The Influence of Tilmicosine and Enrofloxacine in Mycoplasmas Control, Hatchary Performance and Quality of Eggs and Day Old Chicks

The aim of this study is to evidence the role of antibiotics Tilmicosine and Enrofloxacine in mycoplasmas (MG) control, their impact in hatchery performance and quality of eggs and day old chicks in breeder broiler chickens. The experiment took place for 63 weeks in a breeder broiler farm in Ioannina’s area in Greece. From this area Albanian farmers import the meager number of day old chicks. The 3 groups, were positive in M.gallisepticum (Eliza test, Check flock Mg –IDEEX and cultural control). The best results were in the Tilmicosine group, not only in M.gallisepticum control, but also in the hatchability which was 85.6 %, 84.15% and 83.2% respectively for the Tilmicosine, Enrofloxacine and control group. The average of eggs and day old chicks weight in different days of laying curve (day 210, 280, 350) were better in the Tilmicosine and Enrofloxacine groups, than the control group. INTRODUCTION The hatchability in the incubator and quality of eggs and day old chicks in breeder broiler chickens, is a very important indicator for farmers of the parental flocks. Many factors play an important role in the percentage of the hatching of eggs like the genetic strain (Abudabos, 2010) quality of eggs (Yoho et al,2008), age of broiler breeders (Tona et al,2004., Dikmen et al,2007), egg shell and incubation condition (Christensen et al, 2005), nutrition (Ayidin et al, 2004., King’ori, 2010), Environmental factors (King’ori,2011), egg storage position (Tiwari et al, 2005), males and females rapports ( Sapp, 2004), eggs size (Abiola, 2008) etc. Different microorganisms, espetially mycoplasmas have an affect on the fertility and the embryonic deaths and as a result on the percentage of hatching of the eggs in the incubator. (Levisohn et al, 1985., Lockaby et al, 1999., King’ori, 2011) The chick’s vitality, besides others, has also to do with the chick’s weight which is interdependent from the egg’s weight. Lots of authors mentioned that the one day broiler chicks weight, can affect its weight in slaughter house at the age of 38-40 days (Leeson et al, 2000). Besides other factors, the egg’s and hatched chick’s weight, can be influenced by several factors with microbe origin such are Mycoplasmas. In order to see concretely the influence of Mycoplasma gallisepticum in the quality of eggs and hatched chicks, we organized the following experiment: MATERIAL AND METHOD In a breeder broiler farm that produces eggs for broiler incubation, there have been created three groups with 3500 female chicks on each. In the two of the three groups we used the antibiotic Tilmicosine and Enrofloxacine in the first 3 days that the chicks just entered in the stable with dose 30ml Tilmicosine 25% in 100L/drinking water (Pulmotil AC, Elanco) and 50ml of Enrofloxacine 10% in 100L/drinking water(Baytril, Bayer). Then, Tilmicosine was used for 5 other times for one day with dose 20mg/kg.b.w with four-week intervals till the beginning of the eggs’ lay. The same scheme but for 2 days was using the Enrofloxacine 10mg/kg body way. Upon the beginning of the eggs’ lay. No more antibiotics were used till the removal of the breeder stock from the stable which lasted for 63 weeks. In the control group no antibiotic was used. Before the antibiotics usage, 15 chicks for every group were made the blood infusion in specified days and the control for the presence of Mycoplasma antibodies was made, through the ELIZA’s test, Flock check MG-IDEEX. There were also made cultural controls in determined days in respective cultures. The cultural tests have taken place in microbiology laboratory of the Faculty of Medicine in Iannina-Greece. The terrains PPLO agar and bujon medium of the company Difco were used , adding the supplement Mycoplasmas: yeast extract, horse serum, penicillin and thallium acetate according to the assigned methodology. From the laboratory, clinical and anatomy-pathological check, we had no other microbe factors to affect the experiment result. But however, the conditions in the three experimental groups were equal. We collected the eggs for each group and put in the incubator seperately under strict control on every detail. Then we scaled with electronic scale by 100 eggs taken occasionally from each group, and we got out the average for one egg, placing them separately for incubation. The chicks hatched by these eggs were scaled with the electronic scale, separately for every group getting out by this way the average for one chick. For this purpose, the eggs have been taken in assigned days that coincided with the peak of the eggs’ production (lay). On the 210th day on which the eggs’ production was 88%, later on the 280th day on which the eggs’ production was in the middle of the production curve over 70%, and at the end of the production period on the 350th day on which the percentage of eggs’ lay was about 50%. RESULTS AND DISCUSION In table no.1 are presented the results of serological analysis.


INTRODUCTION
The hatchability in the incubator and quality of eggs and day old chicks in breeder broiler chickens, is a very important indicator for farmers of the parental flocks. Many factors play an important role in the percentage of the hatching of eggs like the genetic strain (Abudabos, 2010)  The chick's vitality, besides others, has also to do with the chick's weight which is interdependent from the egg's weight.
Lots of authors mentioned that the one day broiler chicks weight, can affect its weight in slaughter house at the age of 38-40 days (Leeson et al, 2000).
Besides other factors, the egg's and hatched chick's weight, can be influenced by several factors with microbe origin such are Mycoplasmas.
In order to see concretely the influence of Mycoplasma gallisepticum in the quality of eggs and hatched chicks, we organized the following experiment:

MATERIAL AND METHOD
In a breeder broiler farm that produces eggs for broiler incubation, there have been created three groups with 3500 female chicks on each. In the two of the three groups we used the antibiotic Tilmicosine and Enrofloxacine in the first 3 days that the chicks just entered in the stable with dose 30ml Tilmicosine 25% in 100L/drinking water (Pulmotil AC, Elanco) and 50ml of Enrofloxacine 10% in 100L/drinking water(Baytril, Bayer). Then, Tilmicosine was used for 5 other times for one day with dose 20mg/kg.b.w with four-week intervals till the beginning of the eggs' lay. The same scheme but for 2 days was using the Enrofloxacine 10mg/kg body way. Upon the beginning of the eggs' lay. No more antibiotics were used till the removal of the breeder stock from the stable which lasted for 63 weeks. In the control group no antibiotic was used. Before the antibiotics usage, 15 chicks for every group were made the blood infusion in specified days and the control for the presence of Mycoplasma antibodies was made, through the ELIZA's test, Flock check MG-IDEEX. There were also made cultural controls in determined days in respective cultures. The cultural tests have taken place in microbiology laboratory of the Faculty of Medicine in Iannina-Greece. The terrains PPLO agar and bujon medium of the company Difco were used , adding the supplement Mycoplasmas: yeast extract, horse serum, penicillin and thallium acetate according to the assigned methodology. From the laboratory, clinical and anatomy-pathological check, we had no other microbe factors to affect the experiment result. But however, the conditions in the three experimental groups were equal.
We collected the eggs for each group and put in the incubator seperately under strict control on every detail.
Then we scaled with electronic scale by 100 eggs taken occasionally from each group, and we got out the average for one egg, placing them separately for incubation.
The chicks hatched by these eggs were scaled with the electronic scale, separately for every group getting out by this way the average for one chick.
For this purpose, the eggs have been taken in assigned days that coincided with the peak of the eggs' production (lay). On the 210 th day on which the eggs' production was 88%, later on the 280 th day on which the eggs' production was in the middle of the production curve over 70%, and at the end of the production period on the 350 th day on which the percentage of eggs' lay was about 50%.

RESULTS AND DISCUSION
In table no.1 are presented the results of serological analysis.

RESEARCH PAPER
In the second day we don't have the presence of the antibodies of Mycoplasma gallisepticum in the blood serum infused to the newly arrived chicks in stable. This demonstrates that grandparents flock is free of Mycoplasma gallisepticum, under the conditions of a rigorous bio-security. The serological analysis of the 30 th day showed the presence of mycoplasma's antibodies in the blood serum in the control group and Enrofloxacine group, such a fact shows the horizontal infection of chicks because of the lack of a non rigorous bio-security.
In the serological analysis after the 120 th day, we have the presence of mycoplasma's antibodies in all groups. The serological analysis was assisted from the cultural tests too for the increase of MG presence.
In  This increase of mycoplasmas in control group, in our opinion has to do with the non-usage of antibiotics for a long time, and consequently the MG has the possibility to continue its multiplying.
Our results are in contradiction to the conclussions of some authors that have applied the antibiotics usage only in pullets impeding the transmission of mycoplasmas through eggs in the newly hatched chicks (Elanco Italy, 2003). Our results showed that under the conditions of a non-perfect bio-security, the antibiotic usage only in pullets till the beginning of the eggs' lay, is not able to control the mycoplasmas' transmission through eggs in the newly hatched chicks.
These results show that the frequent monitoring of breeder stock is necessary , in order to take precaution for the mycoplasmas control, through the increase of bio-security and through the frequent usage of antibiotics in chickens during the laying period (Barbour et al, 1998., Nascimento et al, 2005). In total the percentage of the hatching in the incubator is obviously different. Which is 73 756 , more hatched birds in Tilmicosine's group in comparisson to control group and 34 750 in comparison to Enrofloksanine.
These results are of a big interest for an economy that deals with egg production and the hatching of birds in incubators. So the economical effect of Tilmicosine should be taken under consideration . From the careful observation of tables' no.4 , it results clearly that the eggs produced from chickens on the 210 th day, which coincides with the peak of egg's production, are smaller, so with average weight that is smaller compared to the weightings on the 280 th and 350 th day. Not only the average weight of their eggs, but also that of the chicks hatched from them, is smaller.
This average weight of both eggs and chicks, increases on the 280 th day and reaches the biggest weight on the 350 th day.
A certain progress of egg's weight, and consequently of the chicks' weight that are hatched from them, is physiological (Leeson et al, 2000., Yanacopoulos et al,1987. The egg weight is in the right correlation with chickens' body weight, which increases continuously till the end of the laying period (Leeson et al, 2000).
The weight of chicks hatched from eggs in the peak of their production, so from small eggs, must be taken into consideration from the importers of our country that debate and require from the farmers of the parent stock of Greece and Italy to receive chicks that come from as big eggs as possible and that chicks have the most possible weight.
However, in every phase of weighting we have differences between the three groups concerning the average weight of the eggs and of the chicks hatched from them. In the group where we have used the antibiotic macrolid Tilmicosine, the weight is bigger compared to the Enrofloxacine group and both eggs from groups treated with antibiotics were bigger than control group. Certainly, in absolute values, this difference seems inconsiderable, but by the biological point of view it has noticeably influenced in the hatch of a more vital chick which can pass easier the initial stresses, the receipt and assimilation of nutrition etc. Furthermore, a vital chick is very resistant towards the different infections, including also the respiratory infections (Dufour-Zavala,1999) such are mycoplasmas which are frequent in poultry industry.
The differences between groups and ages of weighting, is verified also by the results' statistical processing.
Other authors have studies the correlation between the chicks' weight in hatch and those in slaughter house. As big as the weight of chicks is in the hatch phase, as big is it during the growth and also in slaughter house (Halevy et al, 2004).
The difference in the percentage of the incubated eggs' hatch and of the quality of the chick's weight is dedicated to the antibiotics concentration by lots of authors, especially to pharmacokinetics of macrolide antibiotics such as Tilmicosine in the egg, in egg yolk and in the egg embryo controlling well and decreasing the microbe number of Mycoplasmas (Goetting, 2011).
The decrease of Mycoplasmas' number, in our case MG, decreased their negative impact in eggs and in eggs' embryo, increasing so not only the fertility and hatchability, but also the weight and vitality of the chicks hatched from them (Mohamet