Enteric diseases are common on the first weeks of calves’ lives, negatively affecting ADG, and consequently increasing female heifers’ age at first conception (Svensson et al, 2003; Botteon et al., 2008). Additionally, diseases occurrence in neonatal period may cause future additional health problems, decreasing quality and quantity of milk production (Kozasa et al., 2005; Laureyns et al., 2009). Therefore, reduction in morbidity and duration of diarrhea can determine higher ADG and, consequently, improvements in animal health and production. In this study, we observed a shorter duration of diarrhea in the PHAGE group in the neonatal period and, consequently, a greater weight gain during the pre-weaned period.
Reduction in the diarrhea period and fewer calves with diarrhea up to 13 d of life in PHAGE may be associated with the control of bacterial proliferation provided by bacteriophage use. Co-infection is commonly seen in diarrheic calves (Cho and Yoon, 2014), and an intestinal dysbiosis and immune suppression may facilitate the proliferation of opportunistic bacteria increasing the duration and severity of the disease (Cho et al., 2013). Such fact was reported by Smith et al (1987) and Sulakvelidze et al. (2001), that confirmed the phage effect on reducing the magnitude of infectious processes. The less aggravated diarrheic episodes possibly justify serum protein and albumin levels increased significantly in the PHAGE group when compared to the CON after the 21st d of life. Also, calves supplemented with phage presented faster clinical recovery, less loss of protein, and possibly greater food intake, with better absorption of nutrients due to smaller extent of intestinal damage (Allison, 2017).
Loss of fluids and less nutrient absorption, associated with inflammatory process caused during diarrhea has a negative correlation with animals' weight gain (Barrington et al., 2002; Gifford et al., 2012). Thus, a shorter diarrhea duration may explain the higher ADG of calves that received milk replacer added with phages. It is noted that this difference is even greater considering only the neonatal period, which is the moment of the highest incidence of diarrhea (García et al., 2000).
In a study conducted with 122 dairy herds in Sweden, calves with neonatal diarrhea were more likely to develop respiratory infections up to six months of age (Hultgren et al., 2008). However, this association was not found in some herds in the USA and in Canada (Windeyer et al., 2014). In the present study, the use of phages was not able to reduce the incidence of respiratory disease, possibly because even reducing the duration of diarrhea, there was still a load of pathogens being eliminated in the environment.
Phage therapy probably acted on bacterial co-infection and secondary bacterial enteritis (Cho et al., 2013), however neonatal diarrhea may be caused by other factors or agents not controlled by phage use, such as nutrition, environment and virus (Cho and Yoon, 2014). Regarding causes of neonatal diarrhea, Santin et al. (2014) observed that 50.3% of fecal samples of calves between 5 days to 2 months of age were positive for Cryptosporidium spp., increasing to 66.7% when considering only animals up to 2 weeks of age. Thus, diarrhea prevalence was not reduced in this study probably due to primary causing factors or agents other than bacteria. However, phage therapy was able to decrease diarrhea duration by controlling opportunistic bacterial infections.
In vitro studies have shown satisfactory effects on the reduction of E. coli O: 157:H7 occurrence (Niu et al., 2009; Coffey et al., 2011; Carter et al., 2012), however, in vivo experiments show contradictory results. The phage therapy was effective in controlling E. coli intestinal population in cattle (Sheng et al., 2006; Niu et al., 2008) and sheep (Callaway, 2008). Nonetheless, other studies did not find the same effect (Bach et al., 2003; Raya et al., 2006; Rivas et al. 2010). Bacteria quantification was not possible in this study, but we believe that the reduction in the diarrheic episode duration in PHAGE was promoted by some decrease in the pathogenic bacteria burden.
Still, the success of the phage therapy may be associated with administration route. Ronzema et al. (2009) demonstrated that oral route is superior to intra-rectal. Sheng et al. (2006) observed that the rectal application (4 doses of 25 ml with 1x1010 CFU) was sufficient to control bacterial population, although in this study continuous administration of phages in lower concentrations in the water was maintained (1.8x106 CFU). Also, Anand et.al (2015) found that broad-spectrum phage was able kill 47.3% of E. coli isolates from diarrheic calves.
In this study, lytic bacteriophage added to milk replacer managed to control diarrhea, as observed by the decrease in the permanence of diarrheic feces during neonatal period. Additionally, greater weight gain in the first 80 d of life was obtained for calves supplemented with bacteriophage.