscs creates enough shortcomings, if one only mentions storage and handling. One of the solutions proposed by us is the capitalization of calcium from shells in the pharmaceutical industry. (3) Results: The toxicity of calcium levulinate synthesized from the mussel shells was evaluated by the method known in the scientific literature as the Constantinescu phytobiological method (using wheat kernels, Triticum vulgare Mill). Acute toxicity of calcium levulinate was evaluated; the experiments showed the low toxicity of calcium levulinate. (4) Conclusion: The experimental results highlighted calcium as the predominant element in the composition of mussel shells, which strengthens the argument of capitalizing the shells as an important natural source of calcium.     

Cooperation in private forestry of post-soviet system: Forest owners’ cooperatives in Lithuania

The current state of cooperation in private forestry in Lithuania is examined, with a focus on the analysis of objectives, organisational structure and the ways forest owners’ cooperatives operate. A postal survey has been used as a main research instrument, the questionnaire consisting of a series of multiplechoice close-ended questions. This paper provides insights into the state forest enterprises and other private companies operating in the private forestry sector, and places forest owners’ cooperatives in a broader context of the private forestry sector. A typical forest owner’s cooperative in Lithuania has up to 10 members and about 20 clients to whom services are provided. The leaders of cooperatives indicate that the optimal number of clients using their services should not exceed 40. The main stated objectives of cooperatives are the provision of services to their members under the most attractive conditions, uniting members, and earning a profit for the members. These activities of cooperatives revolve around timber harvesting and trade. It is concluded that the process of cooperation of private forest owners in Lithuania is rather slow, although positive development can be observed.     

First record of autochthonous canine ehrlichiosis caused by Ehrlichia canis in Romania

This case study describes the first genetically confirmed and clinically manifested autochthonous Ehrlichia canis infection in a 9‐year‐old female mixed‐breed dog from Romania. Health screening of the dog included clinical examination, evaluation of stained peripheral blood smear and hematologic variables, as well as serologic testing and molecular analysis. Clinical signs included fever, apathy, dehydration, pale mucous membranes, and weakness. The microscopic examination of the blood smear and immunologic assays for Borrelia burgdorferi, Anaplasma phagocytophilum, and E canis antibodies, and for Dirofilaria immitis antigen yielded negative results. Hematologic abnormalities included moderate nonregenerative anemia, leucopenia with neutropenia, and moderate thrombocytopenia. The biochemical abnormalities identified were hypoalbuminemia, and mildly increased serum enzyme activities of AST and ALT. In addition, increased urea and creatinine levels associated with low urine specific gravity and proteinuria were also present. Nested PCR amplification of the partial E canis 16S rRNA gene demonstrated the presence of this rickettsial pathogen in the dog's blood, which subsequently was confirmed through sequencing based on the 100% homology with GenBank deposited E canis isolates. After specific treatment with doxycycline (10 mg/kg, orally, SID) for one month, the proteinuria, and hematologic and serum biochemical abnormalities with the exception of mild azotemia resolved. This report supports the geographical expansion of canine ehrlichiosis caused by E canis in nonendemic regions of Europe.     

Zero-inflated models for identifying disease risk factors when case detection is imperfect: Application to highly pathogenic avian influenza H5N1 in Thailand

Logistic regression models integrating disease presence/absence data are widely used to identify risk factors for a given disease. However, when data arise from imperfect surveillance systems, the interpretation of results is confusing since explanatory variables can be related either to the occurrence of the disease or to the efficiency of the surveillance system. As an alternative, we present spatial and non-spatial zero-inflated Poisson (ZIP) regressions for modelling the number of highly pathogenic avian influenza (HPAI) H5N1 outbreaks that were reported at subdistrict level in Thailand during the second epidemic wave (July 3rd 2004 to May 5th 2005). The spatial ZIP model fitted the data more effectively than its non-spatial version. This model clarified the role of the different variables: for example, results suggested that human population density was not associated with the disease occurrence but was rather associated with the number of reported outbreaks given disease occurrence. In addition, these models allowed estimating that 902 (95% CI 881–922) subdistricts suffered at least one HPAI H5N1 outbreak in Thailand although only 779 were reported to veterinary authorities, leading to a general surveillance sensitivity of 86.4% (95% CI 84.5–88.4). Finally, the outputs of the spatial ZIP model revealed the spatial distribution of the probability that a subdistrict could have been a false negative. The methodology presented here can easily be adapted to other animal health contexts.     

New histochemical and ultrastructural observations on normal bovine tonsils.

Samples of normal bovine palatine tonsils were examined by light and electron microscopy. Like human tonsils, they were composed of crypts, subepithelial areas, follicles, and T-dependent zones, but their well-developed capsule subdivided the lymphoid tissue by connective septa. B cells formed the major lymphoid component. The follicles and T-dependent zones had morphological and histochemical features typical of peripheral lymph organs. Follicular dendritic cells were isolated and shown to be similar to human follicular dendritic cells.     

Fruiting position during development of ‘Nules Clementine’ mandarin affects the concentration of K,Mg and Ca in the flavedo

Adequate K, Mg and Ca supply is important to develop well-structured and functional cell walls and membranes in fruit, and insufficient levels or imbalances of these minerals are known to be involved in various postharvest disorders. Microclimatic variation exists in the ‘Nules Clementine’ mandarin tree canopy and results in lower photosynthetically active radiation (PAR) levels and temperature as well as a higher humidity inside the tree canopy. The aim of this experiment was to determine the impact of this variation in microclimate on accumulation patterns of K, Ca and Mg in the flavedo of the fruit rind during stages II and III of fruit development. Fruit mass, dimensions, rind colour development and mineral composition of the flavedo were measured to describe the condition of fruit borne on the outside and inside of the tree's canopy. The data revealed that canopy position influenced mineral nutrient accumulation patterns in the flavedo. Outside fruit flavedo accumulated significantly higher concentrations of Ca and Mg in all three seasons (2005–2007). In contrast, inside fruit flavedo (shaded fruit) accumulated significantly higher levels of K compared with outside fruit flavedo. The accumulation of K and Ca differed from that of kiwifruit and apple in that Ca concentration increased and K decreased towards maturity. These results suggest that xylem, as in citrus leaves, is the main vasculature supply conduit to the citrus fruit flavedo for mineral nutrients. The reduction of transpiration potential by lower temperatures and higher humidity inside the canopy could be responsible for the reduced accumulation of Ca and Mg. The high K concentration of inside fruit flavedo is suggested to be a stress response, due to the low light levels, to maintain osmotic potential in the shaded rind tissue, and this imbalance could possibly lead to a reduction in rind condition, which manifests through rind breakdown symptom development.     

Proteomics: A new tool in bovine claw disease research

Claw horn disruption (CHD) is a common underlying cause of lameness in dairy cattle which leads to compromised animal welfare and production losses. Despite an intense research effort over the last two decades, progress in reducing the prevalence of lameness due to CHD has been limited. In addition to current research strategies there is a need to develop novel approaches and methods that expand understanding of the disease mechanisms involved in CHD. The objectives of the present study were to explore the potential of liquid chromatography tandem mass spectrometry (LC-MS/MS) in mapping protein expression in three different bovine claw tissues, and to provide a relevant functional annotation of the proteins characterized in these tissues. LC-MS/MS was used to characterize protein expression in coronary band skin (C), claw dermal (D) and lamellar (L) tissues from two heifers. A total of 388 different proteins were identified, with 146 proteins available for identification in C, 279 proteins in D and 269 proteins in L. A functional annotation of the identified proteins was obtained using the on-line Blast2GO tool. Three hundred and sixteen of the identified proteins could be subsequently grouped manually to one or more of five major functional groups related to metabolism, cell structure, immunity, apoptosis and angiogenesis. These were chosen to represent basic cell functions and biological processes potentially involved in the pathogenesis of CHD. The LC-MS/MS-based proteomic analysis presented here is the largest published survey, so far, of the bovine claw tissue proteome.