Effect of some Transition Complexes on Bacteria and Fungi Causes Gastroenteritis in Humans

This study concerned with the synthesis and characterization of some complexes of the ligand. All the complexes are non-electrolytes place , instead crystalline solid compounds insoluble in water , but soluble in organic solvents the negatively charge bidentate ligand coordinated with metal ions through the oxygen atom in the hydroxyl group and the oxygen atom of the aldehyde group. The effect of these complexes on inhibiting the growth of bacteria and fungi isolated from diarrheal patients with intestinal inflammation has been shown to be highly inhibitory and has been the best inhibitory effect of the complex Zn(OC10H6CHO)2 on bacteria Shigella dysentariea inhibition diameter 32 milmeter and on fungus candida glabrata in hibition diameter 27 milimeter and there were no toxic effects of these complex on the Laboratory mice are the dose of 2.5 – 15 gm / kg of body weight Keyword: Keyword: Transition, Complexes, Bacteria, Fungi, Gastroenteritis.


Gastroenteritis (GIT )
Is a condition called satisfactory inflammation of GLT , which includes both of the stomach and small intestine , which lead to the total of symptoms such as diarrhea , vomiting , abdominal pain and spasm called gastro esophageal 1 , is causing the most common cases gastroenteritis in children Rota virus and adults Nor virus 2,3 .also may occur because many of bacteria Escherichia coli , Staphylococcus aureus, Salmonella spp, Bacillus spp.,VibrioCholera and other types of bacteria it may happen due to the consumption of foods stomach improperly or because of drinking contaminated water 4 .Gastrointestinal tract is more common among infants and children.They cause of death under 5 years age in the developing world 5 .The intestinal bacteria transmitted by eating contaminated food especially beef and their products are healthy cows main treasure and high rate was observed in rural as a result of direct contact with cow or the faces containing the bacteria 6 .

Synthesis of Ligand
Ligand 2-hydroxy-1-naphthaldehyde (C 11 H 8 O 2 ) it is one of the aromatic compound derivatives of naphthalene dark brown crystalline and the degree of melting 76-80 o C and boiling point 192 o C/27mm Hg (lit) not soluble in water but soluble in organic solvents molecular weigh 172.183 g/mole.

Ligand preparation
Ligand was prepared by the Gattermann-Koch reaction(also known as the Gattermann salicylaldehyde synthesis) is a chemical reaction in which aromatic compounds are formylated by hydrogen cyanide in the presence of a Friedel-Crafts catalyst (e.g AlCl 3 ).It is named for the German chemist ludwing Gattermann and is similar to the Friedel-Craft reaction.
Combination with zinc cyanide.Although it is also highly toxic.Zn (CN) 2 is asoild, making it safer to work with than gaseous HCN, additionally, because the reaction uses HCI,Zn(CN) 2 also supplies the reaction with ZnCl 2 in situ, where it acts as a lewis acid catalyst.
through the oxygen atom in the hydroxyl group ( -OH ) and the oxygen atom of the carbonyl group shown in Figure (1).
Examples of Zn(CN) 2 being used in this way include the synthesis of 2-hydroxy -1naphthaldehyde and Mesitaldehyde preparation of ligand .The Ligand (L) was prepared as follows 7 .
Ligand be charged negatively ( L -) after the loss of a proton ( H + ) from a hydroxyl group ( OH ) bidentate ligand coordinated with metel ion

Collection of samples
Sixty four clinical specimens collected for children and adult , distributed between 20 stool sample adults of both sexes and 44 sample of stool of children less on 10 years of age suffer from diarrhea for the duration of January 2016 until august 2016 from different hospitals in Baghdad , collect stool by wooden sticks and placed in a sterile tube containing 2 ml of saline solution then cultured on selective isolation specific media such as macconkey media , blood agar media and initially diagnosed by examination of morphometric traits developing colonies then microscopic examination by using gram stain 8,9 .Then differential Biochemical test like Idols test, methyl red, vogas proskauer, citrate utilization, urea hydrolysis, triple sugar iron test and diagnosis by growth on cultural media like Eosin methyl in blue media, Salmonella -Shigella agar, mannitol salt agar then to confirm used EPI 20 E and API 20 staph 10 .and diagnosed the fungi by culturing on the sabouraud dextrose agar, CHROM agar 11 .

Preparation of complexes
The chemical was used ligand 2-hydroxy-1-naphthaladehyde and equipped company ( E. Merck) and metal salts equipped from different companies Fluka, Merck & Co., Ridel-de-Haen age and Al-don Chemical Inc.
Complexes were prepared by taking 0.25 gm ( 1 mmol ) from ligand according molar ratios mentioned earlier and has melting this quantity at the lowest possible amount of ethanol (10-15) ML then add salt solution of the dissolved in ethanol between (0.33-1) mmol,(0.1-0.24)gm move mix continuous manner where it observed the emergence of a deposit has been nominated and washed with distilled water and ethanol and then returned crystallized with ethanol and dry temperature 50 o C that some of the complexes needed preparation to a temperature higher than 60 o C and on the water bath they both complexes [Mn(OC 10 H 6 CHO) 2 ],[Ni(OC 10 H 6 CHO) 2 ] and attended all the complexes in the Neutral surroundings complexes of 2-hydroxy-1-naphaladehyde with some metal salt 12 .

Study the effect of trans elements compounds on growth of bacteria
Diffusion method used in the agar the note the effect of chemical compounds on the growth of bacteria isolated.Cultured Muller Hinton agar in with sterile swab.loaded bacteria containing 1.5 × 10 8 cell / mL , worked wells on the media ager by cork borer and put the prepared concentrations of each compound by 50 micrometer by each well with a DMSO ( Dimethyl sulfoxide ).Control to confirm that the inhibitory has no effect on the growth of bacteria and leave the microletter in the laboratory temperature for 15 min.then incubated a temperature 37 o C in 24 h.and an average of three replicates for each isolated and Identified the complexes effectiveness by measuring the diameter area around each hole and the conference has been three replicate 13 .

Study the effect of trans element on the growth of fungi
Fungi prepared from suspension fungal and cultured on the sabouraud dextrose agar and worked in with holes by cork borer and put trans elements compounds in the well and petridishes were let 15 min.at laboratory temperature and then incubated 24 h at 37 with three replicates of each concentration of the transelement and read results by measuring inhibition zone of bacteria around the wells 14 .

Minimum inhibitory concentration ( MIC ) and minimum bacteriocidal concentration ( MBC ) of trans element complex.
The method of turbidity was used 15 .In addition to the control tube, add 0.8 mL of sterile brain heart infusion broth (media of bacteria) and other sterile sabouraud dextrose broth (Media of fungi ) , to the small test tubes .then add 0.1 mL of the concentration of the selective elements except the control tube and then add 0.1 mL of the bacterial suspension compare with Mac far land tube number 0.5.The tubes were incubated at 37 o C for 18-24 hours.The result were coloured on the basis of the turbidity observation and 100 mew L were taken from the bacterial mixture or fungal mixture and incubated for a period of 24 h and temperature 37 o C and recorded the result on the basis of the presence of growth number of colonies or lack of growth.

Lethal dose of moderation of transelement complexes on the laboratory mice by mouth dosage.
We have used 35 Swiss Webster albino mice for each trans element compound and dosage gradual dose (2.5,5,7.5,10,12.5,15)gram /kg of body weight with total control and repeated the experience the same number of other mice were observed during 96 hours 16 .

RESULT AND DISCUSSIONE
Out of 87 patients, 64 were found suffering with inflammation of stomach and intestine in the hospitals of Iraq.as well as, seven types of bacteria and four species of fungi depending on microscopic, phenotypic, biochemical test and API staph, API 20 E as it is shown in the following table1.

The effect of the transelements complex on the growth of bacteria isolated from cases of diarrhea
The transelements of the positive results in inhibiting the growth of bacteria and these results are different according to the type of transelements and bacteria type as [ The impact of these trans elements complex may return the effect on bacteria according to their ability to solubility in membranes Lipid so the work inhibitory either be outside the cell or the cell surface or intra cellular cell.The effect of the trans element of bacterial cells in term of their effect on the cell membrane or impermeability so this transformation from an optional permeability to random permeability and thus allow the passage of toxic Material 17 or out material and essential elements of the cell and some of trans elements affect the enzymes produced by the cell, which prevents bacterial work or interference to the cell through the bacterial cell membrane and prevent metabolic reactions in the cell, such as respiratory reactions and thus play elements to the inhibition of bacterial cell growth 18

Table. 4: MIC and MBC of transelement affecting the growth of bacterial and fungal isolates
The trans elements complexes are highly effective on fungi belonging to the types of candida as in the table (3) and Fig 6,7 this could go back to the inhibitory potency of these fungi to effect mitochondria or endoplasmic and plasma membrane and these to inhibit fungus 19 .
The trans elements complex were effective in inhibiting and killing different bacterial and fungal isolates .The best inhibitory agent was [Cd (OC 10 H 6 CHO ) 2 ] on bacteria Enterobacter cloacae , Sh .dysenteriesand candida sp. with concentration 5% it is best to kill the same bacteria and fungus with concentration 10% as shown the table(4) and there is no effective toxicity or loss on laboratory mice exposed to different concentrations of trans element and there are no histological or hemorrhagic effects on laboratory mice at dissection after 96 h at different concentrations as shown figure 8.