Mechanisms for nutrient interactions from organic amendments and mineral fertilizer inputs under cropping systems: a review.

Food security issues continue to be a challenge in most parts of the globe, especially in sub-Saharan Africa (SSA). Several research attempts on addressing this issue have mainly been on nutrient replenishment using combined nutrient application of organic amendments and mineral fertilizer inputs. However, there is limited information available on the potential mechanisms underlying nutrient interactions associated with co-application of organic amendments and mineral fertilizers. Therefore, this review focuses on the mechanisms underlying crop nutrient interactions, with particular emphasis on improved nutrient synchrony, priming effect, general soil fertility improvement and balanced proportion of nutrients required by crops. Following a brief overview of the mechanisms, the review describes four common pre-determined nutrient ratios required by plants depending on its life cycle, environment and genotypic characteristics in order to attain the crop's maximum genetic potential. The review concludes with the need for future research to understudy mechanisms causing nutrient interaction under cropping systems, so as to apply nutrients at the most appropriate time to synchronize nutrient release with crop uptake, with the utmost goal of promoting sustainable crop production and enhancing food security.

Economically Optimal Rate for Nutrient Application to Maize in the Semi-deciduous Forest Zone of Ghana.

Low inherent nitrogen (N), phosphorus (P), and potassium (K) contents of smallholder farms limit maize grain yield. Maize grain yield response to N, P, and K mineral fertilizer application and economically optimal rates for nitrogen (EORN), phosphorus (EORP), and potassium (EORK) were evaluated on a Ferric Acrisol within the semi-deciduous forest zone of Ghana. The nutrient rates evaluated were N (0, 30, 60, 90, and 120 kg N ha-1), P (0, 30, 60, and 90 kg ha-1 P2O5), and K (0, 30, 60 and 90 kg ha-1 K2O). The treatments were arranged in a randomized complete block with three replications using an incomplete factorial design. Nutrient responses were determined using asymptotic quadratic-plus plateau functions. The best nitrogen rate for all P and K levels was 60 kg ha-1, which gave grain yield of 5 t ha-1 . Nitrogen uptake, N agronomic and N recovery efficiencies peaked at 60 kg N ha-1 while N partial factor productivity declined with increasing N application rate. Cost to grain price ratios (CP) were 1.29, 1.65, and 1.65 for N, P, and K, respectively. The EORN was 61 kg ha-1, 32% less than the recommended 90 kg N ha-1 for maize production in the semi-deciduous forest zone of Ghana. Nitrogen application had the lowest CP ratio, making its application economically profitable than P and K. The findings suggest that the application of N at 61 kg N ha-1 to maize is economically profitable than at higher application rates. However, further studies should be conducted on farmers' fields to validate the results obtained.

Biosafety Evaluation of Equine Umbilical Cord-Derived Mesenchymal Stromal Cells by Systematic Pathogen Screening in Peripheral Maternal Blood and Paired UC-MSCs.

Background: The growing interest in mesenchymal stromal cells (MSCs) in equine medicine, together with the development of MSC biobanking for allogeneic use, raises concerns about biosafety of such products. MSCs derived from umbilical cord (UC) carry an inherent risk of contamination by environmental conditions and vertical transmission of pathogens from broodmares. There is yet no report in the scientific literature about horses being contaminated by infected MSC products, and no consensus about systematic infectious screening of umbilical cord-derived mesenchymal stromal cells (UC-MSCs) to ensure microbiological safety of therapeutic products. Objectives: To develop a standard protocol to ensure UC-MSC microbiological safety and to assess the risk of vertical transmission of common intracellular pathogens from broodmares to paired UC-MSCs. Study Design and Methods: Eighty-four UC and paired peripheral maternal blood (PMB) samples were collected between 2014 and 2016. Sterility was monitored by microbiological control tests. Maternal contamination was tested by systematical PMB PCR screening for 14 pathogens and a Coggins test. In case of a PCR-positive result regarding one or several pathogen(s) in PMB, a PCR analysis for the detected pathogen(s) was then conducted on the associated UC-MSCs. Results: Ten out of 84 UC samples were contaminated upon extraction and 6/84 remained positive in primo culture. The remaining 78/84 paired PMB & UC-MSC samples were evaluated for vertical transmission; 37/78 PMB samples were PCR positive for Equid herpesvirus (EHV)-1, EHV-2, EHV-5, Theileria equi, Babesia caballi, and/or Mycoplasma spp. Hepacivirus was detected in 2/27 cases and Theiler Diseases Associated Virus in 0/27 cases (not performed on all samples due to late addition). All paired UC-MSC samples tested for the specific pathogen(s) detected in PMB were negative (37/37). Main Limitations: More data are needed regarding MSC susceptibility to most pathogens detected in PMB. Conclusions: In-process microbiological controls combined with PMB PCR screening provide a comprehensive assessment of UC-MSC exposure to infectious risk, vertical transmission risk appearing inherently low.

Long-Term Safety and Efficacy of Single or Repeated Intra-Articular Injection of Allogeneic Neonatal Mesenchymal Stromal Cells for Managing Pain and Lameness in Moderate to Severe Canine Osteoarthritis Without Anti-inflammatory Pharmacological Support: Pilot Clinical Study.

Objective: To explore the long-term safety and efficacy of canine allogeneic mesenchymal stromal cells (MSC) administered intra-articularly as single or repeated injections in appendicular joints of dogs affected by moderate to severe refractory osteoarthritis. Study Design: 22 pet dogs were recruited into a non-randomized, open and monocentric study initially administering one cellular injection. A second injection was offered after 6 months to owners if the first injection did not produce expected results. Materials and Methods: Anti-inflammatory treatment (if prescribed) was discontinued at last one week before the onset of treatment. Each injection consisted of at least 10 million viable neonatal allogeneic mesenchymal stromal cells obtained from fetal adnexa. Medical data was collected from veterinary clinical evaluations of joints up to 6 months post-injection and owner's assessment of their dog's mobility and well-being followed for a further 2 years when possible. Results: Mild, immediate self-limiting inflammatory joint reactions were observed in 5/22 joints after the first injection, and in almost all dogs having a subsequent injection. No other MSC-related adverse medical events were reported, neither during the 6 months follow up visits, nor during the long-term (2-years) safety follow up. Veterinary clinical evaluation showed a significant and durable clinical improvement (up to 6 months) following MSC administration. Eight dogs (11 joints) were re-injected 6 months apart, sustaining clinical benefits up to 1 year. Owner's global satisfaction reached 75% at 2 years post-treatment Conclusion: Our data suggest that a single or repeated intra-articular administration of neonatal MSC in dogs with moderate to severe OA is a safe procedure and confer clinical benefits over a 24-month period. When humoral response against MSC is investigated by flow cytometry, a positive mild and transient signal was detected in only one dog from the studied cohort, this dog having had a positive clinical outcome.

On-farm evaluation and determination of sources of variability of soybean response to Bradyrhizobium inoculation and phosphorus fertilizer in northern Ghana.

Soybean yields on smallholder farms in sub-Sahara Africa (SSA) are far below the potential yield thus creating a huge yield gap. Interventions are thus needed to bridge this yield gap and ascertain the factors influencing the yield variation. This study evaluated the on farm response of soybean to rhizobia inoculation and or mineral P fertilizer in Northern and Upper West regions of Ghana in a single non-replicate trial using four treatments: no input (control), TSP fertilizer (P), rhizobia inoculant (I) and TSP plus inoculant (P + I). In addition, the study sought to develop a robust approach for determining responsiveness and non-responsiveness using agronomic and economic indices. The results showed that the average grain yield of plots that received P or I were higher than control plots. Higher grain yield responses were however, obtained by the plots that received combined application of P and Bradyrhizobium inoculant. Grain yield response in the Northern region was higher than in the Upper West region. Response to P and or I were highly variable within and between locations. The cumulative rainfall and some soil factors including soil nitrogen, phosphorus, soil type, organic carbon, pH and texture explained about 42-79% of these variations in soybean grain yield. The agronomic approach for determining responsive and non-responsiveness revealed that 17-40 % and 6-17% of the locations within the Northern and Upper West regions, respectively were responsive to P fertilization and/ or Bradyrhizobium inoculation. However, the economic approach indicated that 64-75% and 14-24% of the locations within the Northern and Upper West regions, respectively were responsive to P fertilization and Bradyrhizobium inoculation. The results imply that rhizobia inoculation is an effective strategy for increasing soybean yield and improving livelihood of smallholder farmers.

Occurrence and risk assessment of antibiotics in water and lettuce in Ghana.

Hospital wastewater and effluents from waste stabilization ponds in Kumasi, Ghana, are directly discharged as low quality water into nearby streams which are eventually used to irrigate vegetables. The presence of 12 commonly used antibiotics in Ghana (metronidazole, ciprofloxacin, erythromycin, trimethoprim, ampicillin, cefuroxime, sulfamethoxazole, amoxicillin, tetracycline, oxytetracycline, chlortetracycline and doxycycline) were investigated in water and lettuce samples collected in three different areas in Kumasi, Ghana. The water samples were from hospital wastewater, wastewater stabilization ponds, rivers and irrigation water, while the lettuce samples were from vegetable farms and market vendors. Antibiotics in water samples were extracted using SPE while antibiotics in lettuce samples were extracted using accelerated solvent extraction followed by SPE. All extracted antibiotics samples were analyzed by HPLC-MS/MS. All studied compounds were detected in concentrations significantly higher (p=0.01) in hospital wastewater than in the other water sources. The highest concentration found in the present study was 15µg/L for ciprofloxacin in hospital wastewater. Irrigation water samples analyzed had concentrations of antibiotics up to 0.2µg/L. Wastewater stabilization ponds are low technology but effective means of removing antibiotics with removal efficiency up to 95% recorded in this study. However, some chemicals are still found in levels indicating medium to high risk of antibiotics resistance development in the environment. The total concentrations of antibiotics detected in edible lettuce tissues from vegetable farms and vegetable sellers at the markets were in the range of 12.0-104 and 11.0-41.4ng/kg (fresh weight) respectively. The antibiotics found with high concentrations in all the samples were sulfamethoxazole, erythromycin, ciprofloxacin, cefuroxime and trimethoprim. Furthermore, our study confirms the presence of seven antibiotics in lettuce from irrigation farms and markets, suggesting an indirect exposure of humans to antibiotics through vegetable consumption and drinking water in Ghana. However, estimated daily intake for a standard 60kg woman was 0.3ng/day, indicating low risk for human health.

Evaluation of the Effect of a Single Intra-articular Injection of Allogeneic Neonatal Mesenchymal Stromal Cells Compared to Oral Non-Steroidal Anti-inflammatory Treatment on the Postoperative Musculoskeletal Status and Gait of Dogs over a 6-Month Period after Tibial Plateau Leveling Osteotomy: A Pilot Study.

OBJECTIVE: Compare the clinical and pressure walkway gait evolution of dogs after a tibial plateau leveling osteotomy (TPLO) for a cranial cruciate ligament rupture (CrCLR) and treatment with either a 1-month course of non-steroidal anti-inflammatory drugs (NSAIDs) or a single postoperative intra-articular (IA) injection of allogeneic neonatal mesenchymal stromal cells (MSCs). STUDY DESIGN: Prospective, double-blinded, randomized, controlled, monocentric clinical study. ANIMALS: Sixteen client-owned dogs. MATERIALS AND METHODS: Dogs with unilateral CrCLR confirmed by arthroscopy were included. Allogeneic neonatal canine MSCs were obtained from fetal adnexa retrieved after C-section performed on healthy pregnant bitches. The dogs were randomly allocated to either the "MSCs group," receiving an IA injection of MSCs after TPLO, followed by placebo for 1 month, or the "NSAIDs group," receiving IA equivalent volume of MSCs vehicle after TPLO, followed by oral NSAID for 1 month. One of the three blinded evaluators assessed the dogs in each group before and after surgery (1, 3, and 6 months). Clinical score and gait and bone healing process were assessed. The data were statistically compared between the two groups for pre- and postoperative evaluations. RESULTS: Fourteen dogs (nine in the MSCs group, five in the NSAIDs group) completed the present study. No significant difference was observed between the groups preoperatively. No local or systemic adverse effect was observed after MSCs injection at any time point considered. At 1 month after surgery, bone healing scores were significantly higher in the MSCs group. At 1, 3, and 6 months after surgery, no significant difference was observed between the two groups for clinical scores and gait evaluation. CONCLUSION: A single IA injection of allogeneic neonatal MSCs could be a safe and valuable postoperative alternative to NSAIDs for dogs requiring TPLO surgery, particularly for dogs intolerant to this class of drugs.

Tetraspanin 8 is a novel regulator of ILK-driven ß1 integrin adhesion and signaling in invasive melanoma cells.

Melanoma is well known for its propensity for lethal metastasis and resistance to most current therapies. Tumor progression and drug resistance depend to a large extent on the interplay between tumor cells and the surrounding matrix. We previously identified Tetraspanin 8 (Tspan8) as a critical mediator of melanoma invasion, whose expression is absent in healthy skin. The present study investigated whether Tspan8 may influence cell-matrix anchorage and regulate downstream molecular pathways leading to an aggressive behavior. Using silencing and ectopic expression strategies, we showed that Tspan8-mediated invasion of melanoma cells resulted from defects in cell-matrix anchorage by interacting with ß1 integrins and by interfering with their clustering, without affecting their surface or global expression levels. These effects were associated with impaired phosphorylation of integrin-linked kinase (ILK) and its downstream target Akt-S473, but not FAK. Specific blockade of Akt or ILK activity strongly affected cell-matrix adhesion. Moreover, expression of a dominant-negative form of ILK reduced ß1 integrin clustering and cell-matrix adhesion. Finally, we observed a tumor-promoting effect of Tspan8 in vivo and a mutually exclusive expression pattern between Tspan8 and phosphorylated ILK in melanoma xenografts and human melanocytic lesions. Altogether, the in vitro, in vivo and in situ data highlight a novel regulatory role for Tspan8 in melanoma progression by modulating cell-matrix interactions through ß1 integrin-ILK axis and establish Tspan8 as a negative regulator of ILK activity. These findings emphasize the importance of targeting Tspan8 as a means of switching from low- to firm-adhesive states, mandatory to prevent tumor dissemination.

Contribution of Wastewater Irrigation to Soil Transmitted Helminths Infection among Vegetable Farmers in Kumasi, Ghana.

Wastewater irrigation is associated with several benefits but can also lead to significant health risks. The health risk for contracting infections from Soil Transmitted Helminths (STHs) among farmers has mainly been assessed indirectly through measured quantities in the wastewater or on the crops alone and only on a limited scale through epidemiological assessments. In this study we broadened the concept of infection risks in the exposure assessments by measurements of the concentration of STHs both in wastewater used for irrigation and the soil, as well as the actual load of STHs ova in the stool of farmers and their family members (165 and 127 in the wet and dry seasons respectively) and a control group of non-farmers (100 and 52 in the wet and dry seasons, respectively). Odds ratios were calculated for exposure and non-exposure to wastewater irrigation. The results obtained indicate positive correlation between STH concentrations in irrigation water/soil and STHs ova as measured in the stool of the exposed farmer population. The correlations are based on reinfection during a 3 months period after prior confirmed deworming. Farmers and family members exposed to irrigation water were three times more likely as compared to the control group of non-farmers to be infected with Ascaris (OR = 3.9, 95% CI, 1.15-13.86) and hookworm (OR = 3.07, 95% CI, 0.87-10.82). This study therefore contributes to the evidence-based conclusion that wastewater irrigation contributes to a higher incidence of STHs infection for farmers exposed annually, with higher odds of infection in the wet season.

The policy implications of urban open space commercial vegetable farmers' willingness and ability to pay for reclaimed water for irrigation in Kumasi, Ghana.

The acute waste management problems, coupled with the proliferation of small scale industries in many developing countries, make low quality water treatment before use inevitable in the long run. These industries have the potential to discharge effluent containing chemicals and heavy metals into the environment. The indiscriminative use of pharmaceutical products by households in many of these countries is another source of health concern. Low quality water treatment in these countries has however been hampered by the high cost of infrastructure provision and maintenance. Cost-sharing among stakeholders appears to be a promising strategy to finance and maintain the wastewater treatment infrastructure. In this study therefore, the willingness and ability of urban open space commercial vegetable farmers to pay for reclaimed water for irrigation purposes has been assessed. One hundred open space commercial vegetable farmers and four vegetable farmers' associations were selected and interviewed in Kumasi in Ghana using semi-structured interview schedules and interview guides respectively. The results of the study show that approximately three out of every five vegetable farmers were willing to pay for reclaimed water for irrigation. The results further show that the probability of being willing to pay by farmers who agreed that the current water they used for irrigation was harmful is approximately 5.3 times greater than that of those who did not. The analysis of the farmers' ability to pay revealed that all the farmers would be capable of paying for reclaimed water at a price of US$0.11/m(3). This has implications for land tenure security and vegetable consumers' willingness to pay higher prices for the produce.

Uptake of antibiotics from irrigation water by plants.

The capacity of carrot (Daucus corota L.) and lettuce (Lactuca sativa L.), two plants that are usually eaten raw, to uptake tetracycline and amoxicillin (two commonly used antibiotics) from irrigated water was investigated in order to assess the indirect human exposure to antibiotics through consumption of uncooked vegetables. Antibiotics in potted plants that had been irrigated with known concentrations of the antibiotics were extracted using accelerated solvent extraction and analyzed on a liquid chromatograph-tandem mass spectrometer. The plants absorbed the antibiotics from water in all tested concentrations of 0.1-15 mg L(-1). Tetracycline was detected in all plant samples, at concentrations ranging from 4.4 to 28.3 ng/g in lettuce and 12.0-36.8 ng g(-1) fresh weight in carrots. Amoxicillin showed absorption with concentrations ranging from 13.7 ng g(-1) to 45.2 ng g(-1) for the plant samples. The mean concentration of amoxicillin (27.1 ng g(-1)) in all the samples was significantly higher (p = 0.04) than that of tetracycline (20.2 ng g(-1)) indicating higher uptake of amoxicillin than tetracycline. This suggests that the low antibiotic concentrations found in plants could be important for causing antibiotics resistance when these levels are consumed.

Canine placenta: A promising potential source of highly proliferative and immunomodulatory mesenchymal stromal cells?

In veterinary medicine, therapeutic mesenchymal stromal cells (MSC) have been traditionally isolated from adult bone marrow or adipose tissue. Neonatal tissues, normally discarded at birth from all species have become an alternative source of cells for regenerative medicine in the human clinic. These cells have been described as being more primitive, proliferative and immunosuppressive than their adult counterparts. Our objective was to examine if this phenomena holds true in dogs. Little information exists regarding canine neonatal MSC characterisation. In this study, we were able to both isolate, phenotype and assess the differentiation and immunomodulatory properties of MSC from canine foetal adnexa allowing us to compare their characteristics to their more well-known bone marrow (BM) cousins. Neonatal tissues, including amnion (AM), placenta (PL), and umbilical cord matrix (UCM) were collected from 6 canine caesarean sections. Primary cells were expanded in vitro for 5 consecutive passages and their proliferation measured. BM-MSC were isolated from 5 control dogs euthanised from other studies and grown in vitro using an identical protocol. All MSC lines were systematically evaluated for their ability to differentiate into 3 mesodermal lineages (adipocyte, osteocyte and chondrocyte) and phenotyped by cytometry and qPCR. In addition, the enzymatic activity of the key immunomodulatory marker indoleamine 2,3-dioxygenase (IDO) was evaluated for each MSC line. MSC displaying a fibroblastic appearance were successfully grown from all neonatal tissues. PL-MSC exhibited significantly higher proliferation rates than AM- and UCM-MSC (p=0.05). Cytometric analysis showed that all MSC express CD90, CD29, and CD44, while no expression of CD45, CD34 and MHC2 was detected. Molecular profiling showed expression of CD105 and CD73 in all MSC. Low levels of SOX2 mRNA was observed in all MSC, while neither NANOG, nor OCT4 were detected. All MSC differentiate into 3 mesodermal lineages. Following inflammatory stimulation, the activity of the immunomodulatory enzyme IDO was significantly higher in neonatal MSC compared to BM-MSC (p=0.009). Our results show that canine foetal adnexa cells share very similar properties to their adult equivalents but upon stimulation show significantly higher IDO immunomodulatory activity. Further studies will be needed to confirm the potential therapeutic benefits of these cells.

The usefulness of the validated SkinEthic™ RHE test method to identify skin corrosive UN GHS subcategories.

The SkinEthic™ Reconstructed Human Epidermis (RHE) test method has been adopted within the context of OECD TG 431 for distinguishing corrosive and non-corrosive chemicals. The EU CLP classification system requires subcategorising of corrosive chemicals into the three UN GHS subcategories 1A, 1B and 1C. Since the SkinEthic™ RHE method was originally validated to discriminate corrosives from non-corrosives, the present study was undertaken to investigate its usefulness to discriminate skin corrosive UN GHS subcategories. In total 84 substances were tested in three independent runs and two prediction models (PM) were assessed, representing a pre-defined validated prediction model (PM-A) and an alternative one defined post-hoc (PM-B). The results obtained with both PM were reproducible, as shown by the ⩾92.9% concordance of classification between runs for discriminating corrosives versus non-corrosives, and the ⩾85% concordance for discriminating the GHS subcategories versus non-corrosives. Moreover results confirmed a high sensitivity of the SkinEthic™ RHE method to predict corrosives (94.9%) and good specificity (⩾73.7%) independent of the PM applied. Regarding the identification of UN GHS corrosive subcategories, PM-A resulted in 86.1% correct classifications of the GHS subcategory 1A. When using the PM-B, the identification of GHS subcategory 1B-and-1C substances improved, with 63.4% correct sub-categorisation. If considering the 30 reference chemicals as recommended in the recently revised OECD TG 431 (2013), PM-A and PM-B achieved 78.9% and 83.3% accuracy respectively for the identification of GHS subcategories and non-corrosives. They correctly predicted 90% of GHS subcategory 1A and 80% of GHS non-corrosive substances independent of the PM used. In conclusion, the SkinEthic™ RHE test method is highly reproducible and sensitive for discriminating corrosive from non-corrosive substances. Furthermore it allows reliable identification of skin corrosive GHS subcategory 1B-and-1C substances using the PM-A and PM-B, and of GHS subcategories 1A using the PM-B. Due to its high sensitivity, the test method provides high safety standards for skin corrosion testing.

Aluminum resistance of cowpea as affected by phosphorus-deficiency stress.

Plants growing in acid soils suffer both phosphorus (P) deficiency and aluminum (Al) toxicity stresses. Selection of genotypes for adaptation to either P deficiency or Al toxicity has sometimes been unsuccessful because these two soil factors often interact. Two experiments were conducted to evaluate eight cowpea genotypes for Al resistance and to study the combined effect of P deficiency and Al toxicity stress on growth, P uptake, and organic acid anion exudation of two genotypes of contrasting Al resistance selected from the first experiment. Relative root inhibition by 30 microM Al ranged from 14% to 60% and differed significantly among the genotypes. Al significantly induced callose formation, particularly in Al-sensitive genotypes. P accumulation was significantly reduced (28% and 95%) by Al application for both the Al-resistant and the Al-sensitive genotypes. Al supply significantly enhanced malate release of root apices of both genotypes. However, the exudation rate was significantly higher in the Al-resistant genotype. P deprivation induced an enhanced malate exudation in the presence of Al only in the Al-resistant genotype IT89KD-391. Citrate exudation rate of the root apices was lower than malate exudation by a factor of about 10, and was primarily enhanced by P deficiency in both genotypes. Al treatment further enhanced citrate exudation in P-sufficient, but not in P-deficient plants. The level of citrate exudation was consistently higher in the Al-resistant genotype IT89KD-391 particularly in presence of Al. It is concluded that the Al-resistant genotype is better adapted to acid Al-toxic and P-deficient soils than the Al-sensitive genotype since both malate and citrate exudation were more enhanced by combined Al and P-deficiency stresses.