Pakistan Journal of Science (Vol. 73 No.

1 March, 2021)

IMMUNO-MODULATORY EFFECTS OF LACTOBACILLUS IN SALMONELLA

GALLINARUM INFECTED BROILER CHICKS
A. Abbas1, F. Rizvi1, A. Abbas2, S. Hussain2,S. Ali2, S. Ali3, R. Rafique3, A. W. Manzoor2, H. M. Waqar2, R. Akram4,
M.Shaukat5, H. Shaukat6 and R. Rafique7
1
University of Agriculture Faisalabad, Pakistan
2
Veterinary Research Institute, Zarrar Shaheed Road, Lahore Cantt. Pakistan
3
Foot and Mouth Disease Research Centre, Zarrar Shaheed Road, Lahore Cantt. Paksitan
4
University of Veterinary & Animal Sciences Lahore, Pakistan
5
School of Biological Sciences, University of the Punjab, Quaid-i-Azam Campus Lahore-54590, Pakistan
6
Department of Zoology, University of the Punjab, Lahore, Pakistan
7
PMAS Arid Agriculture University Rawalpindi, Pakistan

ABSTRACT: Lactobacillus is a direct fed microbial which helps to protect organisms against
pathogens by discouraging their colonization in intestine. This project had been designed to evaluate
the effect of Lactobacillus in broiler chicken experimentally infected with S. gallinarum. A total of 100
day old chicks divided into 4 equal groups (A, B, C & D) comprising 25 chicks in each group. Birds of
groups B and D were infected with S. gallinarum orally at day 19th of age. Group A was kept as control
negative and group C and D were supplemented with Lactobacillus. Disease combat efficiency in each
group was determined by monitoring weight gains, morbidity & mortality rates, immune parameters
and histopathological findings. Analysis of variance (ANOVA) technique was used to compare results
by using MSTAT C statistical software. The results revealed that morbidity and mortality rates were
more in group B and significantly low in group C (Lactobacillus supplemented) and D (Treatment
group). The antibody response was found to be highest in group C followed by group D. Body weight
gains and relative organ weight gains were also higher in treated groups with maximum in the group C
followed by group D. Histopathology studies revealed that treated groups showed less lesions and
untreated infected group showed all classical lesions of the disease. The study concluded that
Lactobacillus can be very efficient as probiotic for prophylactic immune-stimulation to replace
antibiotics being used irrationally as prophylaxis.
Key words: Immuno-modulatory, Lactobacillus, Probiotics, Salmonella gallinarum, Broiler chicks.
(Received 05.11.2020 Accepted 30.11.2020)

INTRODUCTION diarrhea, dehydration and in layers it leads to sudden drop

in egg production (Dey et al., 2016).
Poultry sector is one of the most vibrant sub- The accessibility of antibiotics to be used for
sectors of livestock. The current investment in poultry treatment of infective diseases has considerably improved
industry is more than Rs.700 billion. This industry is the human health and animal welfare. The massive
growing at the rate of 8% per annum over the last few misuse of antibiotics causes the development of
years. Pakistan is the 11th largest producer in the world antimicrobial resistance in commensal and pathogenic
with 1.163 million broilers annually and also provides bacteria (Carattoli 2008, Depoorter et al., 2012).
employment to more than 1.5 million people Antibiotic residues in the poultry meat and drug
(Anonymous, 2019-2020). A person requires 102.7 grams resistance are basic threats for antibiotic use for curative
of protein on daily basis but almost 66% of Pakistani and preventive purposes. Now it is much needed to use
population is deficient in animal protein source alternative of antibiotics, not as prophylactic measure but
(Abedullah, 2007). Poultry industry is vulnerable to many also for therapeutic purpose. Use of prebiotics, probiotics,
infectious and non-infectious diseases which are the main organic acids and plant extracts can overcome this
obstacle in the development of poultry industry. Among problem (Griggs et al., 2005). Probiotics can improve
communicable diseases, Fowl Typhoid is an important growth of beneficial bacteria by inhibiting the
disease which is caused by S. enteric serovar Gallinarum multiplication of pathogenic organisms (Li et al., 2018).
(S. gallinarum). S. gallinarum is facultative anaerobe, Lactobacillus supports intestinal epithelial
non-motile, rod shaped and gram-negative bacterium tissues of the host against pathogenic bacteria
which is host specific in nature. The most prominent (Klaenhammer et al., 2000). Lactobacillus is a micro-
clinical signs in Fowl Typhoid are sulpher colored aerophilic anaerobe and gram positive bacteria which has

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 Pakistan Journal of Science (Vol. 73 No. 1 March, 2021)

ability to proliferate at low pH. It has beneficial effects the dose rate of @109 Colony Forming Units (CFU) and
on gut of animal by competitive exclusion of pathogenic group D infected with Lactobacillus and S. gallinarum at
microbes and by producing lactic acid. It can also dose rate of @109 CFU and 108 CFU respectively on 19th
promote the production of antibodies by stimulating host day of experiment through oral route. Afterwards
humoral immune response and can trigger T-cells of infection was induced, seven birds were culled weekly
immune system (Talebi et al., 2008). In poultry, from each group and blood was collected for
Lactobacillus heat killed strains like LAH7, LAP5, LAF1 hematological evaluation. For histopathological studies
have the ability to adhere with GIT epithelium and organs were preserved in 10% buffered formalin solution
protect the host from infectious micro-organism. In return (Bancroft and Gamble., 2007).
it inhibits the growth of Salmonella and E. coli (Jin et al., Broiler birds were kept under observation for 6
1998). Other strain such as LF33 also has the ability to weeks. Clinical signs, gross lesions and organs weight
adhere with intestinal cells and inhibit the activity of E. were observed. At 21st, 28th, 35th and 42nd days, 6 birds
coli, Staphylococcus aureus and S. typhimurium. Thus the were slaughtered from each group and organs having
issue of the antimicrobial resistance and antibiotic lesions were recorded. For histopathological studies
residues in the poultry can be minimized by the use of organs including spleen, bursa of fabricius, thymus, liver
probiotics in poultry. and heart were preserved in 10% buffered formalin
The current project was designed to use (Bancroft and Gamble., 2007).
Lactobacillus as probiotic in poultry feed to determine Immunological parameters including IgG, IgM
the efficacy of Lactobacillus against S. gallinarum and IgA were measured by commercial ELISA kit.
infection, immunomodulatory action of Lactobacillus EDTA added blood samples were collected from each
supplementation and pathological changes in broiler group, hematological parameters including total
chicks. erythrocyte counts (TEC), total leukocyte counts (TLC),
packed cell volume (PCV) and hemoglobin concentration
MATERIALS AND METHODS (Hb) were estimated.
Statistical analysis: Analysis of variance technique and
Lactobacillus samples were collected by mixing means were compared by DMR, ANOVA test by using
1 ml of yogurt with 10 ml of normal saline. For its MSTAT C statistical software.
propagation, MRS (de Man, Rogosa, Sharpe agar) media
was used which is selective for Lactobacillus. For MRS
RESULTS
agar 62 grams of MRS powder was mixed in one liter of
water. Ten ml of this mixture was poured in petri dish.
Isolation and Identification of Bacteria: The colonies
MRS broth was prepared by mixing 52 grams of MRS
of Lactobacillus were observed which were raised,
powder in one liter water in Pyrex flask. Ten ml of this
whitish in color and had sand like appearance.
mixture was poured in test tube. Media was sterilized at
Microscopically rod shaped, purple colored and gram
121oC for 20 minutes at 15lbs atm. Yogurt mixture was
positive bacteria were seen (Fig. 1).
streaked on four plates from every batch and kept at 37 oC
and 25oC respectively. After 72 hours plates were Feed intake: Feed intake was significantly increased in
examined and absence of growth showed sterility the birds of control negative, treatment group and
(Cruickshank, 1975). supplemented with Lactobacillus alone as compared to
For identification of Lactobacillus bacterial the birds of control positive group (Table 1).
smear was prepared by applying a drop of bacterial
suspension on a glass slide. It was flooded with 1.0% of Morbidity and Mortality Percentage: Clinical signs
crystal violet followed by dipping in 5% NaHCO3 like depression, diarrhea, ruffled feathers and anemia
were observed in infected and treated groups (Fig. 2).
solution for 3 minutes. Gram iodine solution was poured
Percentage of clinical signs were higher in infected group
on slide for 2 minutes followed by decolorisation with
as compared to the treatment. Group A was kept as
acid alcohol for 10 seconds. Glass slide was tilted and
counter stain i.e. safranin was used for 30 seconds, slide control group, no infection was given and it was kept in a
was washed, air dried and observed under 40X and 100X separate room. Morbidity and mortality were not seen in
this group. Chicks of group B showed highest morbidity
oil immersion lense.
(52%) and mortality (48%) due to S. gallinarum infection
A total of 100 day old chicks were purchased
as compared to all other groups. No morbidity and
from hatchery and divided into four groups (A, B, C &
mortality were observed in group C. In the birds of group
D) comprising 25 chicks in each group. All chicks were
vaccinated against ND, IBV, IBD (D78, 228E) and HPS. D 28% morbidity and 24 % mortality were observed
Feed and water were provided ad libitum. The birds in (Table 2).
group A were kept as negative control, group B as
positive control, group C infected with Lactobacillus at

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Hematological Parameters: Total erythrocytes count, levels were found to be higher in Lactobacillus treated
Hemoglobin (Hb) concentration and Packed cell volume groups rather than control groups (Table 9).
percentage were significantly decreased in infected birds
Gross Lesions: Seven birds from group A were culled on
as compared to other treated groups. There was non-
every week. Birds of this group were non infected and
significant relationship between control negative (group
non-supplemented. Thus no postmortem lesions were
A) and supplemented group (group C). Total leukocytic
seen in this group. Postmortem was organized for
count was significantly increased in infected group as
mortality birds to examine lesions of fowl typhoid. Group
compared to other treated groups and there was no
B showed severe lesions in heart (pericarditis), liver
significant difference between control negative group A
(perihepatitis) and spleen (splenomegaly). In group C
and Lactobacillus supplemented group C (Table 3).
supplemented with Lactobacillus, mild postmortem
Body Weight and Organs Weight Parameters: Body lesions were seen. In this group only pericarditis was
weight gain was significantly decreased in infected group seen. In group D, pericarditis and perihepatitis were seen
as compared to treated group but there was non- during postmortem of chickens at 7th, 14th and 21st day
significant relationship between control negative and post infection (Fig. 3).
supplemented group. Absolute and relative weight of
Histopathology: Birds of group B and D exhibited
spleen (Table 4), thymus (Table 5), bursa (Table 6),
microscopic changes on histopathological examination.
kidney (Table 7) and liver (Table 8) were significantly
Lymphocyte proliferation and degenerative changes were
increased in infected group (control positive) as
seen under microscope. Hemorrhagic spots and
compared to the treated group (group D) while non-
congestion were also seen in the infected birds. Birds of
significant relation between control negative and
control positive and treatment plus infected groups
supplemented group were recorded.
showed microscopic alterations upon histopathological
Immunomodulatory Effects: Lactobacillus was tested examination in lungs. Marked increase in congestion due
as a probiotics with possible immunomodulatory effects to rupture of inter alveolar septa was observed (Fig. 4).
to combat S. gallinarum infection. The IgG and IgA

Table 1. Feed intake (gm) of broiler birds supplemented with Lactobacillus and infected with S. gallinarum.

Weeks Group A Group B Group C Group D

1st 18.60±6.83a 17.91±6.05a 18.31±6.92a 18.66±6.88a
2nd 54.56±13.55a 52.2±10.05a 56.59±18.91a 52.66±14.55a
3rd 105.48±10.84bc 92.16±6.88c 122.27±8.44a 113.51±14.88ab
4th 152.88±15.20a 95.01±10.98b 159.14±18.21a 158.66±11.05a
5th 177.80±4.29b 120.10±3.05c 192.45±7.40a 188.67±6.97a
Mean ± SE having similar alphabets in a row are statistically non-significant (P> 0.05)
A= No Infection (Control –Ve)
B= Infection with S. gallinarum (Control +Ve)
C= Supplemented with Lactobacillus
D= Infected with S. gallinarum and Supplemented with Lactobacillus

Table 2. Percentage morbidity and mortality in experimental groups.

Morbidity Mortality
Group
No. % No %
A - - - -
B 13 52 12 48
C - - - -
D 7 28 6 24
A= No Infection (Control –Ve)
B= Infection with S. gallinarum (Control +Ve)
C= Supplemented with Lactobacillus
D= Infected with S. gallinarum and Supplemented with Lactobacillus

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Table 3. Red blood cells count (106), Total leukocyte Count (103), Pack cell volume (%) and Hemoglobin concentration (g/dl) in S. gallinarum infected
broiler chicks supplemented with Lactobacillus

Days post Hematology Groups

Infection Parameters A B C D
RBC 3.12±0.011bc 2.50±0.098e 3.38±0.075a 2.90±0.026d
TLC 22.55±0.074gh 32.85±0.139a 21.85±0.711h 28.66±0.186b
7
PCV 35.99±0.19bc 26.79±0.49g 37.67±0.33a 33.21±0.26d
Hb 11.53±0.081def 8.97±0.115g 12.56±0.149ab 11.45±0.185ef
RBC 3.08±0.014bc 2.33±0.043f 3.17±0.049b 2.90±0.025d
TLC 25.47±0.158e 29.17±0.115b 24.67±0.309ef 25.81±0.273de
14
PCV 32.63±0.10de 23.76±0.13h 36.81±0.41b 31.20±0.30f
Hb 12.16±0.092bc 7.986±0.118h 12.68±0.109a 11.34±0.243f
RBC 3.01±0.018cd 2.32±0.041f 3.05±0.031bc 2.88±0.043d
TLC 23.81±0.106fg 28.20±0.141bc 22.81±0.346gh 27.03±1.501cd
21
PCV 30.54±0.10f 22.97±0.09h 32.11±0.35e 35.75±0.45c
Hb 11.66±0.108def 7.13±0.163i 11.88±0.142cd 11.86±0.184cde
Mean ± SE having similar alphabets in a row are statistically non-significant (P> 0.05)
A= No Infection (Control–Ve)
B= Infection with S. gallinarum (Control +Ve)
C= Supplemented with Lactobacillus
D= Challenged with S. gallinarum and challenged with Lactobacillus

Table 4. Absolute and relative weight (gm) of Spleen of S. gallinarum infected broiler chicks supplemented with Lactobacillus.

Days Weight (gm)

Post A B C D
Infection Absolute Relative Absolute Relative Absolute Relative Absolute Relative
7 1.40±0.216 fg 0.193±0.011bc 2.84±0.453 b 0.413±0.026 a 1.56±0.127ef 0.177±0.007 bcd 11.69±0.090 de 0.194±0.006 bc
14 1.49±0.135 efg 0.121±0.006cd 2.96±0.270 b 0.264±0.015 b 1.37±0.364fg 0.097±0.010 d 1.21±0.168 g 0.097±0.007 d
21 1.93±0.206cd 0.097±0.006d 3.69±0.406 a 0.219±0.018b 2.057±0.151 c 0.094±0.003 d 2.17±0.180 c 0.109±0.006 cd
Mean ± SE having similar alphabets in a row are statistically non-significant (P> 0.05)
A= No Infection (Control –Ve)
B= Infection with S. gallinarum (Control +Ve)
C= Supplemented with Lactobacillus109 CFU orally
D= Infected with S. gallinarum and Supplemented with Lactobacillus

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Table 5. Absolute and relative weight of Thymus (gm) in S. gallinarum infected broiler chicks supplemented with Lactobacillus.

Days Weight (gm)

Post A B C D
Infection Absolute Relative Absolute Relative Absolute Relative Absolute Relative
7 1.94±0.270 cd 0.269±0.017 b 1.64±0.172 f 0.243±0.009 b 2.29±0.241 a 0.260±0.011 b 2.34±0.215 a 0.271±0.007 b
14 2.03±0.099 bcd 0.164±0.009 b 1.80±0.064 def 0.163±0.012 b 1.89±0.248 de 0.131±0.005 b 1.69±0.177 ef 0.133±0.005 c
21 2.16±0.223 abc 0.109±0.006 b 1.40±0.216 g 0.083±0.007 b 2.35±0.418 a 0.110±0.008 a 2.20±0.141 ab 0.111±0.008 a
Mean ± SE having similar alphabets in a row are statistically non-significant (P> 0.05)
A= No Infection (Control –Ve)
B= Infected with S. gallinarum (Control +Ve)
C= Supplemented with Lactobacillus109 CFU
D= Infected with S. gallinarum and Supplemented with Lactobacillus

Table 6. Absolute and relative weight of Bursa (gm) in S. gallinarum infected broiler chicks supplemented with Lactobacillus

Days Weight (gm)

Post A B C D
Infection Absolute Relative Absolute Relative Absolute Relative Absolute Relative
7 1.84±0.140 bc 0.253±0.00 8b 1.46±0.151 e 0.213±0.010 b 1.787±0.205 bc 0.203±0.00 8b 1.74±0.151 cd 0.197±0.005 b
14 1.91±0.195 bc 0.154±0.005 b 1.54±0.190 de 0.139±0.012 b 1.84±0.162 bc 0.129±0.003 b 1.56±0.207 de 0.126±0.011 c
21 2.16±0.223 a 0.109±0.006 b 1.40±0.216 e 0.083±0.007 b 2.15 ±0.127 a 0.099±0.001 a 1.97±0.302 ab 0.099±0.001 a
Mean ± SE having similar alphabets in a row are statistically non-significant (P> 0.05)
A= No Infection (Control –Ve)
B= Infected with S. gallinarum (Control +Ve)
C= Supplemented with Lactobacillus109 CFU
D= Infected with S. gallinarum and Supplemented with Lactobacillus

Table 7. Absolute and relative weight of Kidney (gm) in S. gallinarum infected broiler chicks supplemented with Lactobacillus

Days Weight (gm)

Post A B C D
Infection Absolute Relative Absolute Relative Absolute Relative Absolute Relative
7 7.87±0.515 b 1.090±0.030 abc 9.21±0.438 b 1.351±0.029 ab 8.23±0.457 b 0.937±0.027 abc 8.26±0.680 b 0.951±0.015 abc
14 8.70±0.480 b 0.709±0.034 bc 10.53±0.559 b 0.941±0.053 abc 7.47±0.640 b 0.521±0.017 bc 22.79±34.958 a 1.757±0.997 a
bc
21 14.14±2.031 ab
0.716±0.024 16.91±1.783 ab
0.990±0.055 abc 9.90±0.305 b 0.457±0.004 c 11.27±0.846 b 0.566±0.038 bc
Mean ± SE having similar alphabets in a row are statistically non-significant (P> 0.05)
A= No Infection (Control –Ve)
B= Infected with S. gallinarum (Control +Ve)
C= Supplemented with Lactobacillus109 CFU
D= Infected with S. gallinarum and Supplemented with Lactobacillus

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Table 8. Absolute and relative weight of liver (gm) in S. gallinarum infected broiler chicks supplemented with Lactobacillus.

Days Weight (gm)

Post A B C D
Infection Absolute Relative Absolute Relative Absolute Relative Absolute Relative
7 35.44±0.736def 4.904±0.141b 46.26±1.310c 6.783±0.181a 31.98±0.337f 3.646±0.094bcd 33.17±0.170ef 3.830±0.104bc
14 36.67 ±0.733de 2.971±0.114cde 53.00±2.93b 4.711±0.273b 38.84±0.563d 2.719±0.079cde 35.53±1.705def 2.800±0.076cde
21 50.59±1.413bc 2.569±0.049cde 75.86 ±3.865a 4.440±0.289b 48.43±0.95bc 2.237±0.034e 47.17±0.922c 2.354±0.100de
Mean ± SE having similar alphabets in a row are statistically non-significant (P> 0.05)
A= No Infection (Control –Ve)
B= Infection with S. gallinarum (Control +Ve)
C= Supplemented with Lactobacillus109 CFU
D= Infected with S. gallinarum and Supplemented with Lactobacillus

Table 9. Immunoglobulin A (IgA) and Immunoglobin G (IgG) in broiler chicks supplemented with Lactobacillus and infected with S. gallinarum.

Days Group A Group B Group C Group D

Post
IgA IgG IgA IgG IgA IgG IgA IgG
Infection
ef cd g e ab bc cd
7th 0.39±0.007 4.25±0.056 0.28±0.007 2.89±0.095 0.49±0.08 5.18±0.048 0.35±0.007 3.34±0.052de
14th 0.49±0.006cd 5.28±0.066bc 0.035±0.005ef 3.26±0.034de 0.56±0.008abc 6.19±0.082ab 0.48±0.010ab 4.22±0.048cd
21th 0.60±0.008ab 6.39±0.105ab 0.46±0.007de 4.28±0.114cd 0.64±0.007cd 7.14±0.084a 0.54±0.013bcd 5.48±0.274bc
Mean ± SE having similar alphabets in a row are statistically non-significant (P> 0.05)
A= No Infection (Control –Ve)
B= Infected with S. gallinarum (Control +Ve)
C= Supplemented with Lactobacillus
D=Challenged with S. gallinarum and supplemented with Lactobacillus

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White opaque colored colonies of Lactobacillus on Purple stained Lactobacillus by Gram staining
MRS media
Figure1. Isolation and identification of Lactobacillus .

Figure 2. Depressed birds with ruffled feathers from treatment group and healthy group.

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Inflamed liver with necrotic foci in control positive group No inflammation seen in liver of treated group

Difference in size of spleen in control positive and control Difference of liver size in control positive and
negative group treatment groups
Figure 3. Lesions seen in different organs.

Photomicrograph of liver showing diffuse hemorrhage and Photomicrograph of lung tissue showing
inflammatory cellular infiltration inflammatory cellular infiltration.
Figure 4. Histopathological changes in liver and lung.

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DISCUSSION There was decrease in the morbidity and

mortality rates in all of Lactobacillus treated groups as
Poultry industry is one of the largest agro based compared to the control group. Same findings were seen
sector of Pakistan established in 1962. A total of 45-50 by Khan et al. (2010) who reported about the
percent meat demand is being fulfilled by poultry. supplementation of 40 mg/kg of Lactobacillus against S.
Therefore it is necessary to develop poultry industry on gallinarum infected broiler chicks and observed decrease
commercial basis to meet the baseline requirement of in mortality rate.
meat and eggs. Poultry industry is exposed to many Absolute weight of liver, spleen and heart were
infectious and non-infectious diseases which are the main significantly decreased in treated groups which were
obstacle in the development of poultry industry (Hussain supplemented by Lactobacillus as compared to other non-
et al., 2015). Among infectious diseases fowl typhoid is a treated groups. Similar findings were reported by Huff et
major disease caused by Salmonella gallinarum and it al. (2006) that after 25 days of Lactobacillus
causes lesions on multiple visceral organs. The disease is supplementation, absolute weight of the heart, spleen and
responsible for high morbidity and mortality rates in liver were decreased against S. gallinarum infected
chicken flocks worldwide (Riaz and Aslam, 2016). broiler birds.
Recently farmers are using antibiotics Birds which were supplemented by
irrationally to overcome their fear of loss of their chicken Lactobacillus and challenged by S. gallinarum, WBCs
flocks. Now the concept of antibiotic free meat has were increased as compared to the control negative
obtained strength and the demand for antibiotic free group. Same results were observed by Dong et al. (2013)
chicken has increased. Therefore stress is being given on who reported that WBCs count was significantly
finding alternatives to antibiotics and major breakthrough increased from 4.08 to 4.55 after the Lactobacillus
are prebiotics and probiotics (Patterson and Burkholder, supplementation. Hemoglobin concentration (Hb) was
2003). Probiotics provide a rational alternative to increased significantly in Lactobacillus supplemented
antibiotic use. Among probiotics, the Lactobacillus birds. Same results were reported by Sharaity et al.
species are most common, found in feed such as yogurt, (2017) who observed increase in hemoglobin
cheese, beer and fermented food etc. Many probiotics concentration in probiotic supplemented birds.
along with their feasibility are safe and effective for There were severe hepatitis, nephritis and
treatment. Tellez et al. (2012) reported Lactobacillus to splenomegaly in broiler chicks of control positive
be very safe by not causing super-infections and also (infected with S. gallinarum) group while in group A
found to elicit up to 201 genes for immune stimulation (non-infected) broiler chicks no lesions were observed.
response. These findings were also in agreement with the results of
The objective of this trial was to evaluate the Tonu et al. (2011). Lactobacillus was tested as a
effects of Lactobacillus supplementation on weight gain, probiotics with possible immunomodulatory effects to
pathology, hematology and immune response of broiler combat S. gallinarum infection. The IgG and IgA levels
chicks infected with field isolate of the S. gallinarum. were found to be higher in Lactobacillus treated groups
Feed intake and body weight of Lactobacillus rather than control groups. Gill et al. (2001) found similar
supplemented groups were significantly increased as results and reported immunomodulatory effects of
compared to S. gallinarum infected groups. Probiotic probiotic (Lactobacillus) against E. coli infection given to
(Lactobacillus) has beneficial effect on weight gain of mice.
broiler chickens and similar findings were recorded by Conclusion: The present study concludes that
AN et al. (2008). The results are also in accordance with supplementation of Lactobacillus @109 CFU has positive
the findings of Marota et al. (2019) who illustrated the effects on the growth performance, decreased morbidity
beneficial effects of Probiotics on public health. and mortality rates of broiler chicks during fowl typhoid.
The infected groups showed clinical signs At the same time Lactobacillus also helps to increase the
including off feed, ruffled feathers, depression, lethargy, immunoglobulin level in blood and improves the health
lameness and being unable to stand. While in group D of birds.
(treatment group), dull physical appearance and leg
weakness were prominent signs. No clinical signs were
observed in group A (control negative) and group C (only
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