Printer Friendly

SCREENING OF LACTIC ACID BACTERIA FOR THEIR USE AS BUFFALO PROBIOTIC.

Byline: S. Khan, A-Ud-Din, G. M. Ali, S. I. Khan, I. Arif, M. N. Riaz and S. Ghazanfar

Keywords: Lactobacillus, Acid tolerance, Lactic acid bacteria, Breed specific probiotic, Phylogenetic analysis.

INTRODUCTION

Dairy products industry is one of the fastest growing industries of the world. Previous studies have shown that 99% of milk production is coming from ruminants. The lactating animals are highly dependent on their gut microbial communities to fulfil their nutritional needs for maintaining body homeostasis, milk production and growth. The diverse and multifunctional animal microbial flora present in gastrointestinal tract may play a significant function in high or low milk production. Numerous techniques have been employed to enhance animals' productivity by manipulating GIT microbial communities of dairy animals. The use of antibiotics to improve dairy animal's productivity is associated with an increasing emergence of antibiotic resistance in microbiota (Zeineldin et al., 2008). This emergence led to ban on the use of antibiotics as a feed additive in many countries (Lee and Salminen, 2009).

Livestock sector is seeking alternative to antibiotics which could safely fulfil the demands of the local markets. The use of friendly bacterial feed supplements (probiotics) is one of the recommended methods to improve the balance of GIT microbes (GU and Roberts 2019). Probiotics are live microbes that benefit the host by maintaining the GIT microbial balance (Fuller, 1989). Further, till date they have been discovered non-pathogenic microbes that occur naturally in the tract of ruminants. Various microorganisms have been used as probiotics (Di-Gioia et al., 2018). Probiotics bacteria give positive impact in young animals, whereas probiotic yeasts are found to be effective in lactating animals (Markowiak and Slizewska, 2018).

The most common ruminant probiotic products available in market are yeast probiotics i.e. Saccharomyces cerevisiae and bacterial probiotics i.e. lactic acid bacteria (LAB), Bifidobacterium, and some Bacillus (Kvan et al., 2018; Grochowska et al., 2012). Among LAB, Lactobacillus acidophilus, L. plantarum, L. casei, L. Lactis and Enterococcus faecium are found as one of the best candidates for probiotics. It has been reported that probiotic feed supplements are highly responsive to geographical location of the host, age, diet and breed Taylor et al., (2007). Probiotic effectiveness is highly dependent on the host and probiotic strain for maximum colonization Collado et al., (2009). The host origin of commercially available probiotic strains is not specific. Therefore, LAB strains fed to buffalo should be isolated from the same animal source or niche for obtaining maximum benefits in terms of meat and milk production (Shakira et al., 2018).

The present study was designed with the objective to isolate and molecular identification of the autochthonous LAB strains present in buffalo gut for their potential use in buffalo feed as buffalo probiotic.

MATERIALS AND METHODS

Sampling from Nilli Ravi buffalo: Fresh fecal samples (10 g) from 18 lactating dairy buffalo (Nilli Ravi) were taken from deep rectum using the sterile gloves from Livestock Research Station, National Agricultural Centre (NARC), Islamabad, and district Buner, Khyber Pukhtunkhwa Pakistan. The samples were taken early in the morning in the sterile containers (Deltalab, Spain) and immediately transported to the laboratory for microbiological study.

Microbiological analysis: For microbial profiling, 5 g fecal samples from each group was homogenized in 40 ml saline-peptone water in a falcon tube using vortex mixture for 2 min. The serial dilutions were then prepared in saline (NaCl 0.9 w/v). Microbial suspension were plated on MRS media (Oxoid, UK) and incubated for 24 to 48 h at 37 AdegC under anaerobic condition (BD BBL(tm) GasPak(tm)) for total lactic acid bacterial species (Lactobacillus, Enterococcus, Lactococcus, and Bifidobacteria)

Isolation of buffalo origin bacterial strains: Methods of bacterial isolation were adopted to obtain the LAB strains on DeMan, Rogosa and Sharpe agar, (MRS) (Oxoid, UK) media. Buffalo samples after serial dilution in PBS buffer were spread on MRS media plates. The plates were aerobically incubated at 37 AdegC for 48 hours (Di-Gioia et al., 2016). Morphological characterisation of bacterial strains was done by using Bergey's manual (1962).

Selection of buffalo origin presumptive probiotic strains: 20 catalase negative and gram positive bacterial strains were inoculated in MRS medium containing 0.01 lactic acid. The concentration was prepared according to lactic acid concentrations in the buffalo rumen (Van Nieuwenhove et al., 2007). The bacterial isolates were incubated for 170 h at 28AdegC, the bacterial colony size was noted every 24 h during that incubation period (Table 8). The strains that showed maximum cellular viable results were selected for antimicrobial test. For this the bacterial isolates were inoculated at 37AdegC in shaking incubator on MRS media. Five pathogenic bacterial strains L. monocytogenes, S. aureus, E.coli, B.cereus and P.aeroginosa were selected to test antimicrobial effect by well diffusion agar assay method (Naeem et al., 2018).

Identification of buffalo origin probiotic strains by 16S rRNA gene sequence

Extraction of Genomic DNA: Genomic DNA extraction from the pure bacterial isolates was done by using the methods given by Shakira et al., (2018). It involved suspension of purified bacterial colonies of single strain in 20uL of Tris EDTA buffer in micro-PCR strips. The bacterial strain colonies were mixed gently and heating the PCR machine at 95 AdegC (10 min). The mixture was centrifugate at 6000 rpm (2-3 min). The supernatant obtained by discarding the pellet was used as a DNA template for the amplification of 16S rRNA gene.

PCR-based buffalo origin bacterial strains identification: Takara Pre-Mix Ex-Taq was used to amplify the 16S rRNA gene of the presumptive LAB strains by using the method given by Shakira et al., (2016). The sequencing for the 16S gene of the amplified PCR products was conducted using commercial sequencing service of Macrogen Inc Korea.

Determination of animal probiotic characterization

Acid tolerance: To determine low pH the tolerance of LAB strains, a suspension on MRS broth was prepared. The absorption was adjusted at 600 nm wavelength. The suspension was centrifuged and suspended in phosphate solubilizing bacteria (PSB). Different pH suspension were made and incubated for 3 hours and later on, aliquot inoculated in MRS broth. Strains were serially diluted and inoculated on MRS agar for measure cfu/ml (Hassan zadazar et al., 2012).

Antibiotic resistance: Disc diffusion method was used for determination of the antibiotic resistance (Olatunde et al., 2018). LAB strains were anaerobically grown at 30AoC in MRS broth for 24 h. Antibiotic discs were placed on overlaid prepared plates and incubated at 30AoC for 24 h anaerobically.

Hemolytic activity: The bacterial strains inoculated on MRS broth and then streaked lined on blood agar dispensed with sterile 5% defibrinated blood incubated at 37AdegC for 48 hours. Results were noted for a deep hemolysis zone around the bacterial colonies (Halder et al., 2017).

Enzymatic Activity: The proteolytic activity of LAB was determined by autoclaving, skim milk agar (SMA) plates. 10 g skim milk powder was added to autoclaved SMA plates. The 24 hours LAB isolate were streaked on the prepared plates and aerobically indicated at 37 o C for 48 h. For determination of amylolytic activity amylase media plates were prepared. The freshly grown inoculum was streaked on the plates and 37 o C (Latorre et al., 2016).

-galactosidase activity: The [beta]-galactosidase activity of LAB was determined by method given by Chen et al., (2008).

Bile salt tolerance and cholesterol assimilation: Bile salt tolerance and cell surface hydrophobicity of the LAB were determined by the method given by Del Re et al., (2000).

Statistical Analysis: All experimental tests were performed in triplicates. Results are presented as mean with standard deviation. The statistical analysis was done by using the statistics (version 8.1) software. The significant difference between the means was assessed by using Tukey's test. (Pa$? 0.05).

Table 1. Morphological and biochemical characterization of LAB strains as candidate for buffalo probiotics.

Characterization###Enterococcus###Pediococcus###Enterococcus###Lactobacillus###Pediococcus###Enterococcus

###lactis###pentosaceus###faecalis###fermentum###acidilactici###ratti

Shape###Cocci###Cocci###Cocci###Rod shape###Cocci,###Cocci

###occurs in

###tetrads

Gram Staining###+###+###+###+###+###+

Catalase###-###-###-###-###-###-

Oxidase###-###-###-###-###-###-

Gas from###+###-###-###+###-###-

Glucose

Fermentation###Homo###Homo###Homo###Hetero###Homo###homo

Hemolysis###GAMMA###GAMMA###GAMMA###GAMMA###GAMMA###GAMMA

###Growth at temperature (OC)

04###-###-###-###-###-###-

10###+###-###+###-###-###+

37###++###++###++###+###++###++

45###+###+###+###++###++###+

50###-###+###-###-###+###-

Table 2. The antibacterial activity of LAB strains as candidate for buffalo probiotics against five pathogens and their zones diameter (mm).

NMCC###Test Pathogen

Strains###E. coli###Pseudomonas###Staphylococcus###Listeria###Bacillus

###aeruginosa###aureus###monocytogenes###Cereus

NMCC-PI###-###-###+###+++###+

NMCC- PT###-###+###+###+++###-

NMCC-P3n###-###+###-###++###+++

NMCC-P7###++###+###++###+###+

NMCC-P8###++###+###+++###+++###++

NMCC-F###-###+###++###++###++

Table 3. 16S rRNA based gene analysis of LAB strains as candidate for buffalo probiotics.

NMCC Strain###Bacterial strain###Length of 16S###Accession###Bacterial Taxonomy###Similarities

###name (genus)###r RNA (ntds)###number

NMCC-PI###Enterococcus###1028###MK007471###Enterococcus lactis###98.66

NMCC- PT###Pediococcus###1119###MK014295###Pediococcus pentosaceus###97.74

NMCC-P3n###Enterococcus###1196###MK007347###Enterococcus faecalis###95.32

NMCC-P7###Lactobacillus###1242###MK007323###Lactobacillus fermentum###94.45

NMCC-P8###Pediococcus###1176###MK007346###Pediococcus acidilactici###96.58

NMCC-F###Enterococcus###1190###MK014298###Enterococcus ratti###97.67

Table 4. Different pH (2, 4, 5, and 6) effects on survival of LAB strains as candidate for buffalo probiotics.

NMCC Strain###pH (2.0)###pH (4.0)###pH (5.0)###pH (6.0)

###log CFU/ml

NMCC-PI###6.47+-0.04a###3.71+-0.02a###5.32+-0.04a###5.65+-0.06a

NMCC- PT###6.89+-0.03a###3.54+-0.06ab###5.45+-0.06c###6.55+-0.07a

NMCC-P3n###6.34+-0.06a###3.12+-0.08b###5.35+-0.08a###6.34+-0.04a

NMCC-P7###7.86+-0.08a###6.36+-0.06b###5.29 +-0.03c###6.24+-0.088ab

NMCC-P5n###7.01+-0.11a###6.55+-0.04b###6.23+-0.07###6.54+-0.09

NMCC-F###6.34+-0.09a###3.12+-0.05a###5.35+-0.01a###6.33+-0.02a

Table 5. Antibiotic resistance profiles of LAB strains as candidate for buffalo probiotics against commonly used antibacterial compounds.

NMCC###Strea###Ciprb###Vanc###Metrd###Ampie###Chloraf###Kanag###Eryth###Peni###Tetj

Strain###(10ug)###(20ug)###(30ug)###(10ug)###(5ug)###(30ug)###(30ug)###(15ug)###(10ug)###(30ug)

NMCC-PI###S###S###R###R###R###S###S###R###R###R

NMCC- PT###R###R###R###R###R###I###R###S###R###S

NMCC-###S###I###S###R###R###S###S###R###S###R

P3n

NMCC-P7###R###R###R###R###I###S###R###S###R###R

NMCC-P8###R###R###R###R###S###I###R###S###R###R

NMCC-F###S###S###S###R###S###R###S###R###R###R

Table 6. Enzymatic potential of LAB strains as candidate for buffalo probiotics.

NMCC Strain###Lipolytic Activity###Proteolytic Activity###Amylolytic Activity

NMCC-PI###+###+###-

NMCC- PT###++###+###+

NMCC-P3n###++###+###-

NMCC-P7###+###++###++

NMCC-P8###++###++###+

NMCC-F###+###-###-

Table 7. In-vitro tests of LAB strains as candidate for buffalo probiotics (n=3).

NMCC Strain###Cholesterol assimilation (%)###[beta]-galactosidase activity

NMCC-PI###37.34+-1.76a###410.12+-25a

NMCC- PT###46.98+-1.34b###290+-41b

NMCC-P3n###37.13+-1.65c###-

NMCC-P7###41.34+-1.54d###347 +- 47c

NMCC-P8###44.35+-1.45e###276+-29d

NMCC-F###36.55+-0.39a###421+-54

Table 8. LAB strain shows cell viability, temperature and lactic acid assimilation.

Bcateria1###Cell viability in rumen fluid###Temperature (AdegC)###Lactic acid assimilation

###(Log10 cfu, mL-1)2###37###40###41###0.01 %

NMCC-P1###4.34###+###+###+###W

NMCC-PT###5.11###+###+###+###W

NMCC-P3n###6.44###+###+###+###W

NMCCP7###6.87###+###+###+###W

NMCC4###6.12###+###+###+###W

NMCCP8###6.09###+###+###+###W

NMCCP-###6.81###+###+###+/-###W

NMCC-F###6.79###+###+###+###W

NMCC8###6.70###+###+###+###W

NMCC9###6.51###+###+###+###W

NMCC9###6.61###+###+###+/-###W

NMCC10###6.49###+###+###+/-###W

RESULTS AND DISCUSSION

Study samples from buffalo feces (BF) showed total Lactobacillus 5.2 +-.0.72 log CFU/g in early while 4.2 +-0.14 log CFU/g in middle and 4.1 +-0.14 log CFU/g in late lactating buffalo. As far as the Lactococcus species are concerned, 5.1+-0.72 CFU/g in early 4.5 +-0.14 and 4.1+-0.14 log CFU/g both in middle and late lactating respectively. In addition, total Enterococcus numbers in fecal samples were counted at levels between 5.1+-0.24 in early 5.3+-0.80 in middle and 5.0+-0.31 in late lactation stage. As far as the Bafidiobactrium are concerned, higher numbers (6.5+-0.31 CFU/g) were detected initially at early lactation (Figure 1). Nonetheless, Shakira et al. (2018) previously described the similar results of Lactobacillus counts (6.5 CFU/g) for cattle manure. In present study, thirty (30) different bacterial colonies were isolated from buffalo fecal samples.

After examination of these isolates through sample microscopy and electron microscopy which shown in fig 4 and 5 a total of twenty isolates, found as gram positive, catalase and oxidase negative (Table 1 and 8) were used for antimicrobial testing. L. monocytogenes, S. aureus, E.coli, B. cereus and P. Aeroginosa (ATCC) strains were used as indicator pathogens as shown in table 2 (Fijan, 2016). Six strains (NMCC-PI, NMCC-PT, NMCC-P3n, NMCC-P7, NMCC-P8 and NMCC-F) showed positive antibacterial activity (Table 3). These strains inhibited the growth of pathogenic bacteria particularly food borne pathogens including B. cereus, E. coli and L. monocytogenes. All six strains showed antagonistic effects towards tested pathogenic strains, however, the activity of antagonism was different among tested strains. NMCC-PI, NMCC-P7 and NMCC-P8 strains possessed strong activity against L. monocytogenes.

No significant activity of NMCC-PI was observed against P. aeruginosa and E.coli. Two isolated strains viz NMCC-P7 and NMCC-P8 had capacity to inhibit a number of pathogenic bacteria. Therefore, these two best strains (NMCC-P7 and NMCC-P8) could further be exploited as potential antimicrobial probiotic candidates against animal pathogens and be considered for their health implications. Antimicrobial activity of LAB towards pathogens may be due to production of the metabolites like, organic acids, hydrogen peroxide, bacteriocins (Lahtinen et al., 2009). Molecular identification of the presumptive probiotic bacterial strains is very crucial step to access and evaluate the safety of the microbes of an animal origin.

These isolates were molecular identified as Lactobacillus fermentum, Pediococcus acidilactici, Pediococcus pentosaceus, Enterococcus faecalis, Enterococcus ratti and Enterococcus lactis (Table 4). Acid tolerance is a basic criteria for screening probiotics species. All LAB had a wide range of pH tolerance form 1 to 4.

However, NMCC-PT, NMCC-P8 and NMCC-P7 showed significantly (Pa$?0.05) good acid tolerance (Table 5). Antibiotics acceptability is another important criteria of a probiotics strains beneficial for human and animal consumption (FAO/WHO, 2001). In the present study, none of strains were entirely susceptible to all tested antibiotics. Most of the strains showed resistance to most antibiotics (Table 6; Figure 2). The strain NMCC-P7 was moderately susceptible to Ampicilin. However, NMCC-PT, NMCC-P8 and NMCC-P7 strain showed maximum resistant to all antibiotics except Chloramphenicol and Erythromycin as shown in fig 2. The LAB strains showed resistance to penicillin G and Ampicillin in number of the studies Coppola et al., 2005; Ribeiro et al., 2014 and Papagianni and Anastasiadou 2009. In the present study, most of the strains were resistant to antibiotic Vancomycin, which inhibits cell wall synthesis.

This may be due to the presence of D-Ala-D-lactate in their peptidoglycan chains rather than the D-ala-D-ala dipeptide Meziane-Cherif et al., (2013) L. fermentum is capable of suppressing primarily gram-negative bacteria but to some extent it also suppresses Enterococci and S. aureus. P. acidilactici in addition to ordinary properties, like resistance to heat, acid and bile tolerance, has the ability to produce antimicrobial peptides with antimicrobial spectrum, which makes it an interesting potent probiotic (Papagianni and Anastasiadou 2009). The Pediococcus strains showed the maximum temperature tolerance (Ribeiro et al., 2014).

The capability of the bacterial strains to stand varying acidic, bile, pancreatic enzymes conditions and to adhere to intestinal epithelial cells has been considered as promising indicator for the survival of a bacterial strain. One of the main aspects of probiotic strains is the cholesterol assimilation. Microbial strains required lipid derivatives during their growth pattern. These fatty acids played a vital role in the synthesis and protection of eukaryotic cell membranes. In present study Cholesterol assimilation assay showed significant variation among microbial strains with better results in NMCC-P8 (Table 7). Present findings depicted that all six isolates showed good acid tolerance at pH 3; however, capacity of tolerance varied among the strains. All strains especially P. Acidilactici strains showed positive cellulolytic, proteolytic and amylolytic activity (Table 6; Figure 3).

P. acidilactici strains isolated from the buffalo GIT showed best probiotic results. Dairy industry has been facing a lot of problems worldwide, one of which is the use of unidentified microbial based feed products (Shakira et al., 2016; Lahtinen et al., 2009). Identification of the microbial based feed though latest molecular methods can generate unique information for the preparation of the animal probiotic. Present study was reported for the isolation and molecular identification of the animal based microbial flora. In the present study, 6 lactic acid bacterial strains were isolated from Nilli Ravi buffalo fecal samples. All strains showed good probiotic potential but overall results showed that the P. acidilactici can be used as probiotic for animal use.

Acknowledgements: This research was a part of an on-going project (Development of Buffalo Probiotic) funded by Punjab Agriculture Research Board (PARB-1002), Lahore, Pakistan

REFERENCES

Bergey's Manual of Determinative Bacteriology (7 th ed.). Robert, S. B., E. G. D. Murray and R. S. Nathan (1962). Baltimore, Md.: Williams and Wilkin. 1,094.

Coppola, R., M. Succi, P. Tremonte, A. Reale, G. Salzano and E. Sorrentino (2005). Antibiotic susceptibility of Lactobacillus rhamnosus strains isolated from Parmigiano Reggiano cheese. Le Lait. 85(3): 193-204.

Collado, M. C., E. Isolauri, S. Salminen and Y. Sanz (2009). The impact of probiotic on gut health. Curre. Drug. Metabo. 10 (1): 68-78.

Chen, W., H. Chen, Y. Xia, J. Zhao, F. Tian and H. Zhang (2008). Production, purification, and characterization of a potential thermostable galactosidase for milk lactose hydrolysis from Bacillus stearothermophilus. J. Dairy Sci. 91(5):1751-1758.

Del, R.B., B. Sgorbati, M. Miglioli and D. Palenzona (2000). Adhesion, autoaggregation and hydrophobicity of 13 strains of Bifidobacterium longum. Lett. Appl. Microbiol. 31:438-442.

Di-Gioia D., G. Mazzola, I. Nikodinoska, I. Aloisio, T. Langerholc, M. Rossi and J. Rovira (2016). Lactic acid bacteria as protective cultures in fermented pork meat to prevent Clostridium spp. growth. Intl. J. Food Microbiolo. 235: 53-59.

Di. Gioia, D., and B. Biavati (2018). Probiotics and Prebiotics in Animal Health and Food Safety: Conclusive Remarks and Future Perspectives. In Probio. Prebiot. Ani. Heal. Food Safe. 269-273: Springer, Cham.

FAO/WHO. (2002). WHO working group report on drafting guidelines for the evaluation of probiotics in food. London, Ontario, Canada, 30.

Fijan, S. (2016). Antimicrobial effect of probiotics against common pathogens, in Probiotics and Prebiotics in Human Nutrition and Health, In Tech.

Fuller, R. (1989). Probiotics in man and animals. J. applied bacteriolog. 66: 365-378.

Grochowska S., W. Nowak, R. Mikula and M. K. Potocka (2012). The effect of Saccharomyces cerevisiae on ruminal fermentation in sheep fed high-or low-NDF rations. J. of Ani and Feed Scien. 21(2): 276-284.

Gu. J., and K. Roberts (2019). Probiotics and Prebiotics. In Adult Short Bowel Syndrom. 67-80. Academic Press.

Halder, D., M. Mandal, S. Chatterjee and S. Mandal (2017). Indigenous probiotic Lactobacillus isolates presenting antibiotic like activity against human pathogenic bacteria. Biomedici. 5(2): 31.

Hassanzadazar, H., A. Ehsani, K. Mardani and J. Hesari (2012). Investigation of antibacterial, acid and bile tolerance properties of lactobacilli isolated from Koozeh cheese. In Veterin. Resear. Forum. 3 (3): 181. Faculty of Veterinary Medicine, Urmia University, Urmia, Iran.

Kvan, O. V., I. A. Gavrish, S. V. Lebedev, A. M. Korotkova, E. P. Miroshnikova, V. Serdaeva and N. Davydova (2018). Effect of probiotics on the basis of Bacillus subtilis and Bifidobacterium longum on the biochemical parameters of the animal organism. Environme. Scien. and Polluti. Resear. 25(3): 2175-2183.

Lahtinen, S. J., L. Tammela, J. Korpela, R. Parhiala, H. Ahokoski, H. Mykkanen and S. J. Salminen (2009). Probiotics modulate the Bifidobacterium microbiota of elderly nursing home residents. (1): 59-66.

Latorre J. D., X. Hernandez-Velasco, R. E. Wolfenden, J. L. Vicente, A. D. Wolfenden, A. Menconi and G. Tellez (2016). Evaluation and selection of Bacillus species based on enzyme production, antimicrobial activity, and biofilm synthesis as direct-fed microbial candidates for poultry. Front Veterin. Sci. (3): 95.

Lee Y. K., and S. Salminen (2009). Handbook of probiotics and prebiotics. John Wiley and Sons.

Markowiak, P., and K. Slizewska (2018). The role of probiotics, prebiotics and synbiotics in animal nutrition. Gut pathoge. 10(1): 21.

Meziane-Cherif D., P. J. Stogios, E. Evdokimova, A. Savchenko and P. Courvalin (2014). Structural basis for the evolution of vancomycin resistance D, D-peptidases. Proceedings of the National Acade. of Scienc. 111(16): 5872-5877.

Naeem, M., I. Ahmed, S. Ahmed, M.N. Riaz and S. Ghazanfar (2018). Screening of cattle gut associated Bacillus strains for their potential use as animal probiotic. Indian J. Anim. Res.1-6

Olatunde O. O., A. O. Obadina, A. M. Omemu, O. B. Oyewole, A. Olugbile and O. O. Olukomaiya (2018). Screening and molecular identification of potential probiotic lactic acid bacteria in effluents generated during ogi production. Annals of Microbiolo. 68(7): 433-443.

Papagianni, M., and S. Anastasiadou (2009). Encapsulation of Pediococcus acidilactici cells in corn and olive oil microcapsules emulsified by peptides and stabilized with xanthan in oil-in-water emulsions: Studies on cell viability under gastro-intestinal simulating conditions. Enzyme and Microbi. Technol. 45(6-7): 514-522.

Ribeiro, M. O., L. S. Vandenberghe, M. R. Spier, K. S. Paludo, C. R. Soccol and V. T. Soccol (2014). Evaluation of probiotic properties of pediococcus acidilactici b14 in association with lactobacillus acidophilus ATCC 4356 for application in a soy based aerated symbiotic dessert. Brazilian Archives of Biology and Technology. 57(5): 755-765.

Shakira, G., M. Qubtia, I. Ahmed, F. Hasan, M. I. Anjum and M. Imran (2018). Effect of indigenously isolated saccharomyces cerevisiae probiotics on milk production, nutrient digestibility, blood chemistry and fecal microbiota in lactating dairy cows. J. Anim. Plant. Sci. 28(2): 407-420.

Shakira. G, (2016). Study on the effect of dietary supplementation of Saccharomyces cerevisiae on performance of dairy cattle and heifers; PhD Thesis. Quaid-i-Azam University, Islamabad, Pakistan

Taylor, A. L., J. Dunstan and S.L. Prescott (2007). Probiotic supplementation for the first 6 months of life fails to reduce the risk of atopic dermatitis and increases the risk of allergen sensitization in high-risk children: a randomized controlled trial. J. of Alle. and Clini. Immun. 119 (1): 184-191.

Van, N., C. P. Oliszewski, S.N. Gonzalez and A.B. Perez Chaia (2007). Conjugated linoleic acid conversion by dairy bacteria cultured in MRS broth and buffalo milk. Lette. appli. microbe. 44 (5): 467-474.

Zeineldin, M., R. Barakat, A. Elolimy, A. Z. Salem, M. M. Elghandour and J. C. Monroy (2018). Synergetic action between the rumen microbiota and bovine health. Microb. Pathogene. 124: 106-115.
COPYRIGHT 2020 Knowledge Bylanes
No portion of this article can be reproduced without the express written permission from the copyright holder.
Copyright 2020 Gale, Cengage Learning. All rights reserved.

Article Details
Printer friendly Cite/link Email Feedback
Author:S. Khan, A-Ud-Din, G. M. Ali, S. I. Khan, I. Arif, M. N. Riaz and S. Ghazanfar
Publication:Journal of Animal and Plant Sciences
Date:Dec 31, 2020
Words:4098
Previous Article:INFLUENCE OF EXTRUDED CAMELINA SEED AND NATURAL COLOURANTS ADDITION IN LAYING HENS DIET ON EGGS YOLK COLOUR AND FATTY ACID COMPOSITION.
Next Article:THE POSSIBLE BENEFICAL EFFECT OF AMPELOPSIN ON INJURIES OF OVARIAN AND LUNG TISSUES GENERATEDBY OVARIAN TORSION/DETORSION.
Topics:

Terms of use | Privacy policy | Copyright © 2022 Farlex, Inc. | Feedback | For webmasters |