Recovery of microorganisms from synovial and pleural fluids of animals using hyperosmolar media

L-phase (CWD) broth and plate media were used in parallel with conventional microbiological media during a 3-year period for culturing synovial and pleural fluids of animals. Two kinds of recoveries were obtained where parallel conventional methods were negative: (1) parent or normal bacteria, in very low numbers; and (2) Type B CWD variants in equally low numbers. Organisms in group 1 were: Streptococcus zooepidemicus from horses (2X); β-hemolytic streptococci, Lancefield Gp. G (2X); Staphylococcus aureus; Actinobacillus, and Actinomyces viscosus. Group 2 consisted of Bacteroides sp., Propionibacterium acnes, and three “Nocardia-like” sp. Catalase + Actinomyces was not recovered equally well on CWD plates as on conventional media with fluids obtained during ampicillin treatment. This occurred in spite of the fact that the CWD media was shown to support growth and reversion of laboratory induced L-phase variants of Nocardia caviae and N. asteroides, and had facilitated recovery of a Bacteroides L-phase variant from a pleural fluid. The nature of this fault in the media is under investigation in this laboratory.     

Evaluating protection against Loma salmonae generated from primary exposure of rainbow trout, Oncorhynchus mykiss (Walbaum), outside of the xenoma-expression temperature boundaries

A series of challenge and re-challenge studies was conducted in which juvenile rainbow trout were exposed to the pathogen Loma salmonae , a microsporidian which typically causes xenoma formation during sporogony and inflammation in the gills as the xenomas undergo dissolution. The specific goal was to determine if a primary exposure, conducted at a water temperature outside of the range which permits the parasite to undergo sporogony and form branchial xenomas, would stimulate a protective response in the fish to a later challenge conducted under temperature conditions optimal for the parasite. Primary challenge of fish to L. salmonae at 7 °C or 21 °C blocked or limited xenoma formation, as discussed in a previous study. However, these fish had a relative percentage protection (RPP) against a second optimized exposure which matched, or was not significantly less than, the degree of protection (100%) that developed in other groups of fish that received a primary exposure throughout the range of water temperatures which permits xenoma formation. When the primary exposure was conducted at 5 °C, the RPP against the second exposure was adversely affected and declined to 61%. These findings have application to the control of L. salmonae within aquaculture, in that it may be possible to expose hatchery stocks of susceptible salmonid species to spores of L. salmonae when hatchery water temperature is at 7 °C. At this temperature, the risks of disease stemming from this primary exposure appear minimal, since xenomas fail to form. However, the degree of protection appears promising, and may be sufficient to protect fish from spore exposure occurring at netpen marine sites where the parasite may be endemic.     

Female leadership raises aspirations and educational attainment for girls: a policy experiment in India

Exploiting a randomized natural experiment in India, we show that female leadership influences adolescent girls' career aspirations and educational attainment. A 1993 law reserved leadership positions for women in randomly selected village councils. Using 8453 surveys of adolescents aged 11 to 15 and their parents in 495 villages, we found that, relative to villages in which such positions were never reserved, the gender gap in aspirations closed by 20% in parents and 32% in adolescents in villages assigned a female leader for two election cycles. The gender gap in adolescent educational attainment was erased, and girls spent less time on household chores. We found no evidence of changes in young women's labor market opportunities, which suggests that the impact of women leaders primarily reflects a role model effect.     

Loma salmonae-associated growth rate suppression in rainbow trout, Oncorhynchus mykiss (Walbaum), occurs during early onset xenoma dissolution as determined by in situ hybridization and immunohistochemistry

Experimental infection of rainbow trout, Oncorhynchus mykiss (Walbaum), juveniles with Loma salmonae at a water temperature of 15 °C yielded detectable parasite DNA within the gills by week 2 post-exposure (PE) and detectable spore-wall antigen within developing xenomas by week 3 PE, as determined by in situ hybridization and monoclonal antibody (Mab) based immunohistochemistry, respectively. The microsporidian was most commonly located within endothelial cells of lamellar basal channels. Whereas the onset of xenoma formation appeared to be relatively synchronous, as expected from previous studies, xenoma dissolution followed an unexpected biphasic pattern with peaks at weeks 4 and 9 PE. The onset of significant growth rate suppression, at week 4 PE in exposed fish, was temporally associated with the appearance of gill lesions which, in turn, were centred about sites of premature xenoma dissolution. The latter was determined by the detection of spore-wall antigen within lesions. Co-habitant control fish began developing xenomas by week 10, indicating the infective potential of those spores released from the principal fish during early xenoma dissolution. Although infection with L. salmonae significantly affects fish growth rates, the time-course of this suppression is limited, and as an unexpected finding, growth rate recovery commences prior to the infection’s resolution.     

In vitro and in vivo antioxidant and anti-inflammatory capacities of an antioxidant-rich fruit and berry juice blend. Results of a pilot and randomized, double-blinded, placebo-controlled, crossover study

This study investigated the in vitro and in vivo antioxidant and anti-inflammatory properties of a juice blend (JB), MonaVie Active, containing a mixture of fruits and berries with known antioxidant activity, including acai, a palm fruit, as the predominant ingredient. The phytochemical antioxidants in the JB are primarily in the form of anthocyanins, predominantly cyanidin 3-rutoside, cyanidin 3-diglycoside, and cyanidin 3-glucoside. The cell-based antioxidant protection of erythrocytes (CAP-e) assay demonstrated that antioxidants in the JB penetrated and protected cells from oxidative damage ( p < 0.001), whereas polymorphonuclear cells showed reduced formation of reactive oxygen species ( p < 0.003) and reduced migration toward three different pro-inflammatory chemoattractants: fmlp ( p < 0.001), leukotriene B4 ( p < 0.05), and IL-8 ( p < 0.03). A randomized, double-blinded, placebo-controlled, crossover trial with 12 healthy subjects examined the JB's antioxidant activity in vivo. Blood samples at baseline, 1 h, and 2 h following consumption of the JB or placebo were tested for antioxidant capacity using several antioxidant assays and the TBARS assay, a measure of lipid peroxidation. A within subject comparison showed an increase in serum antioxidants at 1 h ( p < 0.03) and 2 h ( p < 0.015), as well as inhibition of lipid peroxidation at 2 h ( p < 0.01) postconsumption.     

Effect of water temperature manipulation on a thermal unit predictive model for Loma salmonae

Loma salmonae is an important microsporidian pathogen affecting the mariculture of Pacific salmon, Oncorhynchus tshawytscha (Walbaum). Clinical signs associated with infection arise when the parasite enters sporogony and forms xenoparasitic complexes (xenomas) within the gill. The present study tested a thermal unit (TU) model, which was devised to predict the timing of xenoma formation, under conditions in which water temperature changed during the course of infection. In vivo trials with juvenile trout showed that the model TU for xenoma onset= (days post exposure) (C above 7C) accurately predicted the onset time for xenomas when fish were moved from either 11 or 5C to 15C at various intervals after exposure. These findings suggest that the TU model may allow aquaculturists to predict disease onset. However, the model failed when fish were moved from 15 to 5C at intervals after exposure. This finding suggests that the temperature-constrained phase of the life-cycle of L. salmonae occurs early on after the spore is ingested by the fish.     

Subcutaneous abscesses and arthritis caused by a probable bacterial L-form in cats

Three cats in one household developed pyogenic subcutaneous abscesses and arthritis over a period of 9.5 months. Despite vigorous surgical and antibiotic treatment, the infections in each of these cats continued to spread locally and hematogenously to involve other joints and subcutaneous sites. Although the infections did not respond to modern broad-spectrum antibiotics, they were susceptible to tetracycline. In spite of a favorable response to tetracycline, all 3 cats were euthanatized. A causative agent could not be identified by microbiologic culture of tissues obtained prior to death and at necropsy, or with special tissue stains. The infection was transmitted experimentally by sc inoculation with cell-free material from one of the naturally infected cats to a specific-pathogen-free cat. A tissue extract from the experimentally infected cat was, in turn, infectious for another specific-pathogen-free cat. The experimentally induced lesion was a rapidly enlarging necrotizing and pyogenic cellulitis and panniculitis, with no demonstrable causative agent by special tissue stains or microbiologic culture. A probable bacterial L-form was visualized in affected tissues of the experimentally infected cats and propagated in special L-form broth. Like the natural disease, infection in experimentally inoculated cats was progressive in nature, but could be treated successfully with tetracycline.     

Induced resistance in rainbow trout, Oncorhynchus mykiss (Walbaum), to gill disease associated with the microsporidian gill parasite Loma salmonae

Branchial xenomas were detected by week 5 and disappeared by week 10 after naive juvenile rainbow trout, held at 14.5 °C, were fed or intubated with Loma salmonae ‐infected gill tissue preparations. Upon re‐challenge with L. salmonae , these fish were protected from disease and branchial xenomas did not develop. Branchial xenomas were never detected in naive fish held at 10 °C and exposed to L. salmonae . When these fish were re‐challenged with L. salmonae at 14.5 °C, they were also protected from the disease. Branchial xenomas also developed after naive fish, held at 14.5 °C, were injected intraperitoneally with a semipurified preparation of fresh spores, but generally did not develop after intraperitoneal injection with a preparation of spores subjected to freezing and thawing before use. Fish that had received fresh spores intraperitoneally were completely resistant to disease when re‐challenged via oral delivery of spores, whereas those that had received frozen spores were incompletely, but significantly, protected from disease compared with naive fish. We conclude that infection with L. salmonae induces strong protection towards the disease upon re‐exposure to spores, and that the protection does not depend on the completion of the parasite's life cycle, thus establishing the basis for further research on vaccine development for this disease.