Molecular characterization of antimicrobial resistance genes on farms and in commercial milk with emphasis on the effect of currently practiced heat treatments on viable but nonculturable formation.

Despite the considerable advances that have been made to improve dairy food safety, there is rising concern that pasteurization is not sufficient for the destruction of plasmid-mediated antimicrobial resistance (AMR) genes of resistant bacteria and could stimulate bacteria to enter into a viable but nonculturable (VBNC) state. In the current study, we surveyed the prevalence of 1 genomic and 9 plasmid-mediated AMR genes in 100 samples (bulk tank milk and milk filter socks) at the farm level and 152 commercial milk samples (pasteurized and UHT milks) and assessed the VBNC state in dairy bacteria. Results revealed that sul2 was the most prevalent plasmid-mediated gene in milk filter socks (96%), bulk tank milk (48%), pasteurized milk (68%), and UHT (43%) milk; in contrast, mecA was not detected in any sample. Additionally, commercial pasteurization (as currently practiced) failed to decrease the prevalence of the blaTEM-B1 (43%), tetK (30%), and tetA (55%) plasmid-mediated AMR genes; thus, commercial pasteurization may be one of the factors creating the VBNC state in some dairy bacteria. Continued research is necessary to identify bacterial species entering the VBNC state after pasteurization, to assess their potential hazard level and shed more light on the expression and possibility of horizontal gene transfer of those plasmid-mediated AMR genes.

A large-scale study of indicators of sub-clinical mastitis in dairy cattle by attribute weighting analysis of milk composition features: highlighting the predictive power of lactose and electrical conductivity.

Sub-clinical mastitis (SCM) affects milk composition. In this study, we hypothesise that large-scale mining of milk composition features by pattern recognition models can identify the best predictors of SCM within the milk composition features. To this end, using data mining algorithms, we conducted a large-scale and longitudinal study to evaluate the ability of various milk production parameters as indicators of SCM. SCM is the most prevalent disease of dairy cattle, causing substantial economic loss for the dairy industry. Developing new techniques to diagnose SCM in its early stages improves herd health and is of great importance. Test-day Somatic Cell Count (SCC) is the most common indicator of SCM and the primary mastitis surveillance approach worldwide. However, test-day SCC fluctuates widely between days, causing major concerns for its reliability. Consequently, there would be great benefit to identifying additional efficient indicators from large-scale and longitudinal studies. With this intent, data was collected at every milking (twice per day) for a period of 2 months from a single farm using in-line electronic equipment (346 248 records in total). The following data were analysed: milk volume, protein concentration, lactose concentration, electrical conductivity (EC), milking time and peak flow. Three SCC cut-offs were used to estimate the prevalence of SCM: Australian ≥ 250 000 cells/ml, European ≥200 000 cells/ml and New Zealand ≥ 150 000 cells/ml. At first, 10 different Attribute Weighting Algorithms (AWM) were applied to the data. In the absence of SCC, lactose concentration featured as the most important variable, followed by EC. For the first time, using attribute weighted modelling, we showed that the concentration of lactose in milk can be used as a strong indicator of SCM. The development of machine-learning expert systems using two or more milk variables (such as lactose concentration and EC) may produce a predictive pattern for early SCM detection.

Repurposing Ionophores as novel antimicrobial agents for the treatment of bovine mastitis caused by Gram-positive pathogens.

Increasing reports of multidrug-resistant bacterial infections in animals has created a need for novel antimicrobial agents that do not promote cross-resistance to critically important antimicrobial classes used in human medicine. In response to the recent emergence of antimicrobial resistance in several bovine mastitis pathogens, in vitro antimicrobial susceptibility was determined for four polyether ionophores (lasalocid, monensin, narasin and salinomycin) against Staphylococcus spp. and Streptococcus spp. isolated from clinical cases. In addition, erythrocyte haemolysis and WST-1 cell proliferation assays were used to assess in vitro mammalian cell cytotoxicity and biofilm susceptibility testing was performed using the minimum biofilm eradication concentration (MBEC™) biofilm assay. Lasalocid, monensin, narasin and salinomycin exhibited bacteriostatic antimicrobial activity against all pathogens tested, including methicillin-resistant staphylococci, with MIC90 values <16 µg/ml. Narasin and monensin displayed the least toxicity against mammalian cell lines and all compounds significantly reduced viable cell numbers in a Staphylococcus aureus biofilm. Based on in vitro characterization, all four ionophores offer potentially novel treatments against bovine mastitis but in vivo studies will be essential to determine whether acceptable safety and efficacy is present following intramammary administration.

Development of an improved Streptococcus uberis experimental mastitis challenge model using different doses and strains in lactating dairy cows.

Developing a reliable mastitis challenge infection model is required to test new intramammary antimicrobial preparations, other novel bovine mastitis treatments, and study mastitis pathogenesis. Three treatment groups of Holstein Friesian cows in active lactation were administered two doses (10(4) and 10(6) cfu/quarter) on a single occasion with one of the three Streptococcus uberis strains (BFR6019, MFF1283 and SA002) suspended in 5 ml of sterile PBS, administered via intramammary inoculation immediately after milking. All quarters that were challenged with S. uberis strains MLF1283 and BFR6019 showed clinical signs of mastitis on day 1 and 2 after the challenge. Strain SA002 had a lower rate of inducing clinical mastitis which was detected later than day 3 after the challenge. We successfully developed a rapid and reliable model for inducing experimental S. uberis mastitis with 100% success rate in cows in active lactation. On the basis of the correlation results between strains, RAPD fingerprinting results, clinical findings, and a 100% success rate of mastitis induction for low and high doses S. uberis strains MLF1283 and BFR6019, strain virulence seems to be a more important effect than challenge dose in induction of clinical mastitis following experimental challenge.

Effect of Feeding a Calcium Chloride Supplement on Sow Stillbirth Rate.

The present study was undertaken to determine the effect of daily calcium chloride (CaCl2) supplementation from day of entry into the farrowing house until day of farrowing (6.4 ± 0.3 d) on stillbirth rates. Landrace × Large White sows (parities 4 to 6; n = 53) were offered 40 g/d CaCl2 (n = 28) or served as controls (n = 25). The morning before their estimated farrowing date, a blood sample was obtained from 25 sows for calcium measurement and a urine sample from 22 sows for pH measurement. The feeding of CaCl2 decreased urinary pH compared to the control group (p < 0.001), indicative of an induced metabolic acidosis, but there was no effect of feeding CaCl2 on serum calcium concentrations or the incidence of stillbirths. Nonetheless, regardless of treatment, sows with higher serum calcium concentrations (>2.5 vs. <2.5 mmol) or lower urine pH (<7.0 vs. >7.0) had fewer stillborn piglets (p < 0.001 for both). While showing that low serum calcium levels will increase stillbirth rates, our data indicate that the administration of 40 g/d CaCl2 for 6 d prior to farrowing was not sufficient to increase serum calcium or decrease stillbirth incidence.

Preweaning Purified Fucoidan Drench: Effects on Growth, Immune Response, and Intestinal Morphology in Weaned Piglets.

Weaning stress imposes considerable physiological challenges on piglets, often manifesting in intestinal disturbances, such as inflammation and compromised barrier function, ultimately affecting growth and health outcomes. While conventional interventions, including antimicrobials, have effectively mitigated these sequelae, concerns surrounding antimicrobial resistance necessitate the exploration of alternatives. Fucoidan, derived from brown seaweed, offers promise due to its antioxidant and anti-inflammatory effects. Previous research has been limited to the in-feed supplementation of partially purified fucoidan extracted from brown seaweed. The focus of the present study is assessing the effect of a preweaning drench with highly purified (85%) fucoidan on piglet growth, immune response, and intestinal morphology post-weaning. Forty-eight male piglets at 17 ± 3 days of age (5.67 ± 0.16 kg) were assigned to a saline (control), fucoidan, or antimicrobial group, receiving treatment as a single 18 mL oral drench three days before weaning. Monitoring for seven days post-weaning included body weight measurements, blood sample collection for the inflammatory protein assay, and small intestine morphological analysis. The findings revealed that the preweaning fucoidan drench did not elicit adverse effects on piglets. However, neither fucoidan nor antimicrobial drenches significantly enhanced growth parameters, immune markers, or intestinal morphology compared to that of the control-treated piglets (p > 0.05). The lack of response may be attributed to the high health status of the experimental cohort and the limitation of a single dosage. Future research should consider a more challenging production setting to evaluate the viability and optimal application of fucoidan as an antimicrobial alternative in the pig industry.

Application of Exogenous GnRH in Food Animal Production.

Over several decades, exogenous GnRH and agonists have been employed for controlling reproductive cascades in animals, and treating some reproductive morbidities. The administration of GnRH is used in animals to counter ovarian dysfunction, induce ovulation, and to increase conception and pregnancy rates. GnRH and its agonists are used in the treatment of cystic ovarian degeneration and repeat breeder syndrome. The development of protocols for GnRH administration by intramuscular injection, intramuscular or subcutaneous implants, and intravaginal deposition has empowered their clinical use worldwide. Currently, exogenous GnRH products are a central part of several pre- and post-breeding programs for the enhancement of fertility, including the control of estrous cycles and timing of ovulation, development of fixed-time artificial insemination protocols, improved embryo survival, and the treatment of reproductive morbidity. The aim of the present review is to summarize the application of exogenous GnRH agonists in food animal production.

Phylogeny, Virulence, and Antimicrobial Resistance Gene Profiles of Enterococcus faecium Isolated from Australian Feedlot Cattle and Their Significance to Public and Environmental Health.

The extent of similarity between E. faecium strains found in healthy feedlot beef cattle and those causing extraintestinal infections in humans is not yet fully understood. This study used whole-genome sequencing to analyse the antimicrobial resistance profile of E. faecium isolated from beef cattle (n = 59) at a single feedlot and compared them to previously reported Australian isolates obtained from pig (n = 60) and meat chicken caecal samples (n = 8), as well as human sepsis cases (n = 302). The E. faecium isolated from beef cattle and other food animal sources neither carried vanA/vanB responsible for vancomycin nor possessed gyrA/parC and liaR/liaS gene mutations associated with high-level fluoroquinolone and daptomycin resistance, respectively. A small proportion (7.6%) of human isolates clustered with beef cattle and pig isolates, including a few isolates belonging to the same sequence types ST22 (one beef cattle, one pig, and two human isolates), ST32 (eight beef cattle and one human isolate), and ST327 (two beef cattle and one human isolate), suggesting common origins. This provides further evidence that these clonal lineages may have broader host range but are unrelated to the typical hospital-adapted human strains belonging to clonal complex 17, significant proportions of which contain vanA/vanB and liaR/liaS. Additionally, none of the human isolates belonging to these STs contained resistance genes to WHO critically important antimicrobials. The results confirm that most E. faecium isolated from beef cattle in this study do not pose a significant risk for resistance to critically important antimicrobials and are not associated with current human septic infections.

Correction: Messele et al. Longitudinal Analysis of Antimicrobial Resistance among Enterococcus Species Isolated from Australian Beef Cattle Faeces at Feedlot Entry and Exit. Animals 2022, 12, 2690.

There was an error in the original publication [...].

Meta-Analysis on the Prevalence of Failed Transfer of Passive Immunity in Calves from Pasture-Based Dairy Farms in Australasia.

Monitoring and minimizing the prevalence of failed transfer of passive immunity (FTPI) in dairy replacement calves within the first week of life is crucial for calf health and farm profitability. In this study, a systematic literature search and meta-analysis were conducted on papers reporting the prevalence of FTPI in calves from pasture-based dairy farms in Australia and New Zealand. Two search methods, a "traditional method" and a "search engine method", were conducted to identify published studies on FTPI in Australia and New Zealand. Data from a total of 13,430 calves from eight studies in Australasia were included in the analysis for FTPI within 8 days of birth. The meta-analysis revealed that the average prevalence of FTPI was 33% across the two countries, with the lowest FTPI (9%) in Western Australia and the highest FTPI (59%) in New Zealand. Using farm data from three studies, the average prevalence of FTPI at the farm level in Australasia was 38%, with the lowest prevalence found in a farm in South Australia (6%). In conclusion, the meta-analysis confirmed the need for good management of cows and newborn calves after birth in pasture-based systems to reduce FTPI in calves. Collecting newborn calves from pasture at least twice per day after birth and providing colostrum of sufficient quantity and quality as soon as possible were the best practices for preventing FTPI in Australasian dairy systems.

Associations between Surface and Rectal Temperature Profiles of Low-Birth-Weight Piglets.

The use of infrared cameras to record surface temperature has shown some promise in older pigs, but neonatal piglets are metabolically less mature and experience rapid temperature changes during their first 24 h. The present experiment aimed to compare rectal temperature to surface temperature at the base of the ear, measured using an infrared camera, for piglets of different birth weights. During farrowing, 48 multiparous sows were monitored, and rectal and surface temperatures were recorded for their lower-birth-weight (≤1.2 kg) piglets within 3 min of birth and at 0.25, 0.50, 0.75, 1, 1.25, 1.50, 2, 3, 4, and 24 h. Piglet birth weights were assigned to one of three categories (BWC): BWC1 (≤0.80 kg), BWC2 (0.81 to 1.10 kg), or BWC3 (1.11 to 1.20 kg). Piglet rectal temperatures at 1.25 h after birth were assigned to one of three categories: RC1 (≤32.0 °C), RC2 (32.1 to 35.0 °C), or RC3 (≥35.1 °C). Surface temperatures showed a similar recovery pattern to rectal temperatures in the first 24 h across all piglet birth weights, although large and variable differences seen in the current study militate against surface temperature being an appropriate replacement for neonatal rectal temperature for use in production.

Phylogenetic Analysis of Escherichia coli Isolated from Australian Feedlot Cattle in Comparison to Pig Faecal and Poultry/Human Extraintestinal Isolates.

The similarity of commensal Escherichia coli isolated from healthy cattle to antimicrobial-resistant bacteria causing extraintestinal infections in humans is not fully understood. In this study, we used a bioinformatics approach based on whole genome sequencing data to determine the genetic characteristics and phylogenetic relationships among faecal Escherichia coli isolates from beef cattle (n = 37) from a single feedlot in comparison to previously analysed pig faecal (n = 45), poultry extraintestinal (n = 19), and human extraintestinal E. coli isolates (n = 40) from three previous Australian studies. Most beef cattle and pig isolates belonged to E. coli phylogroups A and B1, whereas most avian and human isolates belonged to B2 and D, although a single human extraintestinal isolate belonged to phylogenetic group A and sequence type (ST) 10. The most common E. coli sequence types (STs) included ST10 for beef cattle, ST361 for pig, ST117 for poultry, and ST73 for human isolates. Extended-spectrum and AmpC ß-lactamase genes were identified in seven out of thirty-seven (18.9%) beef cattle isolates. The most common plasmid replicons identified were IncFIB (AP001918), followed by IncFII, Col156, and IncX1. The results confirm that feedlot cattle isolates examined in this study represent a reduced risk to human and environmental health with regard to being a source of antimicrobial-resistant E. coli of clinical importance.

Neonatal Piglet Temperature Changes: Effect of Intraperitoneal Warm Saline Injection.

Piglets are poor at thermoregulation immediately following birth and take up to 24 h to recover from their initial temperature drop. The present study aimed to determine if providing piglets with a 15 mL intraperitoneal injection of warm (45 °C) saline at birth would improve their internal temperature recovery to 24 h of age, and how the treatment interacted with birth weight (BWC = 1; ≤0.80 kg, BWC = 2; 0.81 kg to 1.10 kg, and BWC = 3; >1.10 kg), rectal temperature at 1.5 h (RC = 1; ≤32.0 °C, RC = 2; 32.10 °C to 35.0 °C, and RC = 3; ≥35.10 °C), and colostrum intake (CI = 1; <200 g and CI = 2, ≥200 g) to affect preweaning survival. Treated BWC1 piglets had improved rectal temperatures from 2 to 24 h. BWC3 piglets who consumed insufficient colostrum also had improved rectal temperature between 1 and 24 h post-birth. Colostrum intake was improved with saline injection in BWC2 piglets of RC1 and RC3 (p < 0.001) and BWC3-RC3 piglets (p < 0.001). Treated BWC1 improved survival to 20 d (p < 0.001). Irrespective of BWC, piglets from all RC had greater survival when injected with saline. The greatest difference was for piglets in RC1, likely due to all BWC1 piglets falling within this category. The results suggest that an intraperitoneal injection of warmed saline is an effective way to improve piglet temperature recovery to 24 h, colostrum intake, and survival in low-birth-weight piglets. These findings will be helpful for producers who have large numbers of low-birth-weight piglets born and are able to provide individual care.

Incidence of Inadequate Transfer of Passive Immunity in Dairy Heifer Calves in South Australia.

The objective of this observational study was to estimate the incidence of inadequate transfer of passive immunity (ITPI) on five pasture-based dairy farms in South Australia. Heifer calf uptake of colostrum was evaluated within the first 1−7 days of age (n = 2638) using a digital refractometer to estimate each calf's serum total protein concentration, as an indicator of colostrum uptake. Results of <51 g/L indicated inadequate transfer of passive immunity (ITPI). The data showed that the incidence of ITPI on the farms was 6.5%, 31.3%, 48.8%, 49.7% and 52.4%. The incidence of ITPI was calculated in relation to the age of the calf at testing and the breed of calf, and no significant differences were found. A significant difference was found in the incidence of ITPI when comparing the calf's first feed after separation from the dam (colostrum versus a colostrum-transition milk mixture). The farm with the lowest incidence of ITPI collected calves twice a day, measured colostrum quality on farm with a Brix refractometer and ensured that each calf received an appropriate amount of high-quality colostrum soon after collection. Further studies are required to establish the risk factors of ITPI in South Australian dairy heifers.

Longitudinal Analysis of Antimicrobial Resistance among Enterococcus Species Isolated from Australian Beef Cattle Faeces at Feedlot Entry and Exit.

Enterococcus faecium are commensal bacteria inhabiting the gastrointestinal tract of animals and humans and an important cause of drug-resistant nosocomial infections. This longitudinal study aimed to determine whether changes in the antimicrobial resistance (AMR) phenotype and genotype occurred among Enterococcus spp. isolated from cattle rectal samples obtained at the entry to and exit from an Australian feedlot. The samples obtained at the feedlot induction yielded enterococci (104/150; 69.3%), speciated as E. hirae (90/104; 86.5%), E. faecium (9/104; 8.7%), E. mundtii (3/104; 2.9%), E. durans, and E. casseliflavus (1/104; 1.0% each). AMR was observed to lincomycin (63/104; 60.6%), daptomycin (26/104; 25.0%), nitrofurantoin (9/104; 8.7%), ciprofloxacin (7/104; 6.7%), tetracycline (5/104; 4.8%), tigecycline (4/104; 3.9%), and quinupristin/dalfopristin (3/104; 2.9%). From the rectal swab samples collected at the abattoir from the same animals (i.e., the feedlot exit), the enterococci recovery was significantly higher (144/150; 96.0%), with a marked shift in species distribution dominated by E. faecium (117/144; 81.3%). However, the prevalence of AMR to individual antimicrobials remained largely static between the entry and exit except for the increased resistance to nitrofurantoin (77/144; 53.5%) and quinupristin/dalfopristin (26/144; 18.1%). Overall, 13 AMR genes were observed among the 62 E. faecium isolates. These included aac(6')Ii, aac(6')-Iid, and ant(6)-Ia (aminoglycosides); eatAv, lnu(G), vat(E), msr(C), and erm(B) (macrolides, lincosamides, and streptogramins); efmA (fluoroquinolones); and tet(45), tet(L), tet(M), and tet(S) (tetracyclines). The results confirm the presence of fluoroquinolone- and streptogramin-resistant enterococci in cattle faeces at the feedlot entry in the absence of antimicrobial selection pressure. E. faecium, exhibiting increased nitrofurantoin resistance, became the dominant Enterococcus spp. during the feeding period.

Phenotypic and Genotypic Analysis of Antimicrobial Resistance in Escherichia coli Recovered from Feedlot Beef Cattle in Australia.

This study investigated the antimicrobial resistance (AMR) profile of fecal Escherichia coli isolates from beef cattle (n = 150) at entry and exit from an Australian feedlot. Sample plating on MacConkey agar and Brilliance ESBL agar differentiated generic from extended-spectrum ß-lactamase (ESBL)-producing E. coli, respectively. Resistance profiles were determined by minimum inhibitory concentration (MIC) testing and further analyzed by whole-genome sequencing (WGS). At entry, the prevalence of antimicrobial resistance to amoxicillin/clavulanic acid, ampicillin, streptomycin, and trimethoprim/sulfamethoxazole was very low (0.7%, each). At the exit, the resistance prevalence was moderate to tetracycline (17.8%) and low to ampicillin (5.4%), streptomycin (4.7%), and sulfisoxazole (3.9%). The most common AMR genes observed in phenotypically resistant isolates were tet(B) (43.2%), aph(3″)-Ib and aph(6)-Id (32.4%), blaTEM-1B, and sul2 (24.3%, each), which are responsible for resistance to tetracyclines, aminoglycosides, ß-lactams, and sulfonamides, respectively. The ESBL-producing E. coli were recovered from one sample (0.7%) obtained at entry and six samples (4.0%) at the exit. The ESBL-producing E. coli harbored blaTEM (29.7%), blaCTX m(13.5%), and blaCMY (5.4%). The resistance phenotypes were highly correlated with resistance genotypes (r ≥ 0.85: p < 0.05). This study demonstrated that E. coli isolated from feedlot beef cattle can harbour AMR genes, but the low incidence of medically important resistance reflected the prudent antimicrobial use in the Australian industry.

Increased feeding frequency prior to farrowing: effects on sow performance.

Reducing the interval between the consumption of the last meal and the start of farrowing is suggested to increase the energy available to sows during farrowing, potentially reducing the farrowing duration and easing piglet births. The present study aimed to examine whether increasing feeding frequency from one to two feeds within standard production hours (0700 to 1500 hours) would produce a difference in farrowing duration and/or stillborn numbers. From entry to farrowing crates (110 ± 1 d gestation) to farrowing (116 ± 1 d gestation), multiparous sows (n = 118) were fed a daily fixed amount of feed either once at 0800 hours or in two meals at 0800 and 1300 hours. Sow weights and backfat depths were recorded on entry and exit from the farrowing crate. Litter size and weight were recorded 24 h after farrowing and on day 21 of lactation. Sows fed twice had a shorter farrowing duration and fewer stillborn piglets than those fed once (2.21 ± 0.56 h vs. 3.25 ± 0.52 h; P = 0.001). The interaction between treatment and farrowing duration showed that sows fed twice have a reduced farrowing duration and had significantly lower stillborn rates than those fed once or those fed twice with longer farrowing durations (P < 0.001). These findings suggest that increasing feeding frequency prior to farrow can reduce the farrowing duration and stillborn numbers in some sows, however, some sows remain with a high stillborn rate regardless of feeding frequency. Piglet average daily gain was greater in once-fed sows, but fewer of these sows remained in the herd at subsequent farrowing. Further, subsequent total born and born alive were higher in twice-fed sows. Feeding sows at a higher frequency can improve farrowing performance in some sows and could increase the longevity of the sow in the herd.

Piglet Morphology: Indicators of Neonatal Viability?

The morphological measures, crown-to-rump length (CR), and abdominal circumference (AC) have been suggested to be as good, if not better, than birth weight for predicting piglet performance. We explored the relationships between CR and AC, and piglet weights at birth and 24 h, to investigate their predictive value for piglet survival. Piglet weight and AC at birth and 24 h, and CR at 24 h were recorded for 373 piglets born to 31 sows. Morphological measures were categorised into two levels for weight and three levels for AC and CR. Further, AC and CR groupings were concatenated to create a new variable (PigProp) to describe the proportionality of piglet morphology. Proportionate piglets had equal CR and AC levels, and disproportionate piglets had contrasting levels. Birth AC was a good predictor of colostrum intake (p < 0.001) when accounting for birth weight, but 24 h weight and PigProp were good indicators of actual colostrum intake (p < 0.001 for both). The significant interaction of colostrum and PigProp showed that within the smaller piglet groups, those who had greater than 200 g of colostrum had higher 24 h weight and survival (p < 0.001 both). As expected, as body weight and colostrum intake increased, so did weight change to d 21 (P = 0.03 and trend at p = 0.1, respectively). A similar pattern was seen with increasing PigProp group (p < 0.001); however, piglets from the disproportionate group 1,3 had the greatest observed weight change (5.15 ± 0.06 kg). Our data show morphological measures may be more predictive of piglet viability in terms of both performance and survival than weight and there may be subgroups that have higher than expected chances of survival.

Effect of body condition score of the dairy cow on the in vitro immune response of peripheral blood mononuclear cells to progesterone stimulation.

Immune function in cows is closely associated with their physical and hormonal conditions. In order to clarify the relationship between the body condition score (BCS) of lactating dairy cows and the immune response to progesterone (P(4)) in vitro, we examined whether lower BCS in dairy cows affects the responsiveness of peripheral blood mononuclear cells (PBMCs) to P(4) added in to culture medium. Forty-two non-pregnant healthy Holstein dairy cows were examined at 61 to 120 days after calving. The cows were divided into the following two groups; Low BCS group (N=20), which had a BCS of less than 2.25, and a Control group (N=22), which had a BCS over 2.75. PBMCs were stimulated with P(4) (1 microg/mL) and/or phytohemagglutinin (PHA), and the levels of cytokine mRNA were analyzed. In the Low BCS group, a significantly lower IFN-gamma level was stimulated by PHA only compared with the Control group. The combination of P(4) and PHA significant decreased the IFN-gamma/IL-4 ratio in the Control group, but this reaction was not found in the Low BCS group. Our data indicated that expression of IFN-gamma mRNA was basically lower in the low BCS dairy cows and that addition of P(4) did not suppress the cellular immune function in these cows. In this study, we observed that P(4) reduced the cellular immune response in the adequate BCS cows, whereas immunosuppression by P(4) was not found in the PBMCs of the low BCS cows, which already had a lower level of immune function.

Comprehensive analysis of machine learning models for prediction of sub-clinical mastitis: Deep Learning and Gradient-Boosted Trees outperform other models.

Sub-clinical bovine mastitis decreases milk quality and production. Moreover, sub-clinical mastitis leads to the use of antibiotics with consequent increased risk of the emergence of antibiotic-resistant bacteria. Therefore, early detection of infected cows is of great importance. The Somatic Cell Count (SCC) day-test used for mastitis surveillance, gives data that fluctuate widely between days, creating questions about its reliability and early prediction power. The recent identification of risk parameters of sub-clinical mastitis based on milking parameters by machine learning models is emerging as a promising new tool to enhance early prediction of mastitis occurrence. To develop the optimal approach for early sub-clinical mastitis prediction, we implemented 2 steps: (1) Finding the best statistical models to accurately link patterns of risk factors to sub-clinical mastitis, and (2) Extending this application from the farms tested to new farms (method generalization). Herein, we applied various machine learning-based prediction systems on a big milking dataset to uncover the best predictive models of sub-clinical mastitis. Data from 364,249 milking instances were collected by an electronic automated in-line monitoring system where milk volume, lactose concentration, electrical conductivity (EC), protein concentration, peak flow and milking time for each sample were measured. To provide a platform for the application of the models developed to other farms, the Z transformation approach was employed. Following this, various prediction systems [Deep Learning (DL), Naïve Bayes, Generalized Liner Model, Logistic Regression, Decision Tree, Gradient-Boosted Tree (GBT) and Random Forest] were applied to the non-transformed milking dataset and to a Z-standardized dataset. ROC (Receiver Operating Characteristics Curve), AUC (Area Under The Curve), and high accuracy demonstrated the high sensitivity of GBT and DL in detecting sub-clinical mastitis. GBT was the most accurate model (accuracy of 84.9%) in prediction of sub-clinical bovine mastitis. These data demonstrate how these models could be applied for prediction of sub-clinical mastitis in multiple bovine herds regardless of the size and sampling techniques.