Trino IB mixture at a very minimal concentration (0.2 ml) was found not only to protect rats against lethal dose of pathogenic isolates of Escherichia coli and Staphylococcus aureus but it is also capableof ameliorating the deleterious effect of very high concentration of sodium arsenite (6 mg/kg body weight) in rat and bird

Oloke, JK

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African Journal of Biotechnology Vol. 7 (14), pp. 2321-2323, 18 July, 2008     Available online at http://www.academicjournals.org/AJB ISSN 1684–5315 © 2008 Academic Journals     Short Communication  Immunomodulating  properties of Trino IB immunobooster  Oloke, J. K.  Pure and Applied Biology Department, Ladoke Akintola University of Technology, PMB 4000, Ogbomoso, Nigeria.  E-mail: jkoloke@yahoo.co.uk.  Accepted 20 June, 2008  Trino IB mixture at a very minimal concentration (0.2 ml) was found not only to protect rats against lethal dose of pathogenic isolates of Escherichia coli and Staphylococcus aureus but  it is also capable of ameliorating the deleterious effect of very high concentration of sodium arsenite (6 mg/kg body weight) in rat and bird.    Key words: Immunobooster, Escherichia coli, Staphylococcus aureus, sodium arsenite.   INTRODUCTION  Immunostimulators are drugs, which stimulate immune system by inducing activation or increasing activity of any of its components. Immnostimulators are of two catego-ries: specific immunostimulator, which provides antigenic specificity in immune response, such as vaccines or any antigen; non-specific immunostimulators which act irrespective of antigenic specificity, to augment immune response of other antigen or stimulate component of the immune system without antigenic specificity.  The use of immune modulators either as prophylaxis and/or part of a treatment regimen following exposure, may present a broad-spectrum approach to protect the public when exposed to a pathogenic challenge (Babineau et al., 1994). Immune modulators can increase the non specific components of the immune system such as the macrophage/neutrophil innate immune response, and stimulate the maturation of myeloid progenitor cells. Macrophage/neutrophil activation is largely considered the first line of defense against bacterial, fungal and some viral infections.  These cells are also necessary for the efficient presentation of antigens, and their level of activation determines the efficacy of the acquired immune system, the second line of defense leading to effective humoral (antibody) and cellular (T-cells) immune res-ponses (Abbas et al., 1994).  Immunomodulators have a wide range of uses as evident in the result of several works e.g. as anti-inflammatory in bowel diseases, antiinfectivity against bacteria, antiviral, antitumous, burn infection therapy, antidermatitis antiallergy, modulators of heat and cold stresses (Bill et al., 2003; Christopher et al., 2005; Di Luzio et al., 1979). So, in assessing the immunomodu-lating property of a test agent one has a choice to monitor after administration either for raised immunoglobulin like IgG or to assess directly for ability of the test agent to reduce challenged microbial load along with prevention of cancer in the presence of cancerous agents.  Trino IB is a natural product which several people in Nigeria often use to bring under control different immuno-compromised conditions. In this work the ability of Trino IB to prevent death in mice challenged with lethal dose of pathogenic Staphylococcus aureus and Escherichia coli along with its ability to ameliorate the cumulative effect of sodium arsenite challenge is used as a measure of its immunomodulating property. Intake of arsenic contami-nated water has been associated with hypertension, cancer, diabetes mellitus, atherosclerosis etc. (Chen et al., 1995; Lai et al., 1994; Tseng et al., 1996; Chiou et al., 1995). In addition, it has been shown that if 6 mg/kg of sodium arsenite is administered to rat for 26 days it produces deleterious effect on the vital organs (Mahitosh et al., 2003).   MATERIALS AND METHODS  Experimental animals  Male albino rats of 10 – 12 weeks old were purchased from central Animal House, College of Medicine, University of Ibadan. The rats were allowed to acclimatize for one week before the commence-ment of the study. Ten rats were used for each treatment. Nera black cocks of about 10 months having an average weight of approximately 2 kg were purchased from the Animal House of the Ladoke Akintola University of Technology, Ogbomoso. The birds were allowed to  acclimatize  for  one  week  before  use.  Ten2322         Afr. J. Biotechnol.    Table 1. Survival rate and bacterial load of rats given Trino IB with lethal doses of Escherichia coli and Staphylococcus aureus after 26 days of administration.  Microorganism  Rat E. coli and Trino IB E. coli only S. aureus and Trino IB S. aureus only Survival rate (%) 100 10 100 6 Bacterial load/g liver weight  0 4.43 x 107 0 7.17 x 107    Table 2. Amelioration by Trino IB of the effect of sodium arsenite on sperm count sperm motility and spermatids count.  Rat Neural black cock  Treatment Sperm count (x106/ml) Sperm motility (%) mean spermatids count ( x 106/ml) Normal saline only 72 86 45 Sodium arsenite only 10 6 23 Sodium arsenite and Vitamin E 71 40 20 Sodium arsenite and Trino IB 71 72 38    birds were used per treatment.    Bacteria  While S. aureus was isolated from a patient’s pus, E. coli was isolated from urine of a hospitalized patient at Baptist Medical Centre, Ogbomoso.    Rats  Using cannula, 0.2 ml (4.0 x 107/ml) bacterial suspension and/or 0.2 ml Trino IB were administered by gavage along with or without 0.2 ml of 6 mg/kg body weight of sodium arsenate to different groups of rats daily for 26 days.  Vitamin E was used as positive control.  Rats were given 0.2 mol of 0.94 mg/kg body weight of Vitamin E by ga-vage daily with or without sodium arsenate for 26 days. On the 27th day, six rats were randomly selected from each group and killed by cervical dislocation. Blood samples were collected by cardiac puncture. Heparinized plasma was prepared and taken for hormone radio-immunoo assay.  Rats’ scrotal sacks were opened immediately and the testis weighed (result not shown).  Epididymis was trimmed off the right testis; their cauda epididymis was cut off the epididymal length and placed in a beaker containing 1 ml of physiological saline solution. The cauda epididymis was then lacerated at many spots so that their spermatozoa would swim out into the solution within a few minutes. Epididymal sperm motility and concentration were deter-mined using an adaptation of Brooks and Welbstesr (1975).   The heart, liver, testis and kidneys were dissected out, trimmed the attaching tissues and weighed (result not shown). The organs were fixed in formol-saline for histological study.  Microbial clearance was determined by homogenizing livers of surviving rats in 20 ml sterile nutrient broth. Diluents were plated on nutrient agar and incubated for 24 h at 37oC. Bacterial load per liver gram weight was computed.    Birds  Using cannula, 0.2 ml of 6 mg/kg body weight of sodium arsenite with or without 0.2 ml of Trino IB mixture was administered  to each bird for 5 weeks consecutively  i.e, day 7, 14, 21, 28 and 35.  Vitamin E served as positive control. Vitamin E was administered by gavage once daily. The dose given was 6.7 mg/bird.  The birds were bled using needles and syringes to draw the blood from the main vein in the armpit. The blood was used for hormonal assay. Organs collected for histophathogy test from slaughtered bird include liver, kidney, heart, and spleen. Elongated spermatids count was carried out (Brooks and Welbstesr , 1975).    RESULT AND DISCUSSION  All the rats given lethal doses of E. coli and S. aureus along with Trino IB survived throughout the 26 days of experiment while 10 and 6% respectively of those given E. coli and S. aureus only survived (Table 1). In addition, while there were no organism in the liver of those animals that were given Trino IB with the bacteria, while heavy bacterial load was detected in the liver of those animals given bacteria only (Table 1).  When sodium arsenite alone was administered to rats and neural black cocks, it consistently has a very deleterious effect on sperm count, sperm motility, mean spermatid count and on the different hormonal systems (Tables 2, 3). Trino IB mixture however, also consistently ameliorates the deleterious effect of sodium arsenite on sperm count, sperm motility and hormonal system of the animals. From the results obtained in this work, Trino IB ameliorates the effect of sodium arsenite better than vitamin E which is a known antioxidant.   The histopathology examinations show that the spleen of the group treated with sodium arsenite alone show moderate hyperplasia of the lymph nodes with haemo-siderosis. In addition the liver of the animals treated with sodium arsenite alone also demonstrated significant haemosiderosis with a moderate testicular hypoplasia of some somniferous tubules of the testis. The groups treated with sodium arsenite along with either Trino IB or Vitamin E appeared normal.  Oloke        2323    Table 3. Amelioration by Trino IB of the effect of sodium arsenite on the hormonal status of rat and neural black cock.   Rat   Treatment Leutenizing hormone (mlU/ml) Follicle stimulating hormone (mlU/ml) Testosterone (ng/ml) Neural Black cocktestosterone (ng/ml) Normal saline only 3.75 2.25 0.85 2.5 Sodium arsenite only 2.15 1.5 0.40 0.6 Sodium arsenite and Vitamin E 2.75 1.35 0.65 1.4 Sodium arsenite and Trino IB 3.51 2.10 0.77 2.11    Conclusion  Since Trino IB mixture not only protect rats against lethal dose of pathological isolates of E. coli and S. aureus and since it is also capable of ameliorating the deleterious effect of sodium arsenite in rat and bird, the mixture should be regarded as an excellent immune modulator.   REFERENCES  Abbas AK, Litchman AH, Pober JS (1994). Cellular and Molecular Immunology 2nd Edition Philadelphia, Pa: WB Saunders Company, p. 77. Babineau TJ, Hackford A, Kenier A, Bistrian B, Forse RA (1994). A phase II multicentre, doube-blind randomized, placebo-controlled studies of three dosages of an immunomodulator (PGG glucan) in high-risk surgical patient. Arch. Surg. (11): 1204-1210. Bill K, Rosemande M, Paulinal B, Gary O (2003). Anthrax Protective effects of yeast Betal,3.glucan. Medscape  Gen. Med. 5(1): 25-30. Brooks AK, Welbster BW (1975). CRC handbook of Laboratory diagnostic and treatment of infertility. CRC Press. Boc a Raton chap 2: 37-39. Chen CJ, Hsueh YM, Lai MS (1995). Increase prevalence of hypertension and Long-term arsenic exposure. Hypertension, 25: 53-60.                              Chiou HY, Hsuah YM, Liaw KG (1995). Incidence of internal cancers and ingested inorganic Arsenic. A seven Year Follow-up study in Taiwan. Cancer Res. 55: 1296-1300. Christopher H, Metzger G, Terrina V, Martin G, Arvydas D (2005). Immunomodulators for inflammatory bowel disease. Ann. Rev. Med. 52: 299-318. Di Luzio NR, William DL, McNamee RB, Edwards BF, Kitahama A (1979). Comparative tumor inhibitory and antibacterial activity of soluble and particulate glucan. Int. J. Cancer 24: 773-779. Lai MS, Hsueh YM, Chien CJ (1994). Ingested inorganic arsenic and prevalence of diabetes millitus- Am. J. Epidemiol. 139: 484-492. Mahitosh S, Gargi RC, Aloke C, Narendra MB (2003). Effect of sodium arsenite on spermatogenesis, plasma gonadotrophins and testosterone in rats. Asian J. Androl. 5: 27-31. Tseng C, Chong C, Chen C, Tai T (1996). Dose –Response relationship between peripheral vascular disease and ingested inorganic arsenic among residents in black foot disease-endemic villages in Taiwan. Atherosclerosis 120: 125-133. 

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Immunomodulating properties of Trino IB immunobooster

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Immunomodulating properties of Trino IB immunobooster

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