Aluminium-magnesium silicate inhibits parvovirus and cures infected dogs

doi:10.4236/health.2010.210179

Paper Menu >>

Journal Menu >>

Vol.2, No.10, 1215-1217 (2010) Health

doi:10.4236/health.2010.210179

Aluminium-magnesium silicate inhibits parvovirus and

cures infected dogs

Ezeibe Maduike C. O.1*, Nwaogu Innocent C.2, Nwigwe Ada N.1, Okorafor Obianuju N.1, Eze

James I.1, Ngene Augustine A.1

1Department of Veterinary Medicine, University of Nigeria, Nsukka, Nigeria; *Corresponding Author: maduikeezeibe@yahoo.com

2Department of Veterinary Anatomy, University of Nigeria, Nsukka, Nigeria

Received 29 October 2009; revised 25 November 2009; accepted 5 December 2009.

ABSTRACT

Ability of a synthetic Aluminium-Magnesium Si-

licate [AMS] to inhibit activities of canine parvo-

virus [CPV] was investigated in vitro and in vivo.

Five samples of CPV isolated in Nigeria, were

each incubated with equal amount of a synthetic

AMS on a volume to weight [v/w] basis, for one

hour and then centrifuged. Viral titres of the

supernatants were tested by the haemaggluti-

nation [HA] test and their mean titre compared

with mean titre of portions of same viral sam-

ples, not incubated with the AMS. Also, five

puppies and five adult dogs infected with the

parvovirus isolates were treated by dosing each

with 400 mg/kg of a drug formulation that has

12% AMS per os for seven days. As control, five

puppies and five adult dogs from same class as

the experimental dogs were similarly infected

but were not treated. Incubating parvovirus with

AMS reduced its load from mean HA titre 825.6 ±

261.1 to mean HA, 270.8 ± 132.1 [p < 0.05]. Also

treating parvovirus infected dogs with a 12%

AMS drug formulation reduced mortality due to

the virus from 100% to zero [p < 0.01].

Keywords: Aluminium-Magnesium Silicate; Canine

Parvovirus Haemagglutination Test

1. INTRODUCTION

Canine parvovirus disease is a contagious disease of do-

gs and wild canids. It affects all breeds of domestic dogs,

foxes and wolves [1,2]. The disease was first recognized

in the United States of America in 1978 [3]. It has since

then been reported in many other countries including

Nigeria [4].

Causative agent of parvovirus disease is a parvovirus

[2,5]. The disease has been reported to be more severe in

puppies than in adult dogs [6]. It is said to cause gastro-

enteritis in adults [7] but myocarditis in puppies [8]. It

leads to high mortality among dogs [9]. Since emergence

of CPV in early 1980s it has posed very serious problem

to dog breeding, yet, there is no treatment for the disease

[5]. Control of the disease is mainly by vaccination and

by hygienic measures [10]. Recent reports show that

even vaccination is no longer effective in controlling

CPV disease due to constant mutation of the virus [11].

Carmichael [12] has therefore suggested that other ways

to combat the disease be looked for.

Aluminium-Magnesium Silicate is an ore which oc-

curs naturally as mineral deposits in India and in the

United States of America [13,14]. Natural AMS, when

purified, is used as medicine for both humans and for

animals [15,16]. The EEC committee on veterinary me-

dicinal products has declared it safe for use even on food

animals [17]. Vanderbilt [13] also reported that AMS is

used to bind drugs in making tablets which are used in

treating humans and animals, because its molecules have

both positively and negatively charged ends.

Since viruses also have electrical charges [18], if

AMS is used as medicine, extracellular viruses may ad-

sorb onto it and so fail to attach to host cells. This would

disrupt the first stage of viral infection which is adsorp-

tion to the host cells’ membranes. To get a pure form of

AMS, devoid of the many impurities found in the natural

AMS, aluminun silicate and magnesium silicate were

reacted under a controlled ionic condition [19].

The synthetic AMS has been reported to reduce titre

of Newcastle disease virus [NDV] in vitro and to reduce

mortality of NDV infected chicks in vivo [20]. Haem-

magglutination [HA] titre and sero-conversion abi- lity

of Egg drop syndrome 76 virus also reduced following

incubation with the synthetic AMS [21]. The synthetic

AMS has also proved to have antiviral effects against

Infectious Bursal Disease virus [22] and against Peste

des Petits Ruminants virus [23].

Ezeibe M. C. O. et al. / Health 2 (2010) 1215-1217

1216

It was therefore thought useful to test effect of the syn-

thetic AMS on canine parvovirus both in vitro and in vivo.

2. MATERIALS AND METHODS

Five samples of CPV isolated in Nigeria (National Vet-

erinary Research Institute, Vom, Nigeria) were used for

the experiments. AMS used on the viral samples was

synthesized by reacting aluminium silicate with magne-

sium silicate [19]. A portion of each of the 5 viral sam-

ples was mixed with equal amount of the AMS on a vol-

ume to weight (v/w) basis, and allowed to stand at room

temperature for one hour before they were centrifuged at

3000 revolutions per minute for 10 minutes. The viral

supernatants were then tested for CPV titres by HA test

using 0.6% porcine RBC prepared as described by Wosu

[24] as indicator, as control, intact portions of the 5 viral

samples were similarly used for HA test on the same

plates used for their supernatants. RBC controls were

included in the protocol and the setup was incubated at

4°C [24]. Mean HA titre of supernatants of the viral

samples incubated with AMS was compared with mean

titre of their portions which were not incubated with the

AMS before the HA test.

For the in vivo experiment, 20 dogs (10 puppies and

10 adults) were acclimatized for seven days during whi-

ch they were treated with antibiotics (penicillin and stre-

ptomycin), vitamin B complex and Ivermectin to reduce

bacterial and parasitic infections. Their rectal tempera-

tures were also measured daily to ensure they were not

incubating viral infections. Each of the dogs was then

dosed with 2 ml of a CPV sample which had HA titre of

4096. They were observed daily for clinical signs of ca-

nine parvovirus disease including fever, anorexia, vom-

ition and diarrhoea. At first sign of establishment of in-

fection (fever), the experimental dogs were randomly

divided into two groups each of 5 puppies and 5 adults.

One group was treated by administering each dog with a

drug containing 12% AMS in a base of dextrose mono-

hydrate and aluminium silicate, at rate of 400 mg per kg

body weight per os, for seven days. The second group

served as untreated controls.

Mortality rates of the two groups were compared by

chi-square. Also, gross pathologic and histopathologic

lesions on the stomachs, intestines, lungs, hearts and

livers of samples of the two groups were studied and

compared to assess ability of the AMS drug to inhibit

activities of the CPV in vivo.

3. RESULTS

Incubating samples of Canine parvovirus with the syn-

thetic AMS reduced their viral loaf from a mean HA titre

of 875.6 ± 261.7 to 270.8 ± 132.1 (P < 0.05). All the

dogs treated with the AMS (adults and puppies) recov-

ered while all the untreated controls died. Also, gross pa-

thology of the untreated controls revealed swollen heart

with rounded apex, pale - swollen lungs and congested

liver in the puppies. The untreated adult dogs had disco-

loured and swollen intestines, congested and swollen liv-

er and pale - swollen lungs on gross pathologic examina-

tion. Histopathologic examination revealed necrosis and

cellular infiltration of the crypts of the duodenum, ne-

crosis of hepatocytes, presence of pyknotic hepatocytes,

dilation of the hepatic central veins and necrosis of the

myocardial cells. There was no gross pathology in the

treated cases sacrificed, except for pale lungs. Regener-

ating cells were also observed in the liver and duodenum

of the treated samples at histopathologic level.

HA titres of samples of CPV incubated with the syn-

thetic AMS are as shown on Table 1.

4. DISCUSSION

Incubating CPV samples with the AMS reduced their

HA titres. This agrees with earlier results with Newcastle

Disease Virus [20], Egg drop syndrome 76 virus [21],

Infectious Bursal Disease virus [22] and with Peste des

Petits Ruminants Virus [23]. Venkatachrisnan and Che-

nicoff [25] reported that Aluminium-Magnesium Silicate

molecules posses both positive and negative electrically

charged ends. Viral genomes also have electrical charges

[18]. So the CPV particles may have adsorbed onto the

AMS by electrostatic attraction and were removed in the

in vitro experiment.

AMS is insoluble [26] and un-absorbable [13], yet re-

sults of the in vivo experiments suggest it was able to get

to the heart, lungs and liver to inhibit pathologic activi-

ties of the CPV in those organs. It may have passed

through the mucous membranes of the gastrointestinal

tract due to the gastroenteritis caused by the viral infec-

tion [27]. Murray et al. [28] also reported that simple

sugars carry charged molecules across intact mucous

membranes by active transport. The AMS may also have

been carried across even intact mucous membranes into

the blood circulation by dextrose monohydrate mole-

cules in the drug formulation.

Table 1. Haemagglutination titre of canine parvovirus vacc-

ine samples incubated with AMS.

HA Titre

CPV samples Incubated with

AMS Controls

1 2 32

2 8 512

3 68 512

4 256 1024

5 1024 2048

270.8 ± 137.07 825.6 ± 261.07

Incubating CPV with the AMS reduced its viral titre [P < 0.05]

Ezeibe M. C. O. et al. / Health 2 (2010) 1215-1217

121

1217

Suggestion that the AMS is able to reach blood circu-

lation is supported by use of AMS as haemopurifier in

women with menstrual pains [14].

This appears a successful use of AMS for treatment of

a mammalian viral disease. Windholz [15] reported that

AMS is safe when consumed by man and by animals. It

has been in use for treatment of gastric ulcer for many

years [16]. The two chemicals used to synthesize the

pure AMS used in this study, aluminium silicate and

magnesium silicate are also medicines approved for

treatment of animals and man. So, results of these ex-

periments appear to suggest that the synthetic Alumin-

ium-Magnesium Silicate may be useful in treatment of

viral diseases of animals and man.

REFERENCES

[1] Baker, I.K., Povey, R.C. and Voigt, D.R. (1983) Re-

sponse of mink, skunk, raccon and red fox to Canine

parvovirus and feline panleukopaenia virus in Ontario.

Journal of Complementary Medicine, 47, 188-197.

[2] Nigel, D., Easton, A. and Leppard, K. (2007) Introduc-

tion to morden virology. Venetia Saunders, Kindle Store.

[3] Ford, R.B. (1988) Clinical signs and diagnosis in small

animal practice. Churchill Livingstone, New York.

[4] Kamalu, B.P. (1982) Canine parvovirus infections in

Nigeria. Journal of Small Animal Practice, 26(11), 663-

668.

[5] Khan, C. and Line, S. (2005) The Merk veterinary man-

ual. 9th Edition, Merck and Co, New Jersey.

[6] Greene, C.E. (1984) Clinical microbiology and infectious

diseases of dogs and cats. W. B. Saunders, Philadelphia.

[7] Appel, M.J. and Parish, C.R. (1987) Canine Parvovirus

type-2 of carnivores. Elsevier, New York.

[8] Pollock, R.V.H. (1984) The Parvovirus PART 11, Ca-

nine Parvovirus. Comparat. Cont. Educ., 6(1), 653-664.

[9] Thompson, N., McLandish, I.A.P., Cornwell, N.J.C.

(1979) Isolation of parvovirus from dogs in Britain. Vet-

erinary Record, 105(8), 167-168.

[10] Walsh, F.M. and Ames, S.G.B. (2004) Microbiology and

Drug resistance mechanism of fully resistant pathogens.

Nature Reviews Microbiology, 56(1), 65-79.

[11] Payne, D. and Thomas, A. (2004) Aspects of the diagno-

sis, pathogenesis and epidemiology of canine parvovirus.

Australian Veterinary Journal, 60(11), 197-200.

[12] Carmichael, L. (2005) Neonatal viral infections of pups.

International Veterinary Information Service. Ithaca,

New York.

[13] Vanderbilt, R.T. (1992) Veegum the versatile ingredient

for pharmaceutical formulation. RT Vanderbilt Co., Inc.

Technical Literature, Veegum.

[14] Jacyna, M.R. and Wormsley, K.G. (2005) Aluminium-

Magnesium Silicate used in drug formulation and cos-

metic. Bulletin of Materials Science, 24(6), 377-382.

[15] Windholz, M., Budavan, S. and Fertig, M. (1978) The

Merck index. Merck and Co Inc., New Jersey.

[16] Foster, E. and Smith, K. (2007) Aluminium-Magnesium

Silicate is used as antacids, antiulcer and control of diar-

rhoeas. Journal of Renal Nutrition, 4(2), 50-53.

[17] Schils, S. (2002) The use of montmorillonite in the fight

against harmful affects of ammonia. Journal of Renal

Nutrition, 4(2), 32-36.

[18] Cann, A.J. (1993) Principles of molecular biology. Aca-

demic Press, San Diego.

[19] Ezeibe, M.C.O. (2006) Admacine®. Federal Republic of

Nigeria Patents and Designs Act. Cap 344 LFN 1990,

NO. RP 16446.

[20] Ijabo, O. (2006) Effect of synthetic aluminium magne-

sium silicate on pestes des petits ruminants and newcas-

tle disease virus. M.Sc. Thesis, University of Nigeria,

Nsukka, Nigeria.

[21] Ezeibe, M.C.O., Okoroafor, O.N., Ijabo, O., Ukomadu, N.

M., Ngene, A.A., Eze, J.I. and Orajaka, L.J.E. (2010).

Haemagglutination and haemagglutination-inhibition ti-

tre of egg drop syndrome-76 virus treated with alumin-

ium-magnesium silicate. Animal Science, 4(3), 67-70.

[22] Ezeibe, M.C.O., Mbuko, I.J., Okoroafor, O.N., Okon-

kwo, A.C., Animoke, P., Orajaka, L.J.E and Ngene, A.A.

(2009a) In vitro and in vivo effects of aluminium-mag-

nesium silicate on infectious bursal disease virus of

chicken. Anim. Sc. Rep., 3(4), 132-137.

[23] Ezeibe, M.C.O., Ijabo, O., Okoroafor, O.N., Orajaka, L.J.

E. Ukomadu, N.M., Chukwu, O.S. and Ngene, A.A.

(2009b) Antiviral effects of aluminium-magnesium sili-

cate on Peste des Petits Ruminants Virus. Anim. Sc. Rep.,

3(4), 141-147.

[24] Wosu, L.O. (1984) Standardization of red blood cells for

haemagglutination test and removal of naturel agglutin-

ins. Nigerian Veterinary Journal, 13(1), 39-42.

[25] Chernicoff, S. and Venkatakrishnan, R. (1995) Introduc-

tion to physical geology. Worth Pub, New York.

[26] Galindo, L.A. and Cereso, P. (2006) Compositional te-

chnical and safety specification of clay to be used as

pharmaceutical and cosmetic products. Journal of Renal

Nutrition, 2(1), 38-40.

[27] Greene, C. E. (1990) Infectious diseases of dogs and cats.

W. B. Saunders, Philadephia.

[28] Murray, K.R. (2000) Harpers biochemistry. McGraw Hill,

New York.