Evaluation of the phytoestrogenic activity of Cyclopia genistoides (honeybush) methanol extracts and relevant polyphenols

Unfermented C. genistoides methanol extracts of different harvestings and selected polyphenols were evaluated for phytoestrogenic activity by comparing binding to both ER subtypes, transactivation of an ERE-containing promoter reporter, proliferation of MCF-7-BUS and MDA-MB-231 breast cancer cells, and binding to SHBG. The extracts from one harvesting of C. genistoides (P104) bound to both ER subtypes. All extracts transactivated ERE-containing promoter reporters via ERbeta but not via ERalpha. All extracts, except P122, caused proliferation of the estrogen-sensitive MCF-7-BUS cells. Proliferation of MCF-7-BUS cells was ER-dependent as ICI 182,780 reversed proliferation. Physiologically more relevant, extracts antagonized E2-induced MCF-7-BUS cell proliferation. Furthermore, all extracts, except P122, induced proliferation of the estrogen-insensitive MDA-MB-231 cells, suggesting that the extracts are able to induce ER-dependent and ER-independent cell proliferation. Binding to SHBG by extracts was also demonstrated. These results clearly show that C. genistoides methanol extracts display phytoestrogenic activity and act predominantly via ERbeta. HPLC and LC-MS analysis, however, suggests that the observed phytoestrogenic activity cannot be ascribed to polyphenols known to be present in other Cyclopia species.     

Surveys on Coxiella burnetii infections in Swedish cattle,sheep, goats and moose

Background

Q fever is a zoonotic disease caused by the bacterium Coxiella burnetii. Prevalence data in ruminant species are important to support risk assessments regarding public and animal health. The aim was to investigate the presence of or exposure to C. burnetii in cattle, sheep, goats and moose, and to compare two enzyme-linked immunosorbent assays (ELISAs). National surveys of antibodies against C. burnetii were performed for dairy cattle (n=1537), dairy goats (n=58) and sheep (n=518). Bovine samples consisted of bulk milk, caprine of pooled milk, and ovine of pooled serum. Antibodies were investigated in moose samples (n=99) from three regions. A one-year regional cattle bulk milk survey was performed on the Isle of Gotland (n=119, four occasions). Cattle, sheep and goat samples were analysed with indirect ELISA and moose samples with complement fixation test. For the sheep, goat, and parts of the cattle survey, samples were run in parallel by ELISAs based on antigens from infected ruminants and ticks. Bulk milk samples from the regional cattle survey and vaginal swabs from a subset of the sheep herds (n=80) were analysed for the agent by polymerase chain reaction. Spatial clustering was investigated in the national cattle survey.

Results

The prevalence of antibodies in dairy herds was 8.2% with large regional differences. High risk clusters were identified in the southern regions. The prevalence among dairy herds on the Isle of Gotland varied from 55.9% to 64.6% and 46.4% to 58.9.0% for antibodies and agent, respectively, overall agreement between agent and antibodies was 85.2%. The prevalence of antibodies in sheep was 0.6%, the agent was not detected the vaginal swabs. Antibodies were not detected in goats or moose, although parts of the moose samples were collected in an area with high prevalence in cattle. The overall agreement between the two ELISAs was 90.4%.

Conclusions

The prevalence of antibodies against C. burnetii in dairy cattle in Sweden shows large regional differences. The results suggest that C. burnetii is a rare pathogen among Swedish moose, dairy goat and sheep. ELISAs based on ruminant and tick antigen performed in a similar manner under Swedish conditions.     

Associations between bacterial genotype and outcome of bovine clinical Staphylococcus aureus mastitis

Background

Staphylococcus aureus is an important cause of clinical mastitis in dairy cows worldwide. The cure rate after antimicrobial treatment of clinical S. aureus mastitis is very variable due to both cow and bacterial factors. Studies have shown that bacterial genotype might affect short-term bacteriological and clinical cure, but the long-term outcome has been less studied. The objectives of this study were to investigate associations between bacterial genotype and long-term outcome of veterinary-treated clinical mastitis (VTCM) caused by S. aureus during a follow-up period of 120 days and to study genotype variation among Swedish S. aureus isolates. S. aureus isolates from cases of VTCM were genotyped by pulsed-field gel electrophoresis. Long-term outcome measurements used were somatic cell count (SCC), additional diagnoses of VTCM, milk yield and culling. Isolates were classified into clusters (>80% similarity) and pulsotypes (100% similarity). Clusters and pulsotypes were grouped according to occurrence. Multivariable mixed-effect linear regression models including cow and bacterial factors with possible influence on SCC or milk yield were used to calculate differences in SCC or milk yield between groups. Additional outcome measures were calculated using a test of proportions.

Results

The isolates (n = 185) were divided into 18 clusters and 29 pulsotypes. Two pulsotypes were classified as common, and were found in 64% of the cases of VTCM. Remaining isolates were classified as less common or rare pulsotypes. The distribution was similar at cluster level. Outcome was calculated from follow-up data on 111 cows. Significantly lower SCC during the follow-up period was found in cows infected with common clusters compared to in cows infected with less common/rare clusters. The proportion of cows with SCC <200 000 cells/ml during the whole follow-up period was significantly higher in the group common clusters than in the group less common/rare clusters. Bacterial genotype did not influence the other outcome parameters.

Conclusions

In Sweden, two S. aureus pulsotypes, identified in about 64% of clinical S. aureus cases, were widespread. Cows infected with the common genotypes had significantly lower SCC during 120 days after treatment compared to cows infected with less common or rare genotypes.     

Cytotoxic Effects of Loperamide Hydrochloride on Canine Cancer Cells

Loperamide is a peripheral opiate agonist that can cause apoptosis and G2/M arrest in human cancer cell lines and may sensitize cells to chemotherapy. The objectives of this study were to investigate the effects of loperamide on viability, apoptosis and cell cycle kinetics in canine cancer cells and to establish whether the drug sensitizes cells to doxorubicin. Cell viability was assessed using Alamar Blue. Cell death and cell cycle were studied using flow cytometry with 7-Aminoactinomycin-D (7-AAD) and propidium iodide (PI), respectively. Loperamide decreased cell viability in a dose-dependent fashion and was most effective against canine osteosarcoma cells. In all cell lines, it induced a dose and time dependent apoptosis and resulted in accumulation in G0/G1. When co-incubated with doxorubicin, loperamide induced a synergistic cell kill in canine carcinoma cells. Investigation is warranted into the role of loperamide in the treatment of canine cancer.     

The Essential Role of Human Dimensions and Stakeholder Participation in States’ Fulfillment of Public Trust Responsibilities

The Public Trust Doctrine (PTD) vests states with a fiduciary responsibility to manage wildlife for the benefit of current and future generations. States have varied approaches to applying the PTD for wildlife management ranging from a traditional focus on hunters, anglers, and trappers to a progressive approach of broad inclusion of all potential stakeholders in their decision-making processes. We argue that states need to gather and incorporate more and better human dimensions (HD) information to fulfill their PTD responsibilities. We describe some of the barriers to increased use of HD and the changes in agency culture, staffing, data gathering, and decision-making processes necessary to integrate HD effectively and comport with the PTD. We conclude that in addition to increasing fulfillment of PTD responsibilities, increased use of HD information will help maintain agency relevance, increase political support, and secure broader agency funding.     

Effect of Diets Formulated with Native Peruvian Plants on Growth and Feeding Efficiency of Red Pacu (Piaractus brachypomus) Juveniles

We evaluated the effects of casein‐based semipurified diets, alone or supplemented with native Peruvian plants, on growth, feed efficiency, and histology of the digestive tract of red pacu, Piaractus brachypomus, juveniles over an 8‐wk feeding trial. Three tanks were randomly assigned to one of four casein–gelatin (40:8) diets containing a supplement of 15% wheat meal (control) or an identical level of substitution of three South American native plant as follows: camu‐camu fruit (Myrciaria dubia), aguaje fruit (Mauritia flexuosa), or maca tuber meal (Lepidium meyenii). The fish (initial weight, 2.04 ± 0.06 g) were fed experimental diets at decreasing feeding rates from 4 to 2.6% of body weight. After 8 weeks of feeding, fish fed a diet supplemented with maca meal showed significantly higher (P < 0.05) weight gain, specific growth rate, protein efficiency ratio (PER), apparent net protein utilization (NPU), and instantaneous feed intake than fish fed other diets. Feed conversion ratio (FCR), PER, and NPU in fish fed the casein–gelatin diet supplemented with maca meal were among the best ever reported in the scientific literature, 0.64 ± 0.03, 3.13 ± 0.15 and 23.8 ± 2.0, respectively. The camu‐camu meal had a negative impact on diet palatability and utilization, which resulted in slower growth. The stomach, intestine, pancreas, and pyloric caeca at the start and end of the experiment showed normal differentiation and appearance of cells and tissues. The liver parenchyma showed lipid infiltration and pigment accumulation in all samples at the initiation of the experiment and may be attributed to the period of decreased feed intake prior to the study. At the end of the study, similar histopathologies were recorded in all samples from the control and camu‐camu groups. Normal liver histology (polyhedral hepatocytes with centrally located nuclei) was observed in two of three samples from the maca group and all the samples from the group that was fed the aguaje‐supplemented diet.     

Nutritive value of partially dehulled and extruded sunflower meal for post-smolt Atlantic salmon (Salmo salar L.) in sea water

This study determined the digestibility of protein in partially dehulled sunflower meal (SFM) and then, as the main goal, the nutritive value of high‐temperature extruded (≤149°C) partially dehulled SFM (SFM_EX) for post‐smolt Atlantic salmon Salmo salar in sea water. The digestibility study was conducted using the settling column approach (‘Guelph system’) for faeces collection as described by Hajen, Higgs, Beames and Dosanjh. In the nutritive value study, triplicate groups of 50 salmon (mean weight ～116 g) in 4000‐L outdoor fibreglass tanks supplied with 25–40 L min^?1, filtered, oxygenated (dissolved oxygen, 7.0–8.5 mg L^?1), 11–12°C sea water (salinity, 29–31 g L^?1), were fed twice daily to satiation one of five steam‐pelleted dry diets that contained 422 g of digestible protein (DP) kg^?1 and ～16.4 MJ of digestible energy (DE) kg^?1 on a dry weight basis for 84 days. Low‐temperature‐dried anchovy meal (LT‐AM) comprised 68.2% of the basal diet protein whereas in four test diets, SFM_EX progressively replaced up to 33.0% of the DP provided by LT‐AM in the basal diet (SFM_EX≤271 g kg^?1 of dry matter). Sunflower meal had 87.9% DP. Diet treatment did not significantly affect specific growth rate (1.39–1.45% day^?1), feed efficiency (1.19–1.26), percentage of dietary protein retained (45.8–47.5), gross energy utilization (46.5–49.4%), per cent survival (96.0–99.3) or terminal whole body and muscle proximate compositions. We conclude that SFM_EX can comprise ≥271 g kg^?1 of the dry diet or ≥22.7% of the digestible dietary protein of post‐smolt Atlantic salmon in seawater without any adverse effects on their performance.     

Effects of modulatory agents on neurally-mediated responses of trout intestinal smooth musclein vitro

Mediators and mechanisms responsible for the inhibitory modulation of trout intestinal smooth muscle were examined using a series of putative mediators and substances known to modulate neurotransmission in mammalian systems. Frequency response relationships to transmural stimulation and concentration response relationships to 5-hydroxytryptamine, carbachol, and substance P were established on paired segments of rainbow trout intestinein vitro in the presence and absence of putative modulatory agents. Modulation of neurally-mediated contractions of trout intestine was achieved with dibutyryl cyclic AMP and forskolin, agents that increase intracellular levels of cyclic AMP. The effect appears to be at the level of the smooth muscle, since the adenylate cyclase activator, forskolin, inhibited muscarinic and serotoninergic contractions as well as transmurally stimulated contractions. Substance P-induced contractions were unaffected by forskolin. The endogenous agonists/neurotransmitters which would increase cyclic AMP levels in rainbow trout intestinal smooth muscle are as yet unknown. The effects do not appear to be modulated by vasoactive intestinal peptide (VIP), calcitonin, calcitonin gene-related peptide (CGRP), or agents that activate -adrenoceptors. Prostaglandin E₂ (PGE₂) and ₂-adrenergenic agonists are possible agents which will decrease contractility of the smooth muscle. They were only active in the proximal intestine and on transmurally stimulated contractions. The effects of both PGE₂ and ₂-agonists appear to be prejunctional, decreasing release of contractile neurotransmitters in the enteric nervous system.     

Micromonospora: An important microbe for biomedicine and potentially for biocontrol and biofuels

Micromonospora species have long been recognized as important sources of antibiotics and also for their unusual spores. However, their involvement in plant-microbe associations is poorly understood although several studies demonstrate that Micromonospora species function in biocontrol, plant growth promotion, root ecology, and in the breakdown of plant cell wall material. Our knowledge of this generally understudied group of actinomycetes has been greatly advanced by the increasing number of reports of their associations with plants, by the deployment of DNA cloning and molecular systematics techniques, and by the recent application of whole genome sequencing. Efforts to annotate the genomes of several Micromonospora species are underway. This information will greatly augment our knowledge of these versatile microorganisms.     

Non-ideal Behavior During Complete Dissolution of Organic Immiscible Liquid: 2. Ideal Porous Media

Column experiments were conducted using ideal natural sands and stainless-steel beads to examine the complete dissolution behavior of an organic immiscible liquid. Trichloroethene was used as the representative organic liquid. The elution curves exhibited multi-step behavior, with multiple extended periods of relatively constant contaminant flux. These secondary steady-state stages occurred at concentrations several orders-of-magnitude below aqueous solubility for the well-sorted sands. In contrast, the secondary steady-state stages occurred within 1 log of aqueous solubility for the poorly sorted sand. The non-ideal behavior is hypothesized to result from constraints to hydraulic accessibility of the organic liquid to flowing water, which may be expected to be mediated by the pore-scale configuration of the flow field and the fluid phases.