EFFECT OF ADDING NATURAL MIXTURE JUICE OF ONION; GARLIC AND LEMON(NMJ) IN FILTER DRINKING WATER ON 1- PERFORMANCE OF GROWING RABBITS.

EFFECT OF ADDING NATURAL MIXTURE JUICE OF ONION; GARLIC AND LEMON(NMJ) IN FILTER DRINKING WATER ON 1- PERFORMANCE OF GROWING RABBITS.

Azza M. M. Badr^*; Neamat I. Bassuony^*; Fadila M.Easa^** and   M. F.S.Hanna^**

*  Regional Center for Food and Feed (Animal Nutrition),

*  Regional Center for Food and Feed (Microbiology Department),

** Animal Production Research Institute, Agriculture Research Center, Giza, Egypt

Azzabadr805@yahoo.com

A total number of two hundred twenty hybred rabbits with an initial live body weight, 751.8±22.4 g at 33 days of age, were allotted at random to four similar experimental groups (55 rabbits each). All rabbits were fed similar concentrate feed mixture (CFM) pelleted diet (16.72% crude protein and 13.07% crude fiber on average.). The first group (D¹1, control) free additive (NMJ), while groups (D², D³and D⁴)were supplemented with1, 2 and 3 ml natural mixture juice (NMJ)/ L.of garlic, onion and lemon filter drinking water, respectively. The feeding trail extended for 4 weeks. The aim of this study was to evaluate the effect of natural mixture   of garlic, onion and lemon juice as feed additive  on daily feed intake and weight gain, feed conversion ratio and efficiency, dressing percentage and internal organs weight and post weaning mortality rate of growing rabbits.

Results indicated that, daily feed intake of rabbits increased significantly (P<0.05)  by increasing NMJ in filter drinking water. Also, introducing different levels of (NMJ) significantly increased daily weight gain . Rabbits recieved 3 ml/L,  2 ml/L or 1 ml/L grew faster than control, respectively. Final body weight of rabbits supplemented with 2 and 3 ml/L (NMJ) recorded thehighest values than those supplemented with  1 ml /L (NMJ) or control diets through the 4 weeks of the experiment. There were significant (P<0.05) differences among treatments in feed conversion ratio through  the first 3 weeks ( Table 4 ). A relatively low mortility rate (10.90 and 12.73%) was observed in rabbits with  adding 3 and 2 ml/L (NMJ) in filter drinking water, while rabbits of control recorded the highest mortality rates (21.818%) during the whole experimental period. Hot carcass weight and dressing percentages were significantly (P<0.05) higher in the 1, 2 and 3 ML/L natural juice of (NMJ) groups than those of the control group. For the microbiological quality of used additive to drinking filter water,the increasing level of supplementation decreased aerobic bacterial,  coliform; feacal coliform;  enterobacteria;  yeast and fungi counts while E.coli wasnotdetected.It could be concluded that supplemention of natural mixture juice of garlic,onion and lemon(NMJ) up to 3ml /L in the filter drinking water of hybred rabbits had good results without adverse effects on growth performance.

Key words: Rabbits, Juice garlic, lemon & andonion, growth performanc, microbiological quality of the drinking water.

Feed additives   are important  manipulators that can be usedto improve the efficiency of feed utilization and animal  productivity. The world health orgnization (WHO) encourages the  acceptance of feed additives (Line-Eric et al., 1998 and Aboul-Fotouh et al., 1999). Antioxidants, antibiotics, flavouring agents, coccidiostat, antimolds and anticaking agents are synthetic feed additives used to improve the quality of feed for our livestock and their performance. However, current research is looking for natural alternatives to synthetic feed additives. Medicinal herbs such as garlic (Allium sativum), onion (Allium cepa) and limon (Citrus limon (L.) Burm) have been reported to possess antibacterial, antiseptic, anti-inflammatory, antiparasitic and immunomodulatory properties (Muhammad et al., 2009 and Thomson et al.(2007). To ensure more net return and to minimize expenditure on feed are the main challenges, for which many research strategies has been practiced such as introducing feed additives   (Pervez, 1992).

Garlic (Allium sativum) is one of the most popular herbs used world wide to reduce various risk factors associated with several diseases (Thomson et al., 2007). Actually, garlic contains a variety of effective compounds that exhibit anticoagulant (Augusti,  and Sheela,  1996), antioxidant (Anwar and  Meki 2003), antibiotic (Rees et al., 1993), hypocholesterolaemic (Ali, and Thomson1995)   hypoglycaemic, as well as, hypotensive activities (Ali, et al., 2000). Banerjee and Maulik( 2002) showed freshly crushed garlic contains allicin, alliin, ajoene, diallylsulfied, dithiin and s-allylcysteine (Onu, 2010 and Mahmoud et al., 2010) reported that the levels of 0.25, 0.50 and 1% of garlic juice in layer hens improved performance characteristics, egg quality and showed lower bacterial count of the E. coli. Garlic extract contributes to the alleviation of gastrointestinal infections in goats by reducing the coccidian burden and may enhance animal performance in adult goats (Mulumeb et al., 2009).

Garlic has been used to treat animals that suffer from gastrointestinal parasitism (Guarrera, 1999). The efficacy of garlic on coccidian infections has been reported in rabbits (Toulah and Al-Rawi, 2007). There were significant increase in life body weight and life body weight gain and feed conversion rate of Muscovy ducks which fed rations supplemented with 3% garlic and 3% garlic with 1% onion, respectively (Ibrahim et al., 2004). Ahmed et al.  (2009) recommendation is that the optimal additive level based on animal health, nutrient digestibility and daily gain was 2.5%. Thereby, natural additive of juice of lemon, onion and garlic could be used successfully and safety in rations to improve the performance of growing calves. Onions contain chemical compounds such as phenolics and flavonoids that basic research shows to have potential anti-inflammatory, hypocholesterolaemic, anticancer and antioxidant properties. Also, quercetin (Slimestad et.al, 2007) and its glycosides quercetin 3, 4’-diglucoside and quercetin-4'-glucoside(Williamson et al., 1997 and Olsson   et al., 2010) are present in onions.The citrus lemon juice (1 ml /kg /day) revealed a significant reduction in serum cholesterol, triglycerides; low density lipoprotein levels and resulted in an increase in high density lipoprotein. These results suggest that the hypocholesterolemic effects of citrus lemon juice may be due to its antioxidant effect (Yasmin, et al., 2010). The active antioxidant compounds are flavonoids, isoflavones, flavones, anthocyanins, coumarins, lignans, catechins and isocatechins. In addition, some compounds found in natural foods such as, vitamins C and E, b-carotene and a-tocopherol are  known to possess antioxidantial (Prior 2003).

On the other hand, Gupta et al. (2005) found that mixture improvement in the digestibility coefficients of different nutrients is probably due to improved gross activity of rumen microflora, increased immunity alternation in numbers and species of microorganisms in the rumen on inclusion vegetable and fruits increase in cellulolytic bacteria, increased total volatile fatty acids (TVFA's) high DM, and their TDN intake and high gain rate

Therefore, the aim of this study was to evaluate the effect of natural mixture (vegetables and fruits), garlic, and onion and lemonade juice on daily feed intake and weight gain, feed conversion ratio and efficiency, dressing percentage and internal organs weight and post- weaning mortality. Also, determinations of microbial counting drinking filter water.

MATERIALS AND METHODS

This work was carried out at privet farm belonging to IBEX company and the laboratory part of fungi, yeast count and bacteria estimation were conducted at Regional Center for Food and Feed (RCFE),  Agriculture Research Center. In this study two hundred twenty growing rabbits with an initial live body weight 751.8±22.4g at 33 day of age. The exprimental rabbits were divided randomly into four groups (55 rabbits in each). Animals were allotted randomly into four groups. The 1^st group (as control) reared on concentrate feed mixture (CFM), while the 2^nd, 3^rd and 4^th groups  supplemented with natural mixture  juice of  garlic, onion and lemon in drinking water by 1, 2 and 3 ml / L. water / day, respectively. The Juice was prepared by Neamat (2004) and modified by Mahesar et al. (2010), where fresh garlic bulbs, onion, and Limon were obtained from the local market and cut into small pieces. About 250 ml of distilled water per 100 g of peeled onion/ or garlic and Limon were added and crushed in a mixing machine in the presence of some crushed ice. The resultant slurry of every one was squeezed and filtered 3 times through a fine cloth. The aqueous extract of 3 materials was stored in dark brown bottle with scrow cap at 4 °C until use.

All rabbits reared on concentrate feed mixture (CFM). Feed ingredients and chemical analysis of the experimental diets are shown in Table 1. Fresh filter water and the natural mixture juice  were automatically available at all times through stainless steel nipples for each cage. The experimental diets were offered to rabbits adlibitum. The rations were adjusted every week according to the average daily feed intake. Rabbits were weekly weighted in the morning before offering  feed or water. Live body weight change and feed intake were recorded at weekly intervals. Feed conversion ratio was calculated as g feed/g gain. Performance index (PI%) was calculated (from  4^th  to 8^th  weeks of age) according to North (1981) as PI = Live body weight (kg) x 100 / Feed conversion.

Hot carcass weight and dressing percentage: At the end of the feeding period (60 days), 10 rabbits were fasted for 16 hrs, weighted and slaughtered. Hot carcass weight and giblets (liver, heart and kidney) as a percentage of pre-slaughter weight was determined. The dressing percentage was calculated as the hot carcass weight in addition to giblets weight (dressed weight) divided by pre-slaughter weight.

Drinking filter water microbial determination: Appropriate dilutions  prepared   from   each   sample   were  used for   inoculating  different 

Table 1:Theingredients and chemical composition of thepellet ration fed to rabbits, during the experimental period

* Calculated according to NRC (1977) for rabbits.

samples of water in selective media. The microbial determinations were applied as follows:

Total Aerobic Viable Counts: Aerobicbacterial counts were estimated on glucose yeast extract nutrient agar medium as the method reported by A.P.H.A. (2005) using pouring plate technique. Suitable plates were counted after incubation at 37°C for 48 hours.

Coliform  and   Feacal    Coliform:  Coliform  and feacal coliform counts were estimated on Mac-Conkey agar  (1905) using  pouring  plate  technique.  Suitable plates  were   counted  after 24 hours  at  37°C and 44.5°C for total coliform and feacal coliform  counts, respectively.

Entero-bacteriacea Counts:Violetred bile glucose agar medium plates (European Directorate for the Quality of Medicines and Healthcare, 2008) were incubated with 1 ml of the appropriate dilutions  andincubated overnight at37°C. After incubatation on clearly visible purple colonies surrounded by a purple halo were estimated  as entero bacteriaceae counts (Difco, 2000).

Detection of Salmonella:The methods of Goorgola and Boothroyd (1965) and Khan and Mcaskey (1973)was applied by adding 225ml peptone water as preen richment medium to twenty-five g. of each sample and incubated at 37°C for 24 hours. After incubation the culture  was streaked on difco brilliant green agar plates and examined after 25-28 hours (on this medium presumptive salmonella appears as pink colonies surrounded by brightred medium).

Enumeration of Escherichia coli 0157: H7: Culturemedia   and imurogenetic separation reagents, the enrichment medium was modified tryptone soya  broth (MTSB=N) containing novobiocin solution 20 mg/liter of  MTSB  (Bolton, et al. 1995 and ISO 6887, 2001) and the subculture on medium sorbitol Mac-Conkey agar (1905) containing defixime 1ml/ liter and potassium telluride1 ml/ liter of sorbitol Mac- Conkey agar (Cefixime etlluritesorbitol Mac-Conkey agar (CT-SMAC) (Zadik et al., 1994).

Total counts of yeast: were determined in a ruse-bengle chloramphenicol agar according the methods described in (Oxpoid Manual 2000). Plates were incubated at 22-25°C for 7 days.

Determination of total count of fungi:Total counts of fungi were determined on petato dextrose agar (PDA) medium (Christensen, 1957).

pH Values:The value of pH was measured according to (Ling, 1963). pH values of samples were estimated by means of anelectric pH-meter (Wissenschafthich-technic werkstatten D 8120 Weilheimp H40).

Statistical analyses: The data were statistically analyzed  using  GLM  produces   of SAS (1992). Duncan's test was applied in experiment when ever to test differences (Duncan, 1955). The following model was used:

Yij = µ + Ti + eij

Where:Yij = An observed trait, µ= Overallmean, Ti= Effect of  treatment,        eij  =Random error.

RESULTS AND DISCUSSION

Body weight:

            Data present in Table 2 indicated that, body weight of rabbit’s supplemented with 2 and 3 ml/L. NMJ in drinking filter water recorded the highest values than those supplemented with 1 ml/L. NMJ or control diets through the 4 weeks of the experiment. Ibrahiem, et al.,(2004) showed that The live body weight gain was increased by the addition of fresh onion and / or garlic at different inclusion rate.

Table 2. Initial and final body weight of growing rabbits as affected by supplemented with different levels of natural mixture juic (NMJ).          

- Means bearing different letter superscripts (a, b, c and d) within the same row are significantly  (P ≤ 0.05) different.

- NMJ = Natural mixture juice garlic,onion and  lemon.

           The obtained results and recorded observation clearly focused in group received ration supplemented with 3% garlic and 1% onion plus 3% garlic in feed boiler Muscovy duck. Similar finding were reported by El Afify (1997) who found that live body weight and live body weight gain were significantly improved in broilers chickens when fed diet containing onion or garlic extract or garlic residue. El Nawawy (1991) who reported that body weight gain of boiler chickens were improved when adding 1% fresh onion or garlic to their rations, also El Nahla (1983) who stated that adding 2% dry onion or garlic into broilers improved their live body weight gain. Garlic registered a statistically significant increase in body weight of chiken in comparison to the control group (Kumar, et al., 2010).Ahmed et al. (2009) showed that, natural feed additive of 2.5%, significant (P<0.05) increasing body weight compared with the control group in colves. On the contrary, Yalcin et al. (2007) reported that the body weight of chiken was unaffected by the dietaryGarlic powder.

Feed intake:

            Results of the feeding trial  Table 3 indicated that there were an increase in daily feed intake with increasing the level of  NMJ  from 1^st week to 3^rd week of feeding, but the differences were significant. In the 4th week, the rabbits recieved 3 ml NMJ/L  in drinking water recorded the highest value of feed intake followed by 2 ml/L and 1 ml/L, while rabbits of control recorded the lowest value. These results  were in close agreement with finding of Onu and Aja (2011), who showed that the Garlic and Ginger supplementation produced significant (P<0.05) effects on feed intake of the rabbits. Also,  agreement with Alm EI-Dien ( 1999 ) found that Isa-Brown and Fayoumi hens fed garlic in diets their consumed significantly more feed than control. Feed consumption was significantly more when hens fed garlic for 6 than for 3 days/week in diet. This effect was disappeared directly after feeding the basal diet.. The present Our results also, agree with that obtained  by EL Afify( 1997), El Nahla(1983). Also, El Nawawy ( 1991) reported that feed consumption of broiler chickens fed garlic diet was higher by about 6% than the control.Also Ahmed et. al. (2001)  mintioned that juce of lemon, onion and garlic supplemented with different levels (0, 2.5, 5.0 and 7.5%) did not significantly affect the feed consumption of growing buffaloe calves along the feed trial compared with the control, but animal which fed on diet of 2.5% natural juice tended to be good with the highest value.Also, Zaki et al. (2001) found similar results. On the other hand, The present Our results disagree with Ibrahiem, et al.(2004) observe a decrease in feed consumption in all supplemented groups and especially in groups received diet supplemented with 3% garlic or 1% onion + 3% garlic (5.2% - 4.6%) respectively. Horton et al. (1991a) found no obvious effect in feed consumption, when broilers fed powder garlic at the level of 0.1, 1.0 or 10.0 g/kg diet during 1-35 days of age and  El-Habbak et al.(1989) who reported a significant decrease in feed consumption at the 3rd week of age when Japanese quail were fed garlic cloves in diet.Yalcin et al. (2007) reported that feed consumption of chiken was unaffected by the dietarygarlic powder. They explained that the strong odor of garlic does not act as a deterrent of feeding. In anther hand, Ali Nobakht (2013), Mourao et al. (2008) and Rizal et al. (2010) showed that the starter period adding 4.5% Dried Lemon Pulp and in grower and whole periods inclusion 1.5 to 4.5% Dried Lemon Pulp increased significantly the amounts of daily feed intake (P<0.05) and agreement with report of  Chaudry et al. (2004).

Daily weight gain:

Data in Table 3 cleared that daily weight gain of rabbits recieved 3 ml/L,  2 ml/L or 1 ml/L grew faster than control, respectively. These  incorporation of such vegetable and fruitage juice mixtures had no adverse effect on palatability and voluntary feed intake in rabbits. So, introducing different levels of NMJ significantly increased daily weight gain. In this recpect, Ahmed et al. (2009)

Table 3. Daily feed intake and weight gain of growing rabbits as affected by supplemented with different levels of natural mixture juic (NMJ).     

-  a, b, and c = Means within the same row beaing different letter superscripts are significantly  (P ≤ 0.05) different.

pointed that, natural feed additive of 2.5%, significantly (P<0.05) increased daily weight gain compared with the control group,These results might bedue to the effective to improve immunity and decreased ability incidence. Onu and Aja (2011) showed that garlic and ginger supplementation produced significant (P<0.05) effects on weight gain of the rabbits.Also, Ibrahiem, et al. (2004) mentioned that the daily weight gainof broiler Muscovy duckwas increased by the addition of fresh onion and / or garlic at different rate in group received ration supplemented with 3% garlic and 1% onion plus 3% garlic. Similar finding were reported by El Afify (1997) who found that live body weight and live body weight gain weresignificantly improved in broilers chickens when fed diet containing onion or garlic extract or garlic residue. El Naway (1991) who reported that body weight gain of boiler chickens were improved when adding 1% fresh onion or garlic to their rations, also, El Nahla (1983)who stated that adding 2% dry onion or garlic into broilers improved their daily weight gain.Ali Nobakht (2013),showed insignificantly increased the amount of dailyweight gain in starter period  adding dried lemon pulp whole experimental periods (P<0.05). Whereas they had no significant differenceamong groups about daily weight gain in grower period the result agreementwith Rizal et al. (2010), whereas not in agree with Mourao et al. (2008).

Feed conversion ratio:

There were significant (P<0.05) differences among treatments in feed conversion ratio through the first 3 weeks Table 4. Rabbits supplemented with 1, 2 and 3 ml/L. NMJ recorded better FC than the control. The difference was not significant  among the experimental tested groups at 4 weeks of age . These results were similar to those obtained by Ahmed et al.(2009) who revealed that, growing buffaloe calves supplemented with 2.5% natural juice  showed the best feed conversion, but the differences among all groups were not significant. Ibrahiem,et.al.(2004)Showing that the supplementation of different levels of fresh onion and/or garlic to diets of broiler Muscovy duck (12^thweek) areimproved that feed conversion ratio. Group fed supplemented diet with 3% garlic or 1% onion + 3% garlic showed the better feed conversion ratio (2.99 or 3.05) at 10^Thweek and more economically while it was (3.67 or 3.74) at 12th week respectively. Bidura (1999) found that there is increase significantly (P≤0.05) in feed conversion ratio with increasing levels of garlic leaf meal in growing duck diet aged 0 – 8 weeks. The results might be due to the improve immunity and decrease debility incidence, which agree with the finding of Aboul-Fotouh et al. (1999) and (2000), Safaa (1999) who reported that, nutrition plays important role in diminshing growth  rate. Abdo  (1998) observed better feed conversion by using garlic in broiler diet, feed conversion was 1.61, 1.79 and 1.81 for 6,  3 and 0% garlic groups respectively.Onu and Aja (2011)  showed that garlic and ginger supplementation produced significant (P < 0.05) effects on  feed conversion ratio of the rabbits. These improvements may be attributed to the properties of this material that act not only as antibacterial, antiprotozoal and antifungal but also as antioxidant (Bradley, 1992 and Anwar and Meki, 2003). These results are in agreement with Dafwang et al. (1985) who demonstrated improvement in feed conversion ratio of chicks due to decrease in small intestine thickness resulted from feeding dietary antibiotics.

Table 4. Feed  conversion  ratio and performance index  of  growing  rabbits  as ffected by supplemented with different levels of natural mixture juic (NMJ).

-  a,b,c,d = Means within the same row beaing different letter superscripts are significantly  (P ≤ 0.05) different.

.- Feed conversion= Feed intake/ weight gain    - Performance  index%= (Body weight (kg)/ Feed conversion ratio)

On the contrary, Yalcin et al. (2007) reported that feed consumption and feed efficiency of chiken were unaffected by the dietarygarlic powder. While El Afify (1997) who found slight insignificant improvement in feed conversion ratios associated with adding garlic or its fractions into chick’s diets, the value of feed conversion ratio for entire experiments periods (0 – 5 weeks) were 1.83, 1.88, 1.86, 1.89 and 1.91, respectively for minced garlic, garlic juice, garlic residue, garlic oil and control group. Improvement in feed conversion rate attributed to the reduction of small intestine thickness since the nutrient absorption is more efficient through thinner intestinal wall that reflected on the improvement of absorption which translated to improvement in feed conversion.

Performance index (PI%):

          The performance index results of rabbits  supplmented with 2 or 3 ml natural jucie in drinking water  showed significantly (P<0.05) higher values than those supplemented with 1ml and control Table 4. Performance index values were increased by (28.38, 62.42 and 35.15%) for 1, 2  and 3 ml /L. drinking filter water natural juice, respectively than those of the control. Ibrahiem  et al,(2004) reported thatthe better performanceswere obtained when broiler Muscovy duck were fed a diet supplemented with 3% garlic(104.9%) or 1% onion +3% garlic(101.6 %) as both of them consumed less feed and achieved high final live body weight than other groups.

Carcass characteristic:

          Table 5 showed the carcass characteristics of slaughtered rabbits.The differences in carcass weight traits as a results to the effect of adding natural juics of vegitables (garlic and onion) and fruitage (limon) mixture to rabbit diets were significant (P<0.05). Under the condition of this experiment, it appears that, the highest values for dressing % was recorded with rabbits in groups of 3 and 2 ml/L./NMJ in drinkin filter water compared to those had  1 ml/L NMJ. Or the control.

 The results of offals (spleen, two kidneys, liver, heart and lung weight) of rabbits supplemented with 3 and 2 ml/L. in  drinking filter water NMPJ have the same trend of the previous results. The data were in agreement with those reported by Raeesi et al. (2010) who reported that, carcass yield was higher in birds fed 1% garlic powder than those received 0.5 and 3 %. On the contrary EL Afify, (1997) showed that liver weight of broiler were decreased by feeding onion and/or garlic supplemented rations with 11%, 6%, 8% and 11% for groups of chicks fed fresh garlic, fresh onion, dry garlic and dry onion respectively. kamal, (2011) mentioned that, supplementation of Garlic powder had non significant effect on broilers  carcass weight or dressing percentages. On other hand, these results disagreements with those obtained.  This may be due to inhibition of fatty acids synthesis and other lipid components in liver and reduce the level of fat accumulation in liver, which leads to a decrease in liver weight.

Mortality rates:                   

Table 6 represents the results of supplemental NMJ treatments on mortality percentage of growing rabbits. No significant differences among the different treatments were found in the mortality percentage of growing rabbits

from 1 to 4^Th weeks of age. However, adding 3 and 2 ml/L NMJ in drinking water improved viability of growing rabbits. On the other hand, a relatively low mortility rate (10.90 and 12.73%) was observed in rabbits with 3 and 2 ml/L.NMJ  in drinking filter water, while rabbits in  control group recorded the highest mortality rates (21.818%) during the whole experimental period. Similarly in

Table 5. Dressing percentage and internal organs weight of growing rabbits as affected by supplemented with different levels of natural mixture juic (NMJ).     

  a,b,c,d = Means within the same row beaing different letter superscripts are significantly  (P ≤ 0.05) different.

Table 6: Post weaning mortality rate of growing  rabbits as affected by filter water  supplemented with different levels of NMJ * (Means ± SE).

  a,b,c,d = Means within the same row bearing different letter superscripts are significantly  (P ≤ 0.05 or 0.01) different

broilers, it was reported that garlic as a natural feed additive decreased mortality rate (Tollba and Hassan, 2003 and Kamal, 2011).

Microbiological quality of supplemented drinking water:

Table 7 showed the mean of aerobicplate count, total fecal coliform,  total coliform and total Enterobacteria conunt  indrinking filter water with or without natural additive. The mean aerobicplate count of water was (105x10^-3) before  addition, while it was ( 24x10^-5, 6X10^-5and 16x10^-4) with adding 1, 2 and 3%, respectively of the  natural additive. Total aerobic counts were not higher than the recommended safety. The mean total aerobic count for water analyzed is 7.3 x 10-3(cfu/g). For instance, all samples having total aerobic  counts (T.A.B.C.) less than the recommended safety limit of 10^-4 cfu / g proposed by the International Dietetics of Association of European Community (IDAEC) and the Egyptian Organization for Standardization. (2004). Ahmed et al. (2009) pointed that, the mean aerobicplate count of water was (90 x10-2, 320 x10-2, 36 x 10-2 and 77 x  10-2), respectively before addition of the natural additive. These findings are consistent with the quality of wate rexamined in drinking filter water,  where  the   mean  total  aerobic  count obtained in various concentration levels were less than that without additive. The highest concentration level was the least than other concentration levels. The growth of the bacterial strains in different supplemented water was less or not detected after the inhibition effect of the juice at various concentrations.  The three levels of juice inhibited  the bacterial growth and total coliform; total feacal coliform counts; total enterobacteriacea counts and E.coli 01507.  Natural additive contains garlic, onion and lemon juice, garlic contains 0.3-0.5allicin and antimicro component (Shelef, 1993). According,to Kumar and Berwal (1999) and  Zaika and Kissinger, 1983)the gram-positive are generally more sensitive to allicin than gram-negative bacteria. Acetic acid bacteria are the most resistant among the gram positive bacteria. Abdou et al. (1972) concluded that 5-10% fresh garlic was sufficient to inhibit the growth of E.coli. The same trend was observed inonion. Total yeast count were increased with increasing the level of supplementation of natural juice but the total fungi counts were decreased. Also, natural juice did not show any other antimicrobial effect on

Table 7: The effect of different levels feed additive NMJ on determined growth microbial strain of drinking filter water on growing rabbits.

 T.A.B.C.=Total aerobicbacterialcounts, T.C.C.= Total Coliform counts; F.C. C=  feacal coliform counts; T.Y.C. =  Total yeast count;   T.F.C. = Total fungi counts; Total EnterobacteriaceaCounts = T.E.B.C

Salmonella and E. coli. The present results agree with Gherbawy, (1989). Shiva Kumar, et al. (2010) observed the supplemented of garlic active based growth promoter to broiler diets was able to reduce the Salmonella and E. coli. counts in the intestine when in compersion to the negative control and improve growth performance of  broiler.

Conclusively, the results obtained can be recommended that supplements of natural additive mixture of juice, garlic, onion and lemon (NMJ) can be used up to 3ML / L. in filtered drinking water, which gave good results without any adverse effects on growth performance and the best decrease mortality rate of rabbits. Can be used as additives, natural lemon juice, onion, garlic successfully and be safe in drinking water filtered to further improve the performance of rabbits. This study demonstrates that the must be many of the additional studies to further studies are required to more investigate the effect of the natural additive.

REFERENCES:

Abdo, Z. M. A. (1998). The effect of using some natural growth promoter and fats on broiler performance and immunity. Ph. D. Thesis, Faculty Of Agriculture, Animal production department, Cairo University, Giza, Egypt.

Abdou, I. A., A. A. Zeid, M. R. El-Sherbeeny and Z. H. Abou-El-Gheat (1972). Antimicrobialactivitiesof Allium sativum, Allium cepa, Raphanus   sativus, Capsicum frutescens, Eruca sativa, Allnom kurral on  bacteria.  Qualitus   plantarum   et.  Materiae vegetables,22: 29-35.

Aboul-Fotouh, G. E., S. M.Allam,E. I. Shehata  and Abd El-Azeem, S. N. (2000).Effectofsomemedicinal plants as feed additives on milk production and composition  of  lactating  buffaloes.  Egyptian  J. Nutrition and Feeds, 3(1): 31-41.

Aboul-Fotouh, G. E., S. M. Allam, E. I. Shehata  and Abd El-Azeem, S. N. (1999). Effect of somemedicinal plants as feed additives on performance of growing sheep. Egyptian J. Nutrition and Feeds, 2(2):79-87.

Ahmed, A. A.; I. Neamat Bassuony; Set El-Habiab S. Awad; A. M. Aiad and S. A. Mohamed (2009). Adding Natural Juice of Vegetables and Fruitage to Ruminant Diets(B) Nutrients Utilization, Microbial Safety and Immunity, Effect of Diets Supplemented with Lemon, Onion and Garlic Juice Fed to Growing Buffalo Calves. World Journal of Agricultural Sciences, 5 (4):456-465.

Ali  Nobakht(2013). Evaluation  the effects of different levels of  dried lemon (Citrus aurantifulia) pulp on performance of broilers and laying hens. International Research Journal of Applied and Basic Sciences © 2013 Available online at www.irjabs.comISSN 2251-838X / Vol, 4 (4): 882-888 Science Explorer Publications.

Ali, M. and Thomson M. (1995). Consumption of a garlic clove a day could be beneficial in preventing thrombosis. Prostaglandins Leukot Essent Fatty Acids; 53:211–2.

Ali M., Al-Qattan K. K., Al-Enezi F., Khanafer R. M. and Mustafa T. (2000). Effect of allicin from garlic powder on serum lipids and blood pressure in rats fed with a high cholesterol diet. Prostaglandins Leukot Essent Fatty Acids; 62:253–9.

Alm E l-Din,  A.  K.  (1999).  Physiological studies on cholesterol in laying hens.  M.Sc. Theses, Faculty of Agriculture Animal Reproduction Department, Cairo Univ. Giza , Egypt.

Anwar, M. M. and  Meki, A. R. (2003). Oxidative stress in streptozotocin- induced diabetic rats: effects of garlic oil and melatonin. Comp. Biochem Physiol A Mol Integr Physiol., 135:539–547.

A.P.H.A. (American Public Health Association), AWWA (American Water Works Association, WEF. (2005): Standard Methods for the Examination of Water and Wastewater. 21th ed. Washington, DC.

Banerjee S. K, Maulik S. K. (2002). Effect of garlic on cardiovascular disorders: a review. Nutr. Journal; 1:4.

Bolton, F. J., L.Croziev and J. K. Wiliamson,  (1995). Optimization of methods for isolation of Escherichia coli 0157:H7 from beef burgers PHL Smicrobiol. Digest, 12:67-750.

Bradley, P.R. (1992). British  herbal  medicine association.  British herbal compendium, Rolnic zych., 1( 434): 395-399.

Bidura, I. G. N. G. (1999). The effect of garlic ( Allium sativum) leaf meal in diets on performance of growing indoneasian ducks. Majalah Ilmiah Peternakan(Indonesia),.2(6):48-53.

Christensen, G.M.,(1957). Deterioration  of stored grouius fungi.  Botan. Rev.,  23: 108-134.

Chaudry M. A.; Badshan A.; Bibi N., Zeb A.; Ahmed T.; Ali S.and  Termeulen U. (2004). Citrus waste in poultry rations. Arch Geflu¨gelk., 68: 206-210.

Difco,  (2000). Difco manual of  dehydrated culture mediaandreagentsfor microbiologicalandclinical laboratoriesproducts. Ninth Edition Difco Laboratories, Detriot Mi.: Change, USA.

Duncan,D.B., (1955).MultipleRangeandMultiple- Test. Biometrics,11:142

El-Habbak, M. M.; Saleh, K.; Arbid, M. S.; Hegazi ,A. G. and Sofy ,H. (1989). Influence of garlic (Allium Sativum L.) on some biological and biochemical changes in Japanese quail with special reference to its hypocholesterolemic activity. Archive-fur Gefluelkunde. (53)2: 73.

El-Nahla, A. M. M. (1983). Effect of some feed additives on blood constituents and growth rate in chickens.  M. Sc. Thesis, Faculty of Veterinary Medicin, Cairo University Egypt.

El-Nawawy, G. H. (1991). Some  of  non conventional ingredients  in broiler  ration.  M. Sc. Thesis, Ainm. Prod. Dep. Fac,. Agric., Ain-Shams Univ.

El-Afify,  S. F. (1997). Nutritional studies on onion and garlic supplement to poultry feed Ph. D. Thesis, Anim., Prod. Dep. Faculty Agriculture, Ainshams Uuiv.

European Directorate for the Quality of Medicines and Healthcare (2008). The European Pharmacopoeia, 6th ed., Supp. 1, 4-1-2008, online. European Directorate for the Quality of Medicines and Healthcare, Council of Europe, 226 Avenue de Colmar BP907-, F-67029 Strasbourg Cedex 1, France.

Gherbawy,Y. A. M. H.,(1989).Studies on the mycoflora of  pens and feedstuffs of cattle and  chickens in Upper Egypt. M.Sc. Thesis, Bot. Dept., Fac. Sci. Qena. AssuitUniv., Egypt. Egypt. J. Microbial. Sci., 75(1):52-55.

Goorgola, D. L. and M. Boothroyd, (1965). A system for detection  salmonella in meat and meat product.  J. Appl. Bacteriol., 28:206.

Guarrera, P. M. (1999). Traditional antihelmintic, antiparasitic, and repellent usesof plants in centeral Italy. J. Ethnopharmacol.,  68,183-192.

Horton, G.M.I.; Fennell, J. and Prasad, B.M. (1991a). Effects of dietary garlic (Allium Sativum) on performance, carcass composition and blood chemistry changes in broiler chicken. Can.J.Animal Science, 71: 939 – 942.

Ibrahiem, A. Ibrahiem; Talib A. Elam;  Fathi F. Mohamed;, Sabry A. Awadalla; Yousif, I. Yousif (2004). Effect of onion and / or garlic as feed additives on growth performance and immunity in broiler Muscovy ducks. 1^st  Ann. Confr. , FVM., Moshtohor, Sept, 2004, 236-247.

ISO- 6887 (2001). Microbiology  of food and animal feed ingstuffs preparation of test samples, initial suspension and decimal dilutionsfor microbiological examination.

Kamal,  J.(2011). Performance  and lipid  profile  of  broilers fed two  medicinal  plants. Master in Animal Production,  Faculty  Of  Graduate Studies, An-Najah National University, Nablus, Palestine.

Kumar, M. and Berwal, J.  S. (1999).Sensivity of  food  pathogens to garlic (Allium sativum). 3^rd  of   Appl. Microbiol., 84: 213-215.

Kumar S, Sharadmma KC, Radhakrishna PM (2010). Effect  of  a garlic active based growth promoter on growth performance and specific pathogenic intestinal microbial  counts  of  broiler chicks. Int. Journal Poult Sci., 9: 244-246.

Khan, N. A. and A. D. Mcaskey, (1973). Incidence of salmonella in commercially prepareds  and wiches for the vending trades. J. Milk Food Technol., 39:315.

Line-Eric, J. J. Stan Bailey, C. A. Nelson, S. J. Norman and T. Thomas, (1998).  Effect of yeast supplemented  feed   on   Salmonella    and   company   lobacter population in broilers. Poultry Sci., 77:405.

Ling, E. R., (1963). In a Text Book of Dairy Chemistry. Vol.2 Practical 3rdEd. Chapman and Hall.Ltd London, UK.

MacConkey, A. (1905). Lactose-fermenting bacteria in feces. J. Hyg.. 5:333-378.

Mahesar H.; Bhutto M. A.; Khand A. A., Narejo N. T.( 2010). Garlic used as an alternative medicine to control diabetic mellitus in alloxan-induced male rabbits. Pak. J. Physiol; 6 (1).

Mahmoud.  K.; Gharaibeh , S. and Qatramiz K. (2010). Effect of garlic (Allium sativum) supplementation on egg quality and yolk cholesterol in layer hens. Asian-Aust. J. Anim. Sci.,   23 (11): 1503.

Mourao J. L., Pinheiro V. M., Prates A.M., Bessa R. J. B., Ferreira L. M. A. (2008). Effect of dietary dehydrated pasture and citrus pulp on the performance and meat quality of broiler chickens. Poult Sci., 87: 733-743.

Muhammad J., Durrani F., Hafeez A., Khan R.U. and Ahmad I. (2009). Effect of aqueous extract of plant mixture on carcass quality of broiler chicks Journal of Agriculture and Biological Science, 9: 56-59.

Mulumebet Worku, Franco  R., Baldwin  K. (2009).  Arch. Biol. Belgrade, 61(1), 135-140.

Neamat I. Basuony (2004).  Microbial  contamination of fresh meat and poultry during home handling. Thesis D. Ph., Faculty Of Agriculture Alexandria University, Egypt.

North, M. O. (1981). Commercial Chicken Production. Animal 2^nd Edition, AV., Publishing Company I.N.C., Westpost Connecticut, USA.

Olsson, M.E.; Gustavsson, K.E.; and Vågen, I.M. (2010). "Quercetin and isorhamnetin in sweet and red cultivars of onion (Allium cepa L.) at harvest, after field curing, heat treatment, and storage". Journal of Agricultural Food Chemistry, 58 (4): 2323–2330.

Onu P.  N. and Aja P. M. (2011). Growth performance and haematological indices of weaned rabbits fed garlic (Alium sativum) and ginger (Zingiber officinale) supplemented diets.  International Journal of Food, Agriculture and Veterinary Sciences ISSN: 2277-209X (Online) An Online International Journal Available at http://www.cibtech.org/jfav.htm 2011 Vol. 1 (1) October-December, pp.51-59/Onu and Aja Research Article, 51.

Onu, P.N., (2010). Evaluation of two herbal spices as feed additives for finisher broilers. Biotech. In anim. Husb. 26(5-6), p 383-392.Onu P. N and P. M., Aja (2011).Growth performance and haematological indices of weaned rabbits fed garlic (Allium sativum) and ginger (Zingiber officinal ) supplementted diets. Journal Food,Agric.and Veter.Science, 1(1),pp.51-59.

Oxpoid Manual (2000). Oxpoid Manual of culture Media Ingredients and other Laboratory Services. 4^th  Oxoid Ltd).

Prior, R.L., (2003).Fruit and vegetables in the preventation of cellular Oxidative damage.  Am. J. Clin.Nutr.,78: 570S-578S.

Pervez A (1992). Response of broiler chicks to different feed additives. M.Sc Thesis NWFP Agricultural University, Peshawar, Pakistan.

Raeesi, M., Hoeini-Aliabad, S.A., Roofchaee, A., Zare Shahneh, A. and Pirali, S. (2010). Effect of periodically use of garlic (Allium sativum) power on performance and carcass characteristics in droiler chickens. World Acad- emy of Science, Engineering and Technology, 68: 1213- 1219.

Rees LP, Minney SF, Plummer NT, Slater JH, Skyrme DA.A. (2005).  quantitative assessment of the antimicrobial activity of garlic (Allium sativum). World Journal Microbiol Biotechnol; 9:303–7.

Rizal Y., Mahata M. E., Andriani M., Wu G. (2010). Utilization of juice wastes as replacement in broiler diet. Int. Journal Poult Sci., 9: 886-889.

Safaa Nadi, (1999).The use o f  some medicinal plants in ruminant nutrition. Ph.D.Thesis, Faculty of Agriculture, Fayoum, Egypt

SAS, (1992). User's guide: Statistics.,  SAS Inst., Inc.,Cary,Nc.

Shelef, I. A., (1993). Antimicrobial effect of spices. J.of Food Safety, 6:29-44.

Shiva Kumer, K. ; C. Sharadamma and P. M. Radhakrishna (2010). Effect of a garlic active based growth promoter on growth  performance and specific pathogenic intestinal microbial  counts of  broiler  chickes. International Journal Of Poultry Science, 9(3):244-246.

Slimestad, R.; Fossen, T.; Vågen, I. M. (2007)."Onions: a source of unique dietary flavonoids". Journal of Agriculture and Food Chemistry, 55 (25): 10067–80. 

Thomson  M., Al-Amin Z. M., Al-Qattan K. K., Shaban L. H., Ali M. (2007). Anti-diabetic and hypolipidaemic properties of garlic (Allium sativum) in streptozotocin-induced diabetic rats. Int. Journal  Diabetes Metabolism, 15:108–15.

Toulah, F. H. and M. M. Al-rawi (2007). Efficacy of garlic extract on hepatic coccidiosis in infected rabbits (Oryctolagus cuniculus): Histological and biochemical studies. J. Egypt. Soc. Parasitol., 37(3),957-968.

Tollba, A.A.H. and M.S.H. Hassan, (2003). Using some natural additives to improve physiological and productive performance of broiler chicks under high temperature  conditions. Black cumin  (Niglla sativa) or Garlic (Allium sativum). Poult. Sci. Journal, 23; 327-340.

Yasmin Khan, Rafeeq Alam Khan*, Syeda Afroz and Afshan Siddiq(2010) .Evaluation of hypolipidemic effect of citrus lemon. Journal of Basic and Applied Sciences Vol. 6, No. 1, 39-43.

Williamson, Gary; Plumb, Geoff W.; Uda, Yasushi; Price, Keith R.; Rhodes, Michael J. C. (1997). "Dietary quercetin glycosides: antioxidant activity and induction of the anticarcinogenic phase II marker enzyme quinone reductase in Hepalclc7 cells". Carcinogenesis,17 (11): 2385–2387. 

Zadik, P. M.; P. A. Chapman  and C. A. Siddons, (1994). Immunomagnetic  separation as  a  sensitive  method for isolating Escherichiacoli0157:H7from food samples. Epidemiol. Infection, 113:31-39.

Zaiaka,L.A.andJ.C.Kissinger, (1983).Inhibitoryand stimulatory  effects  of oregano  on  Lactobacillus  planetarium and Pediococcus cerevisiae. Journal  of  Food Sci., 46:1205-1210.

Zaki,   A. A.,   M. R.  Mostafa,   R. T.  Fouad  and Z. M. Marei (2000). Teosint (Zeamexicanal) forage      productivity  quality  and  its  feeding  effect  on performance  of buffalo calves) Proc. Conf. Anim.  Growth Prod. In 21thCenturysakha, 18-20 Apri., pp:1737-244.

Yalcin, S., Ĺ. Onbaşilar, A. Şehu, and S. Yalcin. (2007). The effects of dietary garlic powder on the performance, egg traits and blood serum cholesterol of laying quails. Asian-australas. J. Anim. Sci., 20:944–947.

تاثیر اضافة مخلوط العصیر الطبیعى للبصل و الثوم و اللیمون فى ماء الشرب المفلتر على أداء الأرانب النامیة

د.عزة محمد محمد بدر¹  و د.نعمات بسیونى ¹ و د.فضیلة محمد عیسى² و د.مجدى فؤادسلیمان حنا²

1- المرکز الأقلیمى للأغذیة و الأعلاف بمرکز البحوث الزراعیة – 9 شارع الجامعه - جیزة – مصر

2-  معهد بحوث الانتاج الحیوانى – الدقى – جیزة -  مصر

أجریت هذة الدراسة على 220 أرنب نامى محلى خلیط وکان وزن الجسم الحى  فى بدایه التجربه  751.8+ 22.4 جرام و عند عمر 33 یوم . قسمت الأرانب الى  أربع مجموعات تجریبیة عشوائیا ( کل  مجموعه 55 أرنب). غذیت المجموعات التجریبیة على مخلوط علف مرکز (16.72% بروتین خام و 13.07% الیاف خام ). المجموعه الاولى (الکنترولD¹) غذیت بدون اضافة MNJ        (مخلوط  العصیر الطبیعى المکون من الثوم والبصل و اللیمون)،  بینما أضیف للمجموعات الثانیة( D²)  و الثالثة (D³) و الرابعة ( D⁴)1& 2& 3 مل/ لتر ماء شرب مفلتر على التوالى. استمرت تجربه التغذیه 4 أسابیع ، وکان الهدف من الدراسة هو تقییم اضافة مخلوط العصیر الطبیعى الى ماء الشرب المفلتر الیومى على الزیادة الوزنیة و کفاءة التحویل الغذائى ووزن الذبائح والتصافى ونوعیة المیکروبات فى ماء الشرب.

أشارت النتائج الى أن العلف المأکول الیومى للأرانب قد زاد بزیادة وجود العصیر الطبیعى فى ماء الشرب المفلتر أکثر من مجموعة الکنترول. الأرانب التى أضیف لها مخلوط  العصیر الطبیعى الى ماء الشرب مفلتر بواقع 3& 2& 1 مل/لتر قد سجلت نموا أسرع   بالمقارنه بالکنترول على و سجل وزن الجسم النهائى للأرانب التى أضیف الیها 2& 3 مل / لتر ماء الشرب  المفلتر نفس النتائج السابقة.

کانت هناک اختلافات معنویه(5%) بین المعاملات التجریبیة فى معدل التحویل الغذائى خلال الاربع اسابیع الاولى من التجربة سجل وزن الذبائح و%التصافى للارانب التى اخذت عصیر طبیعى فى ماء الشرب اعلى قیمه معنویة(5%) عن مجموعة الکنترول. بالنسبة الى نوعیه المیکروبات فأن زیادة مستوى مخلوط العصیر الطبیعى فى ماء الشرب المفلتر قد قلل من اعاد البکتیریا الهوائیة والکولیفورم الکلى والانتیروبکتیریا والخمائر والفطریات کما لا توجد بکتیریا الاشیرشیا کولاى فى ماء الشرب المفلتر.

التوصیة : النتائج المتحصل علیها یمکن التوصیة بأن المکملات الغذائیة من خلیط من عصیرالمضافة الطبیعیة والثوم والبصل واللیمون (NMJ) یمکن استخدامه حتى 3 مللى / لتر من ماءالشرب المفلتر، والتی أعطت نتائج جیدة من دون أی آثارسلبیةعلى اداء النمووأفضل انخفاض فى نسبه النفوق الأرانب. یمکن استخدام إضافات،عصیراللیمون الطبیعی،والبصل، والثوم بنجاح وتکون آمنة فی میاه الشرب المفلتر لزیادة تحسین أداء الأرانب. وتوضح هذه الدراسة أن متوسط الزیادة الیومیة کان أکثر مما کان متوقعا ویجب أن یکون العدید من الدراسات الإضافیة لمزید من دراسة تأثیرالمادة المضافة الطبیعیة.