The test was conducted at the Experimental Zootechnical Structure of the DIPROVAL – University of Bologna, on 63 weaned piglets (crosses Large White x Landrace), 21 days old, taken from 12 litters. Sex was registered.

Two dosage of Turval B0399:

(recommended dosage) 1,5 g/kg and (overdose: tolerance test) 15 g/kg of feed were used.

All the subjects were slaughtered at the end of the test.

During the experimentation the following parameters were evaluated:

-          growth in post-weaning period,

-          conversion index,

-          values of intestinal pH (duodenum, ileum, colon and cecum),

-          concentration of total coliforms

-          lactobacilli in the cecum.

-          Kluyveromyces in the colon

 

 

Effects of recommended dosage and over-dosage (tolerance test) of Turval B0399 in the diet of the weaned piglet on:

growth in post-weaning period,

conversion index,

value of intestinal pH,

concentration of total E. Coli in the intestine (cecum)

presence of kluyveomyces in the colon.

The test was conducted at the Experimental Zootechnical Structure of the DIPROVAL – University of Bologna, on 63 weaned piglets (crosses Large White x Landrace), 21 days old, taken from 12 litters. Sex was registered. The trial lasted 28 days on average.

All the subjects were slaughtered at the end of the test.

 

 

Breed:                        Large White x Landrace

Age:                           21 day old

Sex:                            mixed, registered for statistical analysis

 Through auricular numbering from No. 1 to No. 63 of the subjects tested.

All of the subjects were tattooed on the ear with the number.

033RE 109 indicated the stock room from which they came.

 

 

 

·      Number of groups:                                                         3

·      Number of groups treated with TurvalB0399: 2

Turval B0399, 1,5 g/kg of feed (T1)

Turval B0399, 15 g/kg of feed (T2 overdose: tolerance test)

 

·      Number of groups of control (C):                                  1

·      Number of animals in each group:                               21

·      Total number of animals (piglets):                                63

·      12 litters were weaned at 21 days on average and groups were formed as follows.

From each litter, 3 to 11 subjects were collected.

From each litter, triplet of subjects similar for the live weight were created and one subject per triplet was assigned at random to each dietary treatment. After this, for litters where residual subjects were present, these piglets were randomly assigned to one or two of the dietary treatments, in order to equalize the total number of subjects. In any case all the litter were represented in each treatment. The overall distribution is reported in the data annex.

The weight and sex was registered.

 

All individual data was elaborated through analysis of the variance factor, utilizing the procedure GLM of the SAS package (SAS/STAT User’s Guide, Vers.6, 4^th ed., Vol.2, Cary, NC, SAS Institute Inc., 1989.). Diet, sex and litter were considered in the model. The number of subjects chosen was statistically sufficient. 

The 2 degrees of liberty of the “diet” treatment were divided in the orthogonal contrasts “Control vs Turval B0399 diets” and “Between Turval B0399 diets”. In substance, in the first comparison the general effect of Turval B0300 was compared with a normal standard diet. With the second contrast, the differences between the 2 dietary levels of the products were tested. However for better information of the reader, 2 other contrasts are displayed: Control vsT1 and Control vsT2, i.e. both the effect of the proposed dose of the products and of the ten times dose are compared to the standard diet.

For the recovery of Turval B0399 in the cecum, the effect of dietary treatment was pested by the Likelihood Ratio Chi-Square Test.

 

Concentration of Turval B0399 in the feed for testing

 

T1: recommended dosage 1,5 g of additive /kg of feed = (5 x 10 ^6 x CFU /g x 1,5 /1000 = 7,5 x 10 ³  CFU Kluyveromices B0399/g of feed

 

T2: over dosage (10 x T1) 15 g of additive /kg of feed = 75 x 10 ³ CFU Kluyveromices B0399/g of feed

·      C   = Control group: feed without additive

·      T1 = Treated group: feed with recommended dosage:

·      T2 = Treated group: feed with overdose (tolerance test).

 

 

Table 1.3.2 - CFU of Kluyveromyces B0399 in the additive.
(cfr: analysis Chemio n. 25700)

 

 

The analysis of the additive is shown in Table 1.3.1

 

 

·      Experimental facility: DIPROVAL Univ. of Bologna, in Reggio Emilia, Italy 
The subjects were reared in individual cages with slattened floor.
The internal temperature was automatically controlled and set at 28 °C (starting day) to 21 °C (final day).
In the 1^st week infrared light lamps placed over each subject supplied additional heating.

·      State of health of the animals at the beginning of the experiment: good

·      Conditions of feeding and breeding: normal

·      Control of feeding and measures taken to prevent the contamination of the groups of reference (especially through cross contamination of the foods caused by microorganisms):
The diets were prepared from the same feed. The feed was produced in a single phase and in a quantity sufficient for sustaining the entire test for all the animals. To obtain the diets with Turval B0399, the basic diet in the quantity of 10 kg at a time, was supplemented with 15 g and 150 g of the additive, respectively for T1 and T2, thoroughly mixed in a plastic container and labeled with the name of the dietetic treatment (name of the lots).

 

 

·      Beginning of test: 11 April 2002

·      Total Duration : 28 days

·      Final weight of reference: 11 kg

·      At the end of testing the animals were slaughtered.

-          Recording weight at the beginning and at the end of the test.

-          The general health of the subjects was also observed.

-          Determination of the pH in various parts of the intestine: duodenum, ileum, cecum, colon.

-          Collection of content of the colon for the total E.Coli and Lactobacilli count.

 

(cf. Table 2.3.B.2)

 

Seven subjects died during the trial, respectively 2 for control group, 1 for T1 and 4 for T2. All the subjects died suddenly and at the necroscopy showed signs of enteritis and of colibacillosis. In 5 cases an Escherichia coli hemolytic strain was isolated; in one case from kidney, liver and brain a Sterptococcus suis was isolated.

 

In Par 9 Annex the individual data and results of the subjects are shown.

In table 2.3.B.2 the results regarding the growth of the subjects are shown. Firstly the average starting weights of all the subjects are displayed. Secondly the average starting weights of the subjects that were in trial till the end of the experiment are presented. In this case, due to the high number of subjects that died in the overdose dietary treatment and to their individual starting weights, a statistically significant difference was observed between T1 and T2 treatments, in favor of the latter. In any case, the diets did not significantly modify the final live weight and the rate of growth, which remained low, on the average 173 g per day,-- a common phenomenon in the first post-weaning week.

The feed intake was not affected neither the feed conversion was influenced by the diet.

The data on growth performance should be evaluated considering the negative health condition of the overdose group. In this case the live weight gain was not affected in the residual subjects, bur, overall, 4 of 21 subjects died for intestinal pathology.

For the standard does group at the contrary no negative drawback was observed and in this group the mortality was the lowest (1 of 21).

In the similar trial 51 B executed by this Department in 1999, a positive effect on growth performance was observed.

The absence of a evident effect of Turval B0399 in this parameter of the present trial, could be attributed to differences in the base diet, that could have affect also the presence of other bacteria aspects not cultivated or not cultivable.

This could have consequently contributed to maintain a similar results between the different diets.

 

 

 

In the similar trial 51 B executed by this Department in 1999, a positive effect on ph was observed.

The absence of a evident effect of Turval B0399 in this parameter of the present trial, could be attributed to differences in the base diet, that could have affect also the presence of other bacteria aspects not cultivated or not cultivable.

This could have consequently contributed to maintain a similar pH between the different diets.

 

 

 

The number of total coliforms found in the colon was reduced. The content of Lactobacilli was increased by Turval B0399 supplementation.

The ratio coliforms/lactobacilli was reduced by both dosages in a statistically significant manner (P<0.01 for T2 and P<0.05 for T1).

With the standard dosage and overdose of the additive, the recovery of Kluyveromyces B0399 was 36.8% and 47.1 % respectively. In the control diet no subject was found positive for the presence of Kluyveromyces B0399. This difference against supplemented diets was statistically significant (Likelihood Ratio Chi-Square Test: P=0.001).

 

 

 

 

The trial showed that the recommended dose dietary addition of Turval B0399 did not negatively affect the growth performance of weaned piglets, the health and the pH of the different intestinal tracts. This supplementation positively improved the ratio between total Coli and Lactobacilli; this presumably contributed to the maintenance of a good health of the subjects.

In the similar trial 51 B executed by this Department in 1999, a positive effect on the growth performance and pH was observed.

The absence of a evident effect of Turval B0399 in this parameter of the present trial, could be attributed to differences in the base diet, that could have affect also the presence of other bacteria species not cultivated or not cultivable.

This could have consequently contributed to maintain a similar performances results between the different diets.

In the case of the dietary overdose (tolerance test) with Turval B0399 the growth performance of weaned piglets was not influenced, but the health was impaired, at least in the 4 subjects that died for intestinal pathologies. In the other subjects the pH in the gut were nod modified.

The balance between Coli and Lactobacilli was sensibly improved. Presumably this observation would not have been found in the subjects that died, however the data do not permit to identify the reasons why these four subjects performed differently.

 

It is interesting to note that in 1/3 (Diet T1) to 2/3 (Diet T2: tolerance test) of the subjects fed the additive, the Turval B0399 was recovered still cultivable in the colon content. Furthermore the data show that apparently no contamination was present between the different groups.