Arthur J. Goven

A non-invasive technique for sequential collection of earthworm (Lumbricus terrestris) leukocytes during subchronic immunotoxicity studies:

We have compared two methods for collecting earthworm leukocytes (coelomocytes) with respect to cell yield, viability and behaviour in immunoassays. Non-invasive extrusion was more efficient than puncturing the coelomic cavity. Extrusion does not produce trauma to earthworms maintained under long term laboratory conditions. Neither technique modified immune assays, as determined by erythrocyte and secretory rosette formation, and phagocytosis, since all 3 were functionally equivalent. After an initial extrusion, sequential leukocyte collections by extrusion are possible at intervals of 6 weeks without affecting total and differential cell counts and rosette formation.

Effect of immune system imagery on secretory IgA.

This study was an investigation of the effects of physiologically-oriented mental imagery on immune functioning. College students with normal medical histories were randomly selected to one of three groups. Subjects in Group 1 participated in short educational training on the production of secretory immunoglobulin A. They were then tested on salivary IgA, skin temperature, and the Profile of Mood States (POMS) before and after listening to a 17-minute tape of imagery instructions with specially composed background “entrainment” music designed to enhance imagery. Subjects in Group 2 (placebo controls) listened to the same music but received nor formal training on the immune system. Group 3 acted as a control and subjects were tested before and after 17 minutes of no activity. Treatment groups listened to their tapes at home on a bi-daily basis for six weeks All groups were again tested at Weeks 3 and 6. Secretory IgA was analyzed using standard radial immunodiffusion techniques. Repeated measures analyses of variance with planned orthogonal contrasts were used to evaluate the data. Significant overall increases (p<0.05) were found between pre- and posttests for all three trials. Groups 1 and 2 combined (treatment groups) yielded significantly greater increases in sIgA over Group 3 (control) for all three trials. Group 1 (imagery) was significantly higher than Group 2 (music) in antibody production for Trials 2 and 3. Symptomatology, recorded by subjects at Weeks 3 and 6, was significantly lower for three symptoms (rapid heartbeat, breathing difficulty, and jaw clenching), favoring both treatment groups over the control group.

Comparative toxicity of polychlorinated biphenyls to earthworms Eisenia foetida and Lumbricus terrestris

Effects of polychlorinated biphenyls (PCB; Aroclor 1254) in the manure worm, Eisenia foetida, on survival (LC50/LD50), and ability of coelomic leukocytes (also called coelomocytes) to form secretory rosettes (SR) and erythrocyte rosettes (ER) with, and to phagocytose antigenic rabbit red blood cells were determined and compared with those published for the earthworm, Lumbricus terrestris. Using a 5-day filter paper contact exposure protocol, LC50 and LD50 were 30.4 microg cm(-2) and 4500 microg g(-1) dry mass, respectively. Nominal PCB exposure concentrations of 5.0 and 10.0 microg cm(-2) resulted in tissue levels of 1400 and 2900 microg g(-1) dry mass. These body burdens resulted in significant reduction in SR formation by 18 and 52%, respectively. ER formation and phagocytosis were reduced 52 and 61%, respectively, only at the higher tissue concentration. Compared to L. terrestris, E. foetida: (1) accumulated considerably more PCB at each exposure concentration; (2) showed lower LC50, but higher LD50; and (3) exhibited effects on coelmocytes only at tissue PCB concentrations that caused some mortality. In terms of lethality and immunomodulation of SR, ER and phagocytosis, E. foetida appears to be more resistant to PCB than L. terrestris.

Sperm count in earthworms (Lumbricus terrestris) as a biomarker for environmental toxicology: Effects of cadmium and chlordane

Earthworms, Lumbricus terrestris, exposed in artificial soil to sublethal concentrations of technical chlordane (6.25, 12.5, and 25 ppm) and cadmium nitrate (100, 200, and 300 ppm) exhibited significant reduction in spermatozoa from testes and seminal vesicles. The onset time of reduction varied with exposure concentration, but absolute depression in sperm count was independent of exposure concentration or exposure duration after reduction was first manifested, demonstrating a threshold effect. Earthworm sperm counts show potential as a rapid-measurement endpoint biomarker for measuring sublethal effects of chemical pollutants on reproduction.

Biofeedback-assisted relaxation: Effects on phagocytic capacity

The purpose of this study was to investigate whether subjects who self-report high levels of stress have lower immunity, and whether “low”-immunity subjects under “high” stress could enhance phagocytic activity through biofeedback-assisted relaxation (BAR). During Phase 1, the level of stress and the level of phagocytic immune functioning (nitroblue tetrazolium test) were assessed as “high” or “low.” Significant chi-square analysis (x2=3.8624, df=1, p<.05) showed that subjects with “high” stress had “low” immunity. Sixteen “high”-stress, “low”-immunity subjects were randomly assigned to BAR and control groups during Phase 2. Following treatment, NBT changes showed significant increases (F=11.11, p<.003) for experimental group as compared to control group. White blood cell count and white blood cell differential were unchanged across blood samples for both groups. Experimental subjects reported significant decreases in tension-anxiety and increases in overall coping. BAR was concluded to have improved coping skills and phagocytic capacity. BAR affected the quality, rather than the quantity, of phagocytic neutrophils.

Spirituality and Cardiovascular Risk

This study extended current research linking spirituality to health by investigating the relationship between extrinsic and intrinsic spirituality and cardiovascular risk factors. Participants included 111 healthy males and females, ages 28 to 63. Measurements consisted of the Lifestyle Assessment Questionnaire (LAQ), a version of Kellys Repertory Grid, hematological analysis, and blood pressure. A greater sense of spirituality was associated with lower cholesterol risk ratios (total cholesterol/HDL) and triglyceride levels. In addition, the structural or organizational characteristics of intrinsic spirituality were associated with several other hematological measures. Further research will help clarify the association between mechanisms underlying spirituality and health, including susceptibility to cardiovascular disorders.

Earthworm Immunoassays for Evaluating Biological Effects of Exposure to Hazardous Materials

A noncontroversial and cost-effective system of laboratory and in situ bioassays capable of integrating variables of environmental concentration, route of exposure and bioavailability with a broadly applicable suite of toxic endpoints is needed to assess biological risks of environmental pollutants from hazardous and Superfund waste sites, both before and after clean-up. The system also would be useful in screening or categorizing wastes, such as industrial and municipal solids, combustion residues from incinerated solids, sewage treatment sludge, and dredged sediments for appropriate landfill disposal i.e., sanitary versus hazardous). An extensive literature on the basic biology and ecology of earthworms (Edwards and Lofty, 1977; Satchell and Martin, 1981; Satchell, 1983; Lee, 1985; Fitzpatrick et al., 1989) and from laboratory and in situ toxicity and/or bioaccumulation studies (Appendix 1, No. 1) supports using several earthworm species to develop standardized protocols (Appendix 1, No. 2) for evaluating biological risks of terrestrial pollutants.

Thermal acclimation, preference and effects on .Vo2 in the earthworm Lumbricus terrestris

Abstract 1. 1. Log .VO 2 vs log body weight yielded significant slope ( b = −0.40) and intercept ( a = 1.82) in Lumbricus terrestris acclimated and measured at 10°C. 2. 2. Temperature significantly affected .VO 2 in worms acclimated and measured at 5, 10, 15 and 20°C. 3. 3. Non-significant .VO 2 difference between 10 and 15°C (37.8 vs 43.4μlg −1 h −1 ; Q 10 = 1.3) indicates nearly complete metabolic compensation/acclimation in that range. 4. 4. Partial compensation between 5 and 10°C is indicated by Q 10 =1.8 (28.1 vs 37.8 μ1 O 2 g −1 h −1 ). 5. 5. Acclimation to 10 and 20°C showed no significant effect on thermal preference (16.3 vs 15.6°C).

Gender differences, anger expression, and cardiovascular risk.

Research has established the influence of serum cholesterol and anger management style as cardiovascular risk factors. However, mixed and sometimes contradictory results have created uncertainty with respect to contributions of anger. Some incongruities may be accounted for on the basis of gender differences since a number of studies have employed male samples rather than female samples or mixed samples. The present study examined both gender and anger management style in relation to cardiovascular risk. One hundred three (72 men and 31 women) midlevel executives completed questionnaires and a blood draw during a week long, succession training program within a large international organization. Gender, anger expression, and anger expressed outward were significant predictors for levels of high-density lipids, low-density lipids/very low-density lipids, and triglycerides, and cardiovascular risk ratios. Results replicate and extend previous research. Anger held inward and anger control were significant for men but not women, contrary to earlier gender-specific findings.

Tissue eosinophil numbers and phospholipase B activity in mice infected with Trichinella spiralis

Abstract Wilkes S. D. and Goven A. J. 1984. Tissue eosinophil numbers and phospholipase B activity in mice infected with Trichinella spiralis. International Journal for Parasitology 14. 479–482. Tissue eosinophils were counted and phospholipase B activity was assayed in the intestines of mice infected with 200 Trichinella spiralis larvae. The numbers of intestinal eosinophils and phospholipase B activity increased, peaked and returned to normal levels during the same time period. The findings support the hypothesis that a parasite-induced tissue eosinophilia is the source of elevated phospholipase B activity present in parasitized tissues.