Calf diarrhoea during early life leads to huge economic losses in dairy farms (Cho and Yoon, 2014). New born calf requires critical care and attention but it has been neglected by many dairy farmers. Since the gastrointestinal tract of the calf is sterile, it becomes vulnerable to many enteropathogens e.g. Escherichia Coli, Salmonella spp. rotavirus, bovine corona virus Cryptosporidium parvum etc. (Abou El-Ella et al., 2013) often leading to increased mortality and morbidity (Islam et al., 2015). Antibiotic have been used with success but it decreases these beneficial bacteria (Ubeda et al., 2010) with increases the emergence of antibiotic resistance strains. Microbial colonization begins immediately after birth from the environment, fecal and vaginal flora during parturition which holds great importance in maturation of the gastrointestinal tract (Favier et al., 2002). Maintaining healthy microbial balance in calves leads to less immune disturbances as well as decrease health risk with optimal growth and production (Hill and Artis, 2009). Therefore, reduction of antibiotics with improved gut microbial balance can be introduced by feeding these good microbes during such colonization period (Malmuthuge et al., 2015). Synergistic effects was observed when probiotics and prebiotics are used simultaneously (Sekhon and Jairath, 2010). Therefore, the main objective of the experiment was to the study the effect of synbiotic on growth and health performance of the Jersey crossbred calves.

Materials and Methods                                                               

The study was carried out at ICAR, ERS-National Dairy Research institute, Kalyani, West Bengal. A total of twelve newborn Jersey crossbred calves were randomly allotted equally into two groups i.e. control and treatment group. The calves were separated from the dam immediately after birth and raised in groups up to 90 days of age. Colostrum was fed @1/10^th of the body weight within one hour after birth followed by a second dose after 10-12 hour up to 3 days of age.  Milk was bottle fed in two divided doses; morning (08.00 am) and evening (16.00 pm) hour up to 2 months of age @ 1/10^th of body weight and 1/20^th of the body weight on the 3^rd month. Ad libitum supply of concentrate, green fodder and clean drinking water was available at all times. Treatment group was fed 100ml synbiotic consisting Lactobacillus rhamnosus NCDC 298 (10⁹ CFU/ml) and Fructo-oligosaccharide (10%) mixed with colostrum and milk up to 42 days of age (Jenny et al., 1991 and Cruywagen et al., 1996).

Dry matter intake (DMI), body weight and body measurement were recorded at weekly interval. Dry matter from milk, concentrate and green fodder were analyzed as per AOAC (2005) on weekly basis. Faecal score was determined on the basis of faecal consistency which was observed daily by using a numerical score of 1 to 4 (1= normal consistency, 2= slightly liquid consistency, 3= moderately liquid consistency, 4= primarily liquid consistency) as proposed by Morrison et al. (2010). Faecal samples were collected at weekly intervals and stored at -20^oC for Lactobacillus load count and 10% formalin for quantitatively  examination of faecal parasitic egg (Oocyst per gram, OPG) count using a Modified McMaster slide (Soulsby, 1982). Lactobacillus sp. load count was determined by diluting one gram of faecal sample in 9ml of 0.9% normal saline and tenfold dilutions were made up to 10^-9 dilution.1ml of mixed dilution from each tube were transferred in respective petri dishes. For culture of Lactobacillus, MRS agar (Himedia®) was used by pour plate technique and incubated at 37^oc for 24- 48 hour. Colonies having 30-300 colony count were counted and number of colony forming units (CFU) was expressed as log colony-forming units per gram (log CFU/g) of faeces.

Statistical Methods

All the data were analyzed using SPSS; Analysis of variance (ANOVA) was carried out to analyze the data. The analysis of oocyst per gram (OPG) data was carried out by least-squares analysis of variance (Harvey, 1990). The faecal oocyst counts were log-transformed in order to stabilize variance. The results were back-transformed by taking antilogarithm of least-squares means (LSM) and presented as Geometric means of faecal oocyst count (GFOC).

Results and Discussion

The results obtain for dry matter intake, body weight and body measurements are given in Table 1.

Table 1: Effect of synbiotic on DMI, body measurements and body weight of Jersey crossbred calves during 90 days of experiment period

^xy differences in superscript in row indicate significance at P<0.05

The results indicated that dry matter intake/100 kg body weight did not differ between the groups. This may be due to similar feeding and management conditions of the experimental calves though it was expected to be higher in the treatment group. Roodposhti and Dabiri (2012) studied effect of feeding multi-strain probiotics with prebiotic (Tipax) in claves and found no significant effects in weekly dry matter intake. Various workers, after feeding probiotics such as L. acidophilus 27SC (Abu-Tarboush et al., 1996), Lactobacillus acidophilus (Cruywagen et al., 1996), probiotic containing Lactobacillus spp. (Agazzi et al., 2014), mixture of Lactobacillus strains (Bayatkouhsar et al., 2013), L. plantarum and Bacillus subtilis (Zhang et al., 2016) in Holstein calves found no changes in the DMI. On the contrary, Gupta et al. (2016) found significant higher dry matter intake in the calves supplemented with Lactobacillus acidophilus and Lactobacillus plantarum. The result obtain from body measurement (cm) included body length, heart girth and wither height had no significant differences between the groups although overall gain in the respective measurements was numerically on higher side for the synbiotic fed group. The increasing trend was also reported by Nageshwar et al. (2016) without any significant differences between the groups of calves fed with or without probiotic. Higginbotham et al. (1998), Heinrichs et al. (2009), Riddell et al. (2010) observed similar results in the calves fed with probiotics.

The initial body weight was similar in both the groups whereas final body weight (kg) was significantly (P<0.01) higher in the treatment group (52.50±2.25). Average daily gain (ADG; g) was also significantly higher (P<0.05) in the synbiotic fed group (323.14 ± 0.01) when compared to control group (262.03± 0.03). The results are in agreement with the results of Roodposhti and Dabiri (2012) who reported significantly higher average daily gain (g) in the calves fed synbiotic. Increase in body weight due to synbiotic supplementation in calves was also reported by Hasunuma et al. (2011) and Dar et al. (2017). Besides synbiotic use, probiotics such as, L. plantarum and L. acidophilus (Gupta et al., 2016), L. sporogenes and S. cerevisiae (Nageshwar et al., 2016) etc. have reported significant increase in body weight gain and average daily gain in the calves. However, Kawakami et al. (2010) found no significant effects of lactic acid bacteria in the body weight gain of calves. The result obtain from Lactobacillus load count, faecal score, disease incidences and OPG count are given in Table 2.

Table 2: Effect of synbiotic feeding on recovery of Lactobacillus in feces, disease incidences, fecal score and parasitic oocyst count

^(abc) Means with different superscripts in a row differ significantly at (P<0.01); ^xyMeans with different superscripts in a row differ significantly irrespective of week (P<0.05)

The result indicated that recovery of Lactobacillus was significantly higher (P<0.01) throughout the study period in the calves fed synbiotic as compare to control group this may be due to colonization and development of Lactobacillus sp. in the intestine with intestinal microbial balance. The results are in agreement with the findings of Quezada-Mendoza et al. (2011) who also found significant higher Lactobacillus count in calves fed Lactobacillus and Propionibacterium spp. Similarly, Lee et al. (2012) reported higher faecal Lactobacillus load count after offering Latobacillus plantarum and Bacillus subtilis. Oli et al. (1998) reported recovery of Lactobacillus in pigs fed with Fructo-oligosaccharide. However, Hasunuma et al. (2011) reported no effect of synbiotic feeding on fecal Lactobacillus sp. count.

During the study period of 90 days, no disease incidences were observed other than diarrhoea and fever in both the groups. The number of calves suffered due to diarrhea (incidence days) was n=4/6 (9 days) in treatment and n=5/6 (8 days) in control groups respectively. Timmerman et al. (2005) and Mokhber-Dezfouli et al. (2007) also who also reported significant decrease of calf diarrhea when treated with lactic acid bacteria. Besides diarrhea, the experimental calves also suffered from fever though the numbers were same n=4, the sick days was 4 in control and 2 in the treatment group respectively.  Likewise, Maldonado et al. (2017) reported that the calves treated with lactic acid fermented milk did not suffer from fever. Better faecal score was observed in the treatment group. It was significantly (P<0.05) higher in control group (1.07±0.02) than the calves in the treatment group (1.02±0.01). This might be due to Lactobacillus strains which interfere with the adherence and development of pathogenic microorganisms by maintaining tight junction in the intestine (Berg, 1996). The findings corroborated with Hasunuma et al. (2011) who observed better faecal score in the calves fed synbiotic. Lee et al. (2012) also reported significantly better faecal score when claves were fed Lactobacillus plantarum and Bacillus subtilis. Similarly, Bayatkouhsar et al. (2013) and Agazzi et al. (2014) observed reduced faecal score in calves treated with Lactobacillus strains compared to control group.

The overall mean log transformed OPG count (GFOC) during 90 days experimental trail was 5.56±0.16 (106) and 5.33±0.16 (106) in control and treatment group respectively and no significant differences have been observed between the groups although it was numerically lesser in the synbiotic fed group. Several workers have found non-significant effect on OPG counts after feeding lactic acid producing bacteria (Harp et al., 1996), mixture of probiotics (Higginbotham et al., 1998), mannan oligosaccharide (Terre et al., 2007) and Lactobacillus sp. (Frizzo et al., 2010).

Conclusion

Synbiotic supplementation enhanced body weight gain and ADG in Jersey crossbred calves. However, there was no change in dry matter intake and body measurements. Synbiotic feeding also reduced the number of sick calves with less number of days suffered and better faecal score. Though non-significant, the synbiotic fed calves also had lower OPG count. It may be concluded that supplementation of synbiotic had beneficial effects on the growth and health of the calves.

Acknowledgements

Authors are acknowledged to Director of National Dairy Research Institute, Karnal and all staff members of Eastern regional station, National Dairy Research Institute, Kalyani for providing assistance and necessary facilities to conduct research.

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