The Effect of a Direct Fed Microbial on Liveweight and Milk Production in Dairy Cattle.

This longitudinal study aimed to quantify the effects of dietary supplementation of a direct-fed microbial (DFM) consisting of three lactobacilli isolates on milk yield, milk fat and protein yields, somatic cell count (SCC), and liveweight in a single dairy herd in Australia. A total of 150 dairy cows were randomly selected based on parity and days in milk and divided into two groups: control (n = 75) and DFM treatment (n = 75). Throughout the study, the two groups of cows were housed separately in a dry lot yard, and each group had their own feeding area. For the DFM treatment group, selected cows in mid-lactation were supplemented with 10 mL/cow/day of the DFM via top dressing of the feed for the remainder of the lactation and through the dry period, extending into subsequent lactation. The control group had no supplementation. The milk yield and liveweight were recorded daily. Milk samples were collected every two months for milk component analysis (fat, protein, and somatic cell count [SCC]). The DFM-treated cows gained more liveweight across the study (19.40 kg, 95% CI 0.44 kg; 38.30 kg, p = 0.05) compared to the control cows. In the second production year, the DFM-treated cows mobilized more liveweight (-6.06 kg, 95% CI -10.49 kg; -1.61 kg, p = 0.01) and produced more milk (0.39 L/d 95% CI 0.10; 0.89, p = 0.05). Over a full lactation, DFM cows yielded at least 258 L (95% CI 252 L; 265 L) more milk than controls. No significant differences were found in fat and protein yield or SCC. This study suggests that consistent and ongoing supplementation with a Lacticaseibacillus- and Lentilactobacillus-based DFM could have a positive effect on milk production, but further research is needed to understand the underlying mechanism.

Effect of a Lactobacilli-Based Direct-Fed Microbial Product on Gut Microbiota and Gastrointestinal Morphological Changes.

The calf's gastrointestinal tract (GIT) microbiome undergoes rapid shifts during early post-natal life, which can directly affect calf performance. The objectives of this study were to characterise and compare differences in the establishment and succession of GIT microbiota, GIT morphological changes, and the growth of dairy calves from birth until weaned. Forty-four newborn Holstein-Friesian calves were randomly selected and assigned to Treatment (TRT) and Control (CON) groups. The TRT group calves received a once-daily dose of a direct-fed microbial (DFM) liquid product containing Lacticaseibacillus paracasei, Lentilactobacillus buchneri, and Lacticaseibacillus casei, all formerly known as Lactobacillus. Fresh faecal samples were manually taken from the rectum of all calves, and gross necropsy was performed on the forestomachs and gastrointestinal tracts. Bacterial DNA was extracted from frozen faecal samples for 16S rRNA gene amplicon sequencing. Calves in the TRT group had greater live weights (p = 0.02) at weaning compared with calves in the CON group (mean = 69.18 kg, SD = 13.37 kg). The average daily live weight gain (ADG) and total feed intake were similar between the two groups. Calves in the TRT group had greater duodenum, abomasum, and reticulum weights (p = 0.05). Rumen and intestinal development (p < 0.05) and faecal microbial diversity (p < 0.05) were more pronounced in the TRT group. The relative abundances of eight genera differed (p < 0.001) between the groups. Supplementing calves with the LAB-based DFM increased live weight at weaning and had a more pronounced effect on the development of rumen and the gastrointestinal tract and on microbiota diversity and evenness. Future work is needed to better understand the potential association of LAB-DFM products on gut mucosa-associated microbiota.

The Effect of Direct-Fed Lactobacillus Species on Milk Production and Methane Emissions of Dairy Cows.

Using direct-fed microbials to mitigate enteric methane emissions could be sustainable and acceptable to both consumers and producers. Forty lactating, multiparous, Holstein-Friesian cows were randomly allocated one of two treatments: (1) a base of ad libitum vetch (Vicia sativa) hay and 7.0 kg DM/d of a grain mix, or (2) the basal diet plus 10 mL of MYLO® (Terragen Biotech Pty Ltd., Coolum Beach, Queensland, Australia) delivering 4.17 × 108 cfu of Lactobacillus per mL. Neither feed intake (25.4 kg/d vs. 24.8 kg/d) nor milk yield (29.9 vs. 30.3 kg/d) were affected by treatment. Feed conversion efficiency was not affected by treatment when expressed on an energy-corrected milk basis (1.15 vs. 1.18 kg/kg DMI). Neither methane yield (31.6 vs. 31.1 g/kg DMI) nor methane intensity (27.1 vs. 25.2 g/kg energy corrected milk) were affected by treatments. While these results are contrary to our expectations and not significant, all were numerically in a favorable direction. Given there are reports that diet and dose rate may impact the size of any effect, we recommend a dose-response study be undertaken using a basal diet that is commonly used in pasture-based dairy systems.

Systematic review and meta-analysis of probiotic use on inflammatory biomarkers and disease prevention in cattle.

The aim of this study was to appraise the available evidence on the effectiveness of probiotic treatment on mature cattle immunity, inflammation, and disease prevention. A systematic review with meta-analysis was conducted to analyse studies that were eligible to answer the following research question: "in cattle of at least 6-months of age, is the use of probiotics associated with immunomodulatory and inflammatory responses, and clinical disease outcomes?" Our literature search yielded 25 studies that fit the inclusion criteria. From these studies, only 19 were suitable for inclusion in the meta-analysis due to data limitations and differences in study population characteristics. Included studies were assessed for bias using a risk assessment tool adapted from the Cochrane Collaboration's tool for assessing risk of bias in randomised trials. GRADE guidelines were used to assess the quality of the body of evidence at the outcome level. The meta-analysis was performed using Review Manager and R. The overall quality of evidence at the outcome level was assessed as being very low. On average, the treatment effect on immunoglobulin G (IgG), serum amyloid A (SAA), haptoglobin (Hp) and ß-hydroxybutyrate (BoHB) for cows receiving probiotics did not differ from control cows. Exposure to probiotics was not associated with reduced risk of reproductive disorders (pooled RR = 1.02 95 % CI = 0.81-1.27, P = 0.88). There is insufficient evidence to support any significant positive effects of probiotics on cattle immunity and disease prevention. This lack of consistent evidence could be due to dissimilarities in the design of the included studies such as differences in dosage, dose schedule, diet composition and/or physiological state of the host at the time of treatment.

Sequence characterisation and novel insights into bovine mastitis-associated Streptococcus uberis in dairy herds.

Streptococcus uberis is one of the most frequent mastitis-causing pathogens isolated from dairy cows. Further understanding of S. uberis genetics may help elucidate the disease pathogenesis. We compared the genomes of S. uberis isolates cultured from dairy cows located in distinctly different geographic regions of Australia. All isolates had novel multi locus sequence types (MLST) indicating a highly diverse population of S. uberis. Global clonal complexes (GCC) were more conserved. GCC ST86 and GCC ST143 represented 30% of the total isolates (n = 27) and were clustered within different geographic regions. Core genome phylogeny revealed low phylogenetic clustering by region, isolation source, and MLST. Identification of putative sortase (srtA) substrates and generation of a custom putative virulence factor database revealed genes which may explain the affinity of S. uberis for mammary tissue, evasion of antimicrobial efforts and disease pathogenesis. Of 27 isolates, four contained antibiotic resistance genes including an antimicrobial resistance cluster containing mel/mef(A), mrsE, vatD, lnuD, and transposon-mediated lnuC was also identified. These are novel genes for S. uberis, which suggests interspecies lateral gene transfer. The presence of resistance genes across the two geographic regions tested within one country supports the need for a careful, tailored, implementation and monitoring of antimicrobial stewardship.

Bioactive Streptomycetes from Isolation to Applications: A Tasmanian Potato Farm Example.

The genus Streptomyces constitutes approximately 50% of all soil actinomycetes, playing a significant role in the soil microbial community through vital functions including nutrient cycling, production of bioactive metabolites, disease-suppression and plant growth promotion. Streptomyces produce many bioactive compounds and are prime targets for industrial and biotechnological applications. In addition to their agrobiological roles, some Streptomyces spp. can, however, be phytopathogenic, examples include, common scab of potato that causes economic losses worldwide. Currently used chemical control measures can have detrimental effect to environmental and human health as a result alternative methods to chemical disease control are being investigated. One alternative is the use of streptomycete specific phages to remove this pathogenic bacterium before it can cause the disease on potatoes. However, due to co-existence of non-common scab-causing species belonging to the genus Streptomyces, phage treatment is likely to affect a wide range of non-target streptomycete species including the beneficial ones in the soil. Therefore, before such treatment starts the host range of the phages within the targeted family of bacteria should be determined. In a study conducted using soil samples from a Tasmanian potato farm, streptomycetes were isolated and tested against streptomycete-specific phages. Their antifungal activity was also determined using multiple assays against selected phytopathogens. The four strongest antifungal activity-displaying isolates were further tested for their persistent antifungal activity using wheat and Fusarium solani in a pot trial. A second pot trial was also conducted to evaluate whether the beneficial streptomycetes were affected by streptophage treatment and whether their removal via the phage battery would cause opportunistic fungal infections to plants in soil. The streptomycetes prevented the reduction in wheat shoot weight caused by F. solani indicating their disease suppressive effect. However, when phages were added into the pots, the growth of wheat was detrimentally impacted. This finding might suggest that the reduced presence of antifungal streptomycetes via phage-induced lysis might encourage opportunistic fungal infections in plants.

Survey and Sequence Characterization of Bovine Mastitis-Associated Escherichia coli in Dairy Herds.

Escherichia coli is frequently associated with mastitis in cattle. "Pathogenic" and "commensal" isolates appear to be genetically similar. With a few exceptions, no notable genotypic differences have been found between commensal and mastitis-associated E. coli. In this study, 24 E. coli strains were isolated from dairy cows with clinical mastitis in three geographic regions of Australia (North Queensland, South Queensland, and Victoria), sequenced, then genomically surveyed. There was no observed relationship between sequence type (ST) and region (p = 0.51). The most common Multi Locus Sequence Type was ST10 (38%), then ST4429 (13%). Pangenomic analysis revealed a soft-core genome of 3,463 genes, including genes associated with antibiotic resistance, chemotaxis, motility, adhesion, biofilm formation, and pili. A total of 36 different plasmids were identified and generally found to have local distributions (p = 0.02). Only 2 plasmids contained antibiotic resistance genes, a p1303_5-like plasmid encoding multidrug-resistance (trimethoprim, quaternary ammonium, beta-lactam, streptomycin, sulfonamide, and kanamycin) from two North Queensland isolates on the same farm, while three Victorian isolates from the same farm contained a pCFSAN004177P_01-like plasmid encoding tetracycline-resistance. This pattern is consistent with a local spread of antibiotic resistance through plasmids of bovine mastitis cases. Notably, co-occurrence of plasmids containing virulence factors/antibiotic resistance with putative mobilization was rare, though the multidrug resistant p1303_5-like plasmid was predicted to be conjugative and is of some concern. This survey has provided greater understanding of antibiotic resistance within E. coli-associated bovine mastitis which will allow greater prediction and improved decision making in disease management.

Description of Behavioral Patterns Displayed by a Recently Weaned Cohort of Healthy Dairy Calves.

Animals display movement patterns that can be used as health indicators. The movement of dairy cattle can be characterized into three distinct cluster types. These are cluster type 1 (resting), cluster type 2 (traveling), and cluster type 3 (searching). This study aimed to analyze the movement patterns of healthy calves and assess the relationship between the variables that constitute the three cluster types. Eleven Holstein calves were fitted with GPS data loggers, which recorded their movement over a two week period during spring. The GPS data loggers captured longitude and latitude coordinates, distance, time and speed. It was found that the calves were most active during the afternoon and at night. Slight inconsistencies from previous studies were found in the cluster movements. Cluster type 2 (traveling) reported the fastest rate of movement, whereas cluster type 1 (resting) reported the slowest. These diverse movement patterns could be used to enhance the assessment of dairy animal health and welfare on farms.

Efficacy of a Lactobacillus-Based Teat Spray on Udder Health in Lactating Dairy Cows.

Teat disinfection is a common pre- and post-milking mastitis prevention practice that is part of a mastitis control program in dairy herds. Commercially available teat disinfectants are generally chemical-based products. The use of these products has occasionally raised concerns about the risk of chemical residues in milk. An alternative treatment or prevention strategy based on probiotics has the potential to circumvent this risk. Two treatments were compared in a cross-over clinical trial in a single herd: a lactobacillus-based, post-milking teat spray (LACT), and a commercial iodine-based post-milking teat disinfectant product as (positive control, PC). The effect of the two treatments on cow somatic cell counts was quantified using a multivariate mixed-effects linear regression model with cow fitted as a random effect. The odds of teat end scores increasing from a low to a high score tended to be lower (OR = 0.74, 95% CI 0.54-1.01, P = 0.06) for cows receiving LACT treatment. On average, there was also a tendency for a lower somatic cell counts in the LACT treated cows (antilog of coefficient = 0.91, 95% CI 0.80-1.03, P = 0.13) compared with the PC treated cows. The application of the lactobacillus-based product to teats could reduce the rate of teat end scores progression from low to higher scores, and potentially improve teat end sphincter functions and udder health. Further, larger scale validation work is required to support the findings of the current study.

Systematic review of an intervention: the use of probiotics to improve health and productivity of calves.

The aims of this study were to undertake a systematic review and meta-analysis of the types of probiotic formulations that are commercially available and to critically appraise the available evidence for the effectiveness of probiotics in improving the health and productivity of calves. Relevant papers were identified to answer the question: 'In calves aged between birth to one year, is the use of probiotics associated with changes in haematological or biochemical parameters, faecal bacteria counts, average daily live weight gain, dry matter intake, or feed conversion ratio?' The search of the literature yielded 67 studies that fit the primary screening criteria. Included studies were assessed for bias and confounding using a predefined risk assessment tool adapted from the Cochrane Collaboration's tool for assessing risk of bias in randomised trials and GRADE guidelines. Meta-analysis was performed using Review Manager and R. Random sequence generation was low in more than 59 % of studies. Risk of allocation concealment and performance bias were largely unclear in over 68 % of studies. Calves fed probiotics had increased average daily live weight gains (ADG) from birth to weaning (mean difference [MD] = 83.14 g/d 95 % CI = 58.36-107.91, P < 0.001) compared with calves on a control diet. Calf age reduced the level of heterogeneity of the effect of probiotics on ADG for calves between one to three weeks of age (τ2 = 73.15; I2 = 4%; P = 0.40) but not for calves older than three weeks of age (τ2 = 2892.91; I2 = 73 %; P < 0.001). Feed conversion ratio (FCR) was lower for calves on probiotics (MD = -0.13 kg of dry matter intake (DMI) to kg of live weight (LW) gain, 95 % CI = -0.17 to -0.09, P < 0.001), and the heterogeneity of effect was large in younger aged calves (τ2 = 0.05; I2 = 78 %; P = 0.03). The risk of bias regarding the methodology in the included studies was high. The quality of evidence for each outcome was categorised as moderate. There is sufficient data to support the effectiveness of probiotic use in some applications such as for the improvement of performance and productivity parameters of calves. However, the evidence is weak for other potential probiotic uses in calves such as improved health and reduced risk of disease. Therefore, the existing data are inconclusive and do not support the use of probiotics as an alternative to antimicrobials to improve calf health and productivity.

Assessment of the Detrimental Impact of Polyvalent Streptophages Intended to be Used as Biological Control Agents on Beneficial Soil Streptoflora.

Streptophages are currently being investigated to control potato common scab, however, since a majority of streptophages are reported to be polyvalent, their potential to infect beneficial soil streptomycetes during the application process may have unintended consequences. To test this hypothesis, two phytopathogenic fungi, namely Fusarium solani and Rhizoctonia solani, were tested for their detrimental effect on the test crop wheat (Triticum aestivum cv. Gutha). F. solani caused a significant root weight reduction (34%) in the wheat plant and therefore was tested further in the pot trials with actinomycetes present. Sixty-seven streptomycete isolates from a Tasmanian potato farm were screened for their antifungal abilities against the two phytopathogenic fungi. Four actinomycetes found to be strongly antifungal were then tested for their disease-protective abilities against F. solani in pot trials again using wheat. Addition of the streptomycetes into the container media protected the plants against F. solani, indicating that streptomycetes have a disease-suppressive effect. A further pot trial was conducted to evaluate whether these beneficial streptomycete species would be affected by streptophage treatment and subsequently result in an increased risk of fungal infections. When streptophages were added to the pots, the shoot and root growth of wheat declined by 23.6% and 8.0%, respectively, in the pots with the pathogenic fungus compared to the control pots. These differences might suggest that removal of antifungal streptomycetes by polyvalent phages from plant rhizosphere when biocontrol of plant pathogenic streptomycetes (e.g. Streptomyces scabiei) is targeted might encourage secondary fungal infections in the farm environment. The presented data provide preliminary evidence that streptophage treatment of pathogenic streptomycetes may lead to an aggravated disease risk by soil-borne fungal pathogens when naturally present antagonists are removed. As a result, extensive farm site trials are required to determine the long-term detrimental impact of polyvalent streptophage treatments on beneficial soil streptoflora.