Int. J. Anim. Veter. Adv.

, 3(2): 77-82, 2011 inhibiting tyrosinase activity, and that they are toxic weights. Male Wistar rats were obtained from the same to certain cancer cells (Kubo al.,1994). et laboratory and introduced into each group for mating. Toyomizuet al. (2003) reveal a unique function ofThey were left in darkness overnight overnight (Akpaffiong etal., 1986). anacardic ac'd in that, for dietary conditions enhancing body fat deposition, that is consumption of a diet high in carbohydrates, dietary anacardic acid has the potential Sneaking of vaginal: On the following day after mating, the to vagina1 smear was taken irom each female rats in each group decrease body fat deposition. and The effect of the methanol leaf extract, observed to confirm mating. The materials ured were dichloromethane, ethyl acetate and w-hexane fractions 5ml syringe, beaker, distilled water, glass slides, from Anacardium occidentale Linn was investigated in and light microscope. One ml distilled water was injected streptozotocin-induced diabetic rats. Oral administration with a syringe into the vaginal of each rat after which the of vaginal fluid was collected, smeared on the glass slides methanol extract at doses of 35,175 and 250 mg kgGl significantly reduced blood glucose levels in diabetic h and then viewed under the light microscope. The presence 3 rats of sperm cells in the vaginal fluid indicated or confirmed a after administration (Sokengal.,2007). et Further studies hwe shown the anti-bacterial, positive sperm test thus a successful mating (Akpaffiong et al., 1986). antifungal and anti-tumor activities of Cashew Apple Juice (CAJ), a processed juice from the pseudofruit of the The date of mating confirmation was marked as day cashew tree (Kubo al.,1993a, b; Kozubek al.,2001) one of gestation and ihe administration of extract started et el as well as anti-oxidant effects (MeloCavalcante al., immediately. el 2003) and anti-mutagenic activity (Santos et al., 2002; Administration of extract: Extract was administered MeloCavalcante al.,2003). et There are no reports in the literature on the effects orally to the rats by carefully inserting the syringe with of aqueous extract Anacardium occidentale of (cashew) leaf needle affix into the oral cavity of the rats. Group I (control) were given distilled water while group II, HI, on pregnancy outcome of wistar lats. The Present study and therefore was undertaker, tc evaluate the effect of this IV (experimental groups) were given the extract 300 rng/kg at day 1-7 (first trimester), day 7-14 (second plant on pregnancy outcome of wistar rats. trimester) and day 14-21 (third trimester), respectively. Throughout the period of administration, food and water MATERIALS AND METHODS were given to the rats. The weight of each pregnant rat for both Plait Material: Samples of the leaves of Cashew were control and experimental groups were measured at days collected within the main campus of Ahmadu Bello 1, 8, and IS of the gestation period. Cage side examination were performed daily to detect overt signs of University in Octooer, 2010. The plant was identified toxicity (salivation, lacrimation, chewing jaw movements, authenticated by M. Musaof the herbarium section in the ptosls, Department of Biological Science, Ahmadu Bello squinting, writhing, convulsion, tremors, yellowing of fur, loss of hair, stress, erection of fur, vocalization and University, Zaira. cxophthalmia, behavioural abnormalities and moribound or Preparation of the extract: Fresh leaves of Cashew were dead rats) (Ratnasooriya et al., 2003). collected, air dried at room temperature and pulverized Parturition: After parturition, the numbers of viable or with pestle and mortar. 200 g of the powder was still macerated with 150 mL of distilled water. The extract born pups were recorded. All pups were evaluated for obtained was concentrated and evaporated to dryness ongross external congenital abnormalities (open eyelids, tail a water bsth at a temperature of 60C to obtain a brownish anomalies, club foot, oligodactyl or syndactyl). The birth mass weight, crown-rump length and head circumference oi the of 20 g. pups were determined. The birth weight of the pups was measured using Electronic Top Loading (kp -series Experimental animals: Twenty Adult Wistar rats ofboth weighning balance, obtained from department of sexes weighing 180-300 g were used for the experiment. pharmaceutics, faculty of pharmaceutical sciences A.B.U They were kept under well ventilated condition, fed on standardfeed (Excel feeds PLC) and allowed water Samaru, Zaria. The crown-rump length and head ad circumference were measures using ruler and string. libitum. Pups mortality up to 6 days, the days of eye opening and appearance of fur were observed. Grouping and mating of animals: The weight of the (Ratnasooriya et al., 2003). Based on these data, the female Wistar rats were measured using an electronic following indices were computed: weighing balance called Soehle, obtained from 5 department of Human Anatomy, Faculty of medicine Q uam al pregnancy No. of pregnant dam s X 100 No. M ated A.B.U. Zaria.The rats were grouped into four (4) groups (I, II, III and IV) of 5 rats each, according to their
78

Int. J. Anim. Veter. Adv., 3(2): 77-82, 2011

T able 1 : C hange in bod y w eight of pregnant re's on days 1 . 8 and IS of the gestation p eriod. G roupII (expt) N == 5 G e station da ys G roup! G roupIII GroupIV (Control)N-4 (expt) N - 5 (exuON-4 D a y D ay8DavlS 1 183.5*17.21?. 194.5*4.773 226.4*9.086 2328*11.236 21.8*26.852 229.2*31.879 211.5*15.521 . 231 1%.514.660 244.0*10.450 230.4*33.772 5*10.595 239.5*16.008

ii) Life birth index = No. of liveborn pureX 100 Total no. of pups bo.u iii) Gestation length iv) Litter size Statistical analysis: Data are reported as Mean Standard Error of Mean (SEM). One way analysis of variance (ANOVA) was to be statistically used to compare the weight of pregnant Wistar rats in various groups on day 1, 8 and 15 of gestation period. Birth weight, head circumference and crown-rump length of litters in control and experimental groups were checked for significance difference using student t-test. p<0.05 was deemed statistically significant. Sigma&tat 2.0 (Systat Inc, Point Richmond, CA) was used for the statistical analysis. R ESULTS

G roup III: (exp erim ental), m ean bod y w eight of rats on 1", 8* and 15* days of gestation period are 210.826.852, 229.231.879 and 230.433.772, respectively. Group IV: (experimental), mean body weight of rats on 1", 8* and 15* days of gestation period are 211.515.521, 231.510.595 and 239.516.008, respectively. In all the groups, both control and experimental, there were no statistically significant difference. Data concerning the changes in body weight of pregnant rats are presented in Table 1. Gestation period: Gestation periods of the experimental rats were observed longer than the control which is normally 21 days.

In group II, average gestation period of the two rats that Physical and behavioral observations: From littered was 24 days. observations made, there was no difference in behavioral In group III, gestation period was 22 days. changes noticed between the control group (I) and In group IV, gestation period was 25 days. experimental groups (II, and IV). There was no III Gross morphometric observation on the pups born: mortality, treatment-related sign:, of maternal toxicity, There was no still-births and/or birth abnormalities stress or abnormal behavioral changes observed within observed in all the pups bom. Some of the parameters the experimental groups throughout the gestation period. determined on the pups bom in the control and None of the pregnant rats showed vaginal bleeding or experimental group shows variations. These are: expulsion of products of conception. Overt signs of toxicity such as rhinorrhea, lacrimation, squinting and, Body weight of Pups born: The mean body weight of pups exophthalmia etc. were not expressed by the rats born in group I is 7.2200.030, group II is 5.7530.166, throughout the gestation period (Ratnasooriya et a/., group III is 5.4310.149 and group IV is 7.600C.OO. 2003). Thus, the above observation shows that the administered extract was non-toxic at the administered Litter size: The litter size of group I was 2, while that of dose. experimental groups, II, III and IV are 8, 8, 1, respectively. Body weight: The body weight (grams) of the pregnant rats in control and experimental groups, measured on the Crum-rump length: The mean Crum-rump length (cm) of first, eighth and fifteenth day of pregnancy (gestation pups born in group I is 6.450.05 while that of the period) shows variations. The mean body weight of the experimental groups; II, III and IV are 5.8750.139, rats in control: 5.7130.0639 and 6.500.00, respectively.
M th Group I: on the 1 , 8* and 15 days of gestation period are Head circumference: The mean head circumference (cm) 183.517.212, 194.514.773 and 196.5iH.660, of group I (control) is 1.1500.05, while that of the respectively. experimental groups; 11, III and IV are 1.2250.016, 1.1380.026 and 1.300.00, respectively. Group II: (experimental), mean body weight of rats1", on For all these parameters (Weight of pups born, Crum8"1 and 15* days of gestation period are 226.49.086, rump length and Head circumference), there is a 232.811.236 and 244.010.450, respectively. statistically significant differences in the mean values of the treated groups. 79

Jfit. J. Anim. Veter. Adv.. 3(2): 77-82. 2011

T able 2 : E ffects of aqueous ex tract ofrd ium o ccid ental? on soue litter raram eters A n aca leaves G roup (C ontroH n = 5 G roupII I (expt) n = 5 G roupIII (exuO n - 5 P ara m e ters M ean b irth w eight (g) M ean C row n7.2200.030 6.450*0.050 5.753*0.166 5.875*0.139 rum p length (cm ) M ean H ead 1.150*0.05 0 0 7 2 21 1 5 1.225*0.016 circum ference (cm ) Life birth index 'CO 808 24 (% ) Q uanta! p reg nancy (% ) L itter size G estation leng th (days) G ruupFV pt) n - 5 (ex

7.60*0.00 6.50*0.00 5.431*0.149 5.713*0.0639 1 1 1.138*0.026 00 8 8 22 1.30*0.00 00 7 5 25 1 0

using succinate as a substrate and hence dietary anacardic acid revealed to have the potential to decrease body fat deposition (Toyomizual.,2003). et The pups bom to rats/dams treated with leaf extract of A. ocddentale low mean birth weight and crownhas less Life birth index: All the groups are 100% Quanta pregnancy: group 3 and IV are 75% each and, rump length in group II and III compared to group I (control). However mean birth weight and mean crown-rump group II and III are 80% each. that of v) Litter size in groups I, II, III and IV are 2,8,8, and 1, length of the pups in group IV is greater than group I (control). Low mean birth weight and crown-rump length respectively, Gestation length for groups I, III, vi) I I revealed in group II and II could beresult of the action as a and IV are 21, of saponins or alkaloids and anacardic component of acid 24, 22 and respectively. 25, the extract causing impaired glucose supply to the fetuses Data concerning litters parameters are presented in and leading to Intrauterine Growth Retardation (IUGR). This is evident from the report of Pond (1996) that some Table 2. antidiabetic drugs induce IUGR. Since there was no any birth anomalies observed, low DISCUSSION birth weight observed here may be due to impaired glucose Results obtained from weighing the pregnant rats supply to the fetuses and probably not as a result or effect both in control and experimental groups on the 1", 8* and 15* day of intrinsic fetal factor such as chromosomal abnormalities of pregnancy were suojectedto statistical analysis or other malformation. to determine the mean values for each group. The Higher mean birth weight, m ean crown-rump length to differences in the mean values of the weight of treated and head circumference observed in group IV compared rats control group might have resulted due to inhibition of were not great enough to exclude the possibility that the saponins, alkaloids and anacardic acid by hormones or' difference is due to random sampling variability. Hence enzymatic action occurring during this period. It could also there is not a statistically difference in all the groups. This beadue to low litter size (one) that enhance the increase weight could have resulted from the action of anacardic acid, component of the extracx which was found to reduce of the litter. the It is important to note that length and birth weight as deposition of fat by its uncoupling (Toyomizu al., action et reported by Eide et al, (2005), each contribute 2003). Considering the pups born, with low birth weight,independently both to adult stature and body weight, even it has been established that extractA. occidentakhas though birth weight influences adult weight In past of decades, growing acceptances of the "fetal origin" of adult antidiabetic action resulting in alleviation of altered disease hypothesis have been witnessed, which has now led metabolic status in animals. However, the action by which to the sxtract lowered blood glucose in not well known, itthe thought that poor intrauterine growth is associated may increase glycogen level in liver by an increasewith risk of cardiovascular disease and Non Insulin in glycogenesis and/or a decrease in Glycogenolysis (Tedong Dependent Diabetes M ellitus (NIDDM ) in adult life. Birth weight studies have also shown that birth weight is an et al.,2006). Phytochemical analysis had revealed the presence important predictor of adult height with habies of Normal of alkaloids, polyphenols. saponins etc. and based on Birth W eight (NBW ) growing taller than babies with low the birth increased number of reports on blood glucose retardation weight, Since this present studies revealed the low birth weight associated with some saponins (Dietewa 2004) and et al., and low crown-rump length effect of A. ocddentale extract, alkaloids (Boikent al,, 2000) isolated from other et it is medicinal plants, it is liksly that the active principles therefore means that consumption of the leaf extract et al. could be present in one or the two families of the chemical during pregnancy have serious implications. Han (2000) reported that low weight has a major influence birth substances (Tedong etal., 2006). Also anacardic acid has on neurocognitive deficiencies, been shown to have an uncoupling effect on oxidative neonatal morbidity, neurobehavioural effects and phosphorylation in the rat liver mitochondria 80 For Weight of pups bom p = O.001 Crum-rump length p = 0.022 Head circumference p = 0.037

Ira. J. Anim. Veter. Adv., 3(2): 77-82, 2011 Farnsworth, N.R., 1989. Screening Plants for NewMedicine. In: Wilson, E.O. (Ed.), Biodiversity. Partll. National Academy Press, Washington, pp: 8397. CONCLUSION Han, S., D.H. Pfizeranair, E. Gracia, M.L. Eguez, M. Ling, W. Kemp and J.D. Bogden, 2000. Effects of This investigation was carried out to see the effect of A. lead exposure before pregnancy and dietry calcium occidentaleon pregnancy outcome of wistar rats. It was during pregnancy on fetal development and lead observed that there was no significant increase in accumulation. Environ. Health Perspect, 527108: bodyweight of pregnant rats during gestation period. Pups 531. born in group II and (experimental) has low mean III birth Kubo, I., I. Kinst-Hori and Y. Yokokawa, 1994. weight and less mean crown-rump length compared Tyrosinase inhibitors from to Anucardium occidentale group-1 (control), while that of Group IV control is higherfruits. J. Nat. Prod., 57: 545-51. than control. In spite of the beneficial effects of Kubo, I., M. Ochi, P.C. Vieira and S. Komatsu, 1993a. A. occidentale treatment of diseases, its adverse effect in that Antitumor agents from the cashew (Anacardium goes alongside cannot be overrule and must therefore occidentale) be apple juice. J. Agric. Food Chem., 41: taken care of. Thus from the observations made in this 1012-1015. studies, I want to conclude that the extact of Kubo, I, H. Muroi and M. Himejima, 1993b. StrutureA. occidentale should not be taken by pregnant women, even antibacterial activity relations of anacardic acids. J. when associated with diabetes, during pregnancy. Agric. Food Chem., 41: 1016-1019. Kozubek, A., R. Zarnowski, M. Stasiuk J. and ACKNOWLEDGM ENT Gubernator, 2001. Natural amphiphilic phenols as bioactive compounds. Cell. Mol. Biol. Lett., 6: 351The authors of this study wish to acknowledge the 355. technical assistance of the laboratory staff of Melocavalcante, A.A., G. Rubensam, J.N. Picada, E.G. the Department of Anatomy, Faculty of Medicine Ahmadu Silva, F.J.C. Moreira and JAJ>. Henriques, 2003. Bello University, Zaria, Nigeria. Mutagenic evaluation, antioxidant potential and antimutagenic activity against hydrogen peroxide of REFERENCES cashew (Anacardium occidentale) apple juice Tnd cajuir.a. Environ. Mol. Mutagen., 41: 360-369. Akpaffiong, MJ., GJ. Ekandem and S.P. Singh, 1986. Morton, J.F., 1987. Fruits of Warm Climates. Flair Effects of pyrimethaniine en growth and Palate Books, Florida, ISBN: 0-9610184-1-0. formation in vistar rats. West Afr.J.Anat., 1(1): 33Pond, S.M., 1996. Medicine in Pregnancy. 1st Edn., 35. Bolkent, S., R. Yanardag, Tabakoglu-ognz and O. Ozsoy- Australian Government Publishing Service. Canbeira, Australia. sacan, 2000. Effects of chard vulgatejL. var. (ft: Ratnasooriya, W.D., J.R.A.C. Jayakody and G.A.S. cicld) extract on pancreatic p cells in streptozotocin Premakumara, 2003. Adverse pregnancy outcome -diabetic rats: A morphologic and biochemical study. J. in rats following exposure to a Salacia reticulate Ethnopharmacol., 73: 231-259. Dietewa, M., B.C. Samba, H.C.T. Assah and A.A. Abena, (Celastraceae) root extract. Braz J. Med. Biol. Res., 36:931-935. 2004. Hypoglycemic and antihyperglycemic effects of Ravelli, A.CJ., J .H.P. Vander Meulen, RJ> J. Micheis, C. diethyl ether fraction isolated from the aqueous Osmond, D.J.P. Barker, C.N. Hales and O.P. extract of the leaves of Cogniauxia podoieana Baillon in normal and alloxan-induced diabetic rats. J. Bleker, 1998. Glucose tolerance in adults after prenatal exposure to famine. Lancet, 351:173-177. Ethnopharmacol., 92:229-232. Santos, M.C., J. A.P. Henriques and B. V.A. Aragoa, 2002. Eide, M.G., N. Oyen, R. Bkjaerven, S.T. Nilsen, T. Bjerkedal and G.S. Tell, 2005. Size at birth and Activity antimutagenoca critricas fruit consumed in gestational age as predictors of adult height and the human diet Stud. Biol., 24:27-32. Sokeng, S.D., D. Lontsi, P.F. Moundipa, H.B. Jatsa, P. weight. Epidemiology, 16: 175-181. Eisner, T., 1990. Chemical Prospecting: A Call for Watcho and P. Kamtchouing, 2007. Action. In: Borman, F.E. and S.R. Kellert (Eds.), Hypogiycemic effect of Anacardium occidentale L. Ecology, Economic and Ethics: The Broken Circles. methanol extract and fractions on streptozotocininduced diabetic rats. Global J. Pharmacol., 1(1): Yale University Press, New Haven, CT. 01-05.
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mortality. Ravelli el al. (1998) also reported that reduced growth in utero to be linked to decrease glucose tolerance in adult life.

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T edong, L ., T . D im o, P.D .D . D zeufiet, A .E . A songalem ,

(A nacardiaceae) leaves in streptozotocin induced A ssisted in the review of the paper, S. A bba and I. diabetic rats. A fr. J. Trad. CA M , 3(1): 23-35. Ezekiel: Colleagues in laooratory w ork. Toyom izu, M ., K. O kam oto, T. Ishibash, T. N akatsu and Y. Akiba, 2003. Reducing effect of dietary anacardic acid on body fat pads in rats. A ni. Sci. J., 74(6): 499-504.

D.S. Sokeng, P. Callard, J.F. Flejou and P. Kamtchouing, 2006. Antihyperglyccmic and renal Hamman W.O.: Major supervisor, S. Musa: Minor protective activities of Anacardium occidentale supervisor; A.D.T. Goji: Analysis of data; A.A.OyewaIe:

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