Supplementation of rumen-protected lysine during the close-up period improves vaginal discharge clearance in Holstein dairy cows.

We aimed to evaluate the effects of rumen-protected lysine (RPL) supplementation during the close-up period on uterine involution and the resumption of ovarian function in dairy cows. Fifty-two multiparous Holstein cows were categorized based on parity and expected calving date and randomly assigned to the RPL or control (CON) groups. The RPL group received 80 g of RPL daily from day 21 before the expected calving date until parturition. Blood samples were obtained twice weekly from pre-supplementation to 6 weeks postpartum. The onset of luteal activity postpartum was determined via ultrasonography twice weekly for up to 6 weeks postpartum. Uterine involution was tracked at 3 and 5 weeks postpartum through the vaginal discharge score, percentage of polymorphonuclear cells (PMN) in endometrial cytology samples, presence of intrauterine fluid, and gravid horn diameter via ultrasonography. Before supplementation, the RPL group showed amino acid imbalance, which was improved by RPL supplementation. There were no significant differences in the onset of luteal activity, percentage of PMN, intrauterine fluid, or the diameter of the uterine horn between the two groups. The vaginal discharge score in the RPL group decreased from 3 to 5 weeks postpartum, whereas that in the CON groups did not decrease. The number of cows with clinical endometritis was lower in the RPL group. Overall, RPL supplementation during the close-up period enhanced vaginal discharge clearance, potentially averting clinical endometritis, but did not affect the first ovulation in dairy cows.

Comparison between intrauterine application of an antibiotic and an herbal product to treat clinical endometritis in dairy cattle - A randomized multicentre field study.

Uterine diseases are main indications for antibiotic use in dairy cows. To test a non-antibiotic treatment option, we compared the effect of an intrauterine cephapirin (Metricure®; cefapirin benzathin 500 mg per dosis; CEPH) with an intrauterine applied herbal product (25 ml of EucaComp® PlantaVet containing alcoholic extracts of Calendula officinalis L., Mellissa officinalis L., Origanum majorana L. and Eucalyptus globulus Labill. (EUC)) on the clinical cure of endometritis. Examinations of 816 cows between 21 and 35 days after calving were performed and cases of clinical endometritis (n = 169) were included. Diagnosis based on a scoring system for vaginal discharge. Study animals were randomly assigned to one of two treatment groups and treated immediately. After excluding animals with incomplete datasets, 136 cows (EUC: n = 61; CEPH: n = 75) remained for the final analysis. In total, 64% (EUC: 61%, CEPH: 67%) of analysed endometritis cases were considered as clinically cured 14 ± 2 days after the first treatment, 15% stayed uncured after the application of a maximum of two consecutive treatments, leading to an overall clinical cure rate of 85% (EUC: 82%, CEPH: 88%). No statistically difference in clinical cure rates could be observed between both treatment groups nor 14 ± 2 days after the first treatment (p = 0.956) neither regarding the overall cure rate (p = 0.923). In conclusion, the clinical cure of dairy cows' endometritis after the intrauterine application of the herbal product was non-inferior to the intrauterine application of the antibiotic cephapirin. These results could contribute to reduce the antimicrobial use in the daily veterinary routine treatment of endometritis.

Alternatives to conventional antibiotics for the prevention and treatment of commonly occurring diseases in feedlot cattle.

Antibiotic-resistant bacteria are a problem in human medicine. The development of antibiotic resistance in bacteria in feedlot cattle could have negative effects on their health and welfare and there is a theoretical possibility of transmission of antibiotic-resistant bacteria from food animals to humans. Alternatives to conventional antibiotics in feedlot health management could reduce the selective pressure for the development of antibiotic resistance. This review assesses the evidence supporting potential alternatives to conventional antibiotics in the prevention and treatment of diseases in feedlot cattle, including nitric oxide, plant extracts, supplemental yeast or yeast products, bacterial probiotics, organic acids, bacteriophages and non-specific immunostimulants. Further research is warranted with lactate utilising bacteria, the organic acid malate, bacteriophages and the non-specific immunostimulants ß-1,3 glucan and those based on pox viruses. However, none of the alternatives to conventional antibiotics investigated in this review have sufficient supporting evidence to date to justify their use with feedlot cattle. Frequently, statistically weak results and studies without negative controls are cited as support for similar studies. The health and welfare of feedlot cattle are dependent on the use of products that have robust supporting data to ensure efficacy and to avoid adverse outcomes.

Utilizing intramammary Melaleuca alternifolia as an organic internal sealant for dry-off therapy in Murrah buffaloes.

The effects of intramammary dry cow therapy based on the administration of 5% Melaleuca alternifolia tea tree essential oil (TTO) as an internal teat sealant to Murrah cows were evaluated. A longitudinal prospective and retrospective negative control study was performed using 12 buffaloes from a total of 20 Murrah buffaloes on an organic farm, with the cow used as a control for herself. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) for treatments with pure oil (TTO) and medication containing 5% TTO (O5) were determined. The buffaloes were clinically examined, and the teats were evaluated using thermography and ultrasound. Udder health was monitored during the first 100 days in milk (DIM) using milk somatic cell count (SCC) and California mastitis test (CMT). Laboratory tests against standard strains Staphylococcus aureus ATCC®25,923™, Escherichia coli ATCC®25,922™, and wild bacterial strains showed maximum MIC values of 50 µL/mL for the TTO and O5 treatments. One wild-type S. aureus strain showed no MBC. No adverse effects were observed after the intramammary application of TTO. The CMT and SCC values were similar (P > 0.05) for all observations. The medication containing 5% TTO was effective in vitro and compatible with the intramammary tissue in vivo of Murrah buffaloes. TTO was safe, not inducing inflammatory processes or other modifications of the teat detectable by thermography or ultrasound. It was able to protect buffaloes during the dry period under field conditions, demonstrating potential use as a teat sealant for organic farms.

Efficacy of hydroethanolic extract of Randia aculeata seed against the southern cattle tick Rhipicephalus (Boophilus) microplus (Acari: Ixodidae) on naturally infested cattle under field conditions.

The cattle tick Rhipicephalus (Boophilus) microplus is a major problem of concern for cattle industry in tropical and subtropical areas. Control of cattle tick is based mainly on the use of chemical acaricides, which has contributed to the emerging problem of selection of resistant tick lineages. Plants have been used as an alternative to conventional acaricidal drugs. On the other hand, the acaricidal activity of hydroethanolic extract of Randia aculeata seed (EHRA) has been demonstrated against R. microplus under laboratory conditions. However, the utility of EHRA seed as a potential acaricidal needs to be determined under field conditions. For this reason, the aim of this study was to evaluate the efficacy of the EHRA against R. microplus sprayed on naturally infested calves, determine the effect of the EHRA seed on acetylcholinesterase activity in R. microplus larval and identify the chemical composition of EHRA. Forty-five male calves were divided in three groups and treated with: G1 water; G2 EHRA 20% w/v and G3 coumaphos 0.2% v/v. Acetylcholinesterase (AChE) activity in R. microplus larvae was determined by a colorimetric assay. The chemical composition of EHRA was accessed through HPLC/MS. Significantly fewer ticks were observed after 24 h on the treated group compared to control group. EHRA significantly inhibited in vitro AChE activity in R. microplus at all tested concentrations. Chlorogenic acid, vanillinic acid, p-coumaric acid, caffeic acid. rutin, quercetin, (-)-epicatechin, 4-hydroxybenzoic acid, quercetin, vanillin, 2,4-dimethoxy-6-methylbenzoic acid, scopoletin and ferulic acid were identified in the extract. The results provided new data for the elucidation of the mechanisms of EHRA acaricide action and to further evaluate the use as a new alternative control agent against R. microplus under in vivo conditions.

Successful treatment with streptomycin of genital leptospirosis in naturally infected cows under field conditions.

The single-dose protocol of streptomycin treatment has been recommended to treat renal leptospirosis in bovines. However, treating genital infection remains a challenge. Recently, a protocol using three doses of streptomycin demonstrated effectiveness in the genital clearance of experimentally infected ewes. Therefore, the present study aimed to apply this three-dose protocol for genital infection treatment in naturally infected cows under field conditions. Thirty beef cows were diagnosed as positive by lipL32-PCR in their genital samples. Nucleotide sequences (n = 10) characterized them as Leptospira interrogans sg Sejroe, genetically related to Hardjoprajitno strains. After molecular diagnosis, 13 cows received a single dose of 25 mg/kg streptomycin. The other 17 cows were submitted to the three-dose protocol. The successful treatment rate of genital infection on the single streptomycin dose was 7/13 (53.8%), while the cows that received the three doses 16/17 were negative (94.1% of efficacy). Based on those results, we conclude that the standard treatment preconized for renal infection is not adequate for genital infection, and the three-dose protocol was successful in eliminating the carrier status of genital leptospirosis.

Network meta-analysis of the therapeutic effects of various antibiotics on footrot in sheep and cattle.

The present network meta-analysis was performed to compare the effects of antibiotics used in treating footrot in some ruminants and to rank these antibiotics based on their efficacy. Data of 14 eligible studies consisting of 5622 affected animals was included in the analysis. A Bayesian method and Markov Chain Monte Carlo (MCMC) simulations were utilized to analyze data. The estimated results were reported in the form of odds ratios (ORs) with 95% credible intervals (CrI). The Surface Under the Cumulative Ranking Curve (SUCRA) was used to rank antibiotics. Network meta-regressions (NMRs) were conducted to examine the influence of sample sizes, treatment duration, route of administration, and species of animals (sheep and cattle) on the overall outcome. The results indicated that gamithromycin impact on curing footrot was superior to other antibiotics and Lincomycin and oxytetracycline were ranked second and third. The difference between the impact of gamithromycin and amoxicillin (OR = 14.76, CrI: 1.07-193.49) and enrofloxacin (OR = 20.21, CrI: 1.57-229.25) on footrot was significant. There was a significant difference between the effect of oxytetracycline and enrofloxacin (OR = 5.24, CrI: 1.14-23.74) on footrot. The NMR performed based on species of animals fitted data better than network meta-analysis, suggesting erythromycin as the best third antibiotic instead of oxytetracycline. Egger's regression test and the shape of the funnel plot showed no publication bias among included studies. In conclusion, gamithromycin was associated with the highest curing rate benefit when used to treat footrot, followed by lincomycin and oxytetracycline/erythromycin. Among all evaluated antibiotics, enrofloxacin showed the lowest effects on footrot.

Response of cows with osteomalacia grazing sub-tropical native pastures to phosphorus supplementation with loose mineral mix or feed blocks.

Osteomalacia outbreaks often occur in cattle grazing native pastures in regions with endemic phosphorous (P) deficiency. This study evaluated the responses of two groups of cows, initially with clinical signs of chronic P deficiency, to P supplements (100 g P/kg) offered ad libitum for 13 weeks as a loose mineral mix (LMM group) or the same mineral mix offered as blocks (BMM group). Half of the cows in each group were categorized as 'with' or 'without' severe osteopenia according to a test that depended on the resistance to penetration of a needle through the left lateral process of the L4-L5 lumbar vertebra. The groups grazed two paddocks that were switched each 3 weeks. The liveweight, supplement intakes, and the P-concentrations in soil, forage, blood, and external cortical bone (ECB) of the ribs were measured. The bicarbonate-extractable P in soil was 3.5 mg/kg. The mean of total P in forage (0.95 g/kg/DM), inorganic P in serum (iP, 0.96 mmol/L), and total P in the ECB of the ribs (85 mg/mL) at the beginning of the experiment were all low and consistent with severe chronic P deficiency. The P supplementation allowed clinical recovery in 18/20 cows with their serum and ECB P and calcium approaching normal values and in the two remaining cows the only sign was abnormal gait. Cows consumed more of the LMM than BMM supplement (means 8.3 and 6.6 g P/day, respectively). After 13 weeks cows initially classified as 'with severe osteopenia' and supplemented with LMM had higher (P < 0.05) final liveweight (difference = 21.6 kg), iP (difference = 0.74 mmol/L), bone Ca (difference = 65.7 mg/mL) and bone P (difference = 26.5 mg/mL) concentrations and lower (P < 0.01) final serum Ca/iP ratio (difference = -0.65) than cows with severe osteopenia but supplemented with BMM. The treatment of severe P deficiency cows grazing P deficient sub-tropical grasslands by P supplementation for 13 weeks was more effective with LMM than BMM.

Antibiofilm action using water-soluble tetra-cationic porphyrin and antibacterial photodynamic therapy against Moraxella spp. from cattle.

Infectious bovine keratoconjunctivitis (IBK) is the most important eye disease in ruminants worldwide. Moraxella bovis and Moraxella bovoculi can form biofilm and are frequently isolated from affected animals. Antimicrobials are used worldwide to treat clinical cases of IBK, although they have limited success in clearing the infection. Therefore, photodynamic therapy using porphyrins as photosensitizing molecules is an alternative method to eliminate microorganisms, including biofilms. We evaluated the antibacterial activity of a zinc(II) metalloporphyrin (ZnTMeP) against M. bovis and M. bovoculi biofilms since this compound can efficiently inactivate planktonic Moraxella spp. This study was carried out with two reference strains of Moraxella spp. (M. bovis: ATCC® 10900 and M. bovoculli: ATCC® BAA1259). The antibacterial activity of 4.0 µM of the ZnTMeP porphyrin was evaluated on forming and consolidate biofilms with three 30-min cycles of white-light exposure for three days. The ZnTMeP porphyrin reduced M. bovis and M. bovoculi biofilm formation. In addition, ZnTMeP partially destroyed consolidated M. bovoculi biofilms in the second white-light irradiation cycle, although the porphyrin had no effect against the consolidated biofilm of M. bovis. Despite the biofilm still not being completely inactivated, our findings are promising and encourage further experiments using the phototherapy protocol.

Use of a product based on wood vinegar of Eucalyptus clone I144 used in the control of bovine mastitis.

Bovine mastitis is one of the most frequent diseases in dairy cattle worldwide. The use of antiseptics in milking, if properly used, can lead to a reduction in potentially pathogenic microorganisms and their transmission between herds. Several medicinal plants have antiseptic potential, including eucalyptus (Eucalyptus spp.). Therefore, the objective of this study was to evaluate the effectiveness of wood vinegar from Eucalyptus urograndis clone GG I144 (EU) as an antiseptic in vitro and in vivo; in addition, to its cytotoxicity and antimicrobial resistance. Fifteen bovines were used, lactating females 3-6 years of age and divided into three groups of five animals each. The wood vinegar was placed in the teats of the animal for 28 days and collections of cellular debris were performed every 7 days. At the Veterinary Microbiology Laboratory (LAMIV) of UFERSA, the samples were processed and serial dilution was performed in Petri plates with plate count agar (PCA) at 37 °C. Cytotoxicity was verified based on morphological alterations and metabolic activity. Morphological changes were not observed in all cells incubated with 1 % pyroligneous extract. The in vitro data demonstrated antimicrobial activity against S. aureus, S. agalactiae, Salmonella, E. coli and P. aeruginosa. The bacteria of the genus Staphylococcus and Corynebacterium were resistant to penicillin (PEN), rifampicin (RIF), nitrofurantoin (NIT), erythromycin (ERI), and ciprofloxacin (CIP). The extract was used in vivo in the post-dipping of dairy cows, which reduced the microbiological load present in the mammary glands from 4.74 to 2.54 CFU, indicating its future use as an antiseptic.

[Phytotherapeutic treatments of gynecological diseases and fertility disorders in cattle - a veterinary historical analysis]. / Phytotherapeutische Behandlungen gynäkologischer Erkrankungen und Fruchtbarkeitsstörungen von Rindern ­ eine veterinärhistorische Analyse.

INTRODUCTION: Fertility problems are the main reasons for culling dairy cows. Diseases of the female genital tract are also often the cause of antibiotic or hormonal treatments in bovine practices. The use of medicinal plants could expand the available therapeutics. The aim of the work was to analyze historical literature before the introduction of antibiotics in veterinary reproductive medicine. Five books in German language, published in Germany and Switzerland between 1878 and 1921, and one handwritten therapy booklet by the rural veterinarian Carl Ammann-Honegger (1879-1960) were systematically examined regarding the descriptions (AW) on gynecological diseases. The herbal and additional ingredients of the recipe, the target animal species, the type of administration and the indication were recorded in detail for each AW. The six literature sources contained a total of 103 AW (79 administered orally, 13 locally, and 11 both orally and locally). Almost two thirds of the AW (61) were based on a mixture of different plants (two to seven plants), and one third of the AW (31) on a single plant. A total of 55 plants were recorded. The most frequently mentioned medical plants were plants of the genus Juniperus (J. communis L. (19 AW), J. sabina L. (13 AW)) and Linum usitatissimum L. (18 AW), Matricaria chamomilla L. (13 AW) and Gentiana lutea L. (12 AW). The treatment of the Retentio secundinarum was the most frequently mentioned indication (44 AW), followed by parturition preparation (17 AW) and endometritis treatment (15 AW). The most frequently recorded plants can be divided based on their ingredients and their effect into (a) energy- and protein-rich forage plants, (b) generally appetizing, digestive- and metabolism-enhancing plants, (c) medical plants with a specific gynecological organotrophic effect and (d) according to current knowledge, predominantly toxic plants. Besides the antimicrobial active immunity to defence against bacterial infections, a stable barrier funcion of the endometrium contributes to uterine health. The plants classified under (a) - (c) have at least the potential for a positive effect on the immune system and the endometrial barrier function and thus contribute indirectly to the uterine health.

INTRODUCTION: Les problèmes de fertilité sont les principales raisons de la mise à la réforme des vaches laitières. Les maladies de l'appareil génital femelle sont également souvent à l'origine de traitements antibiotiques ou hormonaux dans les pratiques bovines. L'utilisation de plantes médicinales pourrait élargir les thérapeutiques disponibles. L'objectif de ce travail était d'analyser la littérature historique avant l'introduction des antibiotiques en médecine de la reproduction vétérinaire. Cinq livres en langue allemande, publiés en Allemagne et en Suisse entre 1878 et 1921, et un livret thérapeutique écrit à la main par le vétérinaire rural Carl Ammann-Honegger (1879­1960) ont été systématiquement examinés en ce qui concerne les descriptions d'utilisation (AW) sur les maladies gynécologiques. Les herbes et les ingrédients supplémentaires de la recette, l'espèce animale cible, le type d'administration et l'indication ont été enregistrés en détail pour chaque AW. Les six sources de littérature contenaient un total de 103 AW (79 administrées par voie orale, 13 par voie locale et 11 par voie orale et locale). Près de deux tiers des AW (61) étaient basés sur un mélange de différentes plantes (deux à sept plantes), et un tiers des AW (31) sur une seule plante. Au total, 55 plantes ont été enregistrées. Les plantes médicinales les plus fréquemment mentionnées étaient les plantes du genre Juniperus (J. communis L. (19 AW), J. sabina L. (13 AW)) et Linum usitatissimum L. (18 AW), Matricaria chamomilla L. (13 AW) et Gentiana lutea L. (12 AW). Le traitement du Retentio secundinarum a été l>indication la plus fréquemment mentionnée (44 AW), suivie de la préparation à la parturition (17 AW) et du traitement de l>endométrite (15 AW). Les plantes les plus fréquemment enregistrées peuvent être divisées, en fonction de leurs composants et de leurs effets, en (a) plantes fourragères riches en énergie et en protéines, (b) plantes généralement appétissantes, favorisant la digestion et le métabolisme, (c) plantes médicinales ayant un effet organotropique gynécologique spécifique et (d) selon les connaissances actuelles, plantes principalement toxiques. Outre l'effet antimicrobien, une capacité de défense stable de l'endomètre contribue à la santé utérine. Les plantes classées sous (a) - (c) ont au moins le potentiel d'un effet positif sur le système immunitaire et la fonction de défense de l'endomètre et contribuent ainsi indirectement à la santé utérine.

Evaluation of the efficacy of cow colostrum in the treatment and its effect on serum proteomes in calves with cryptosporidiosis.

This study aimed to investigate the effect of using high-quality colostrum in addition to paromomycin on the treatment outcomes and serum proteomes of calves naturally affected by cryptosporidiosis. Thirty Holstein calves infected with only Cryptosporidium spp. were divided into three equal groups. Calves in the PC group received paromomycin orally at a dose of 100 mg/kg once daily for 5 days. Calves in PCOL and PBCOL groups received 250 ml colostrum 3 h after feeding twice a day for 3 days. The PBCOL group was also given 6 g of sodium bicarbonate 15 min before colostrum administration. While the fecal scores of all calves were evaluated daily for 10 days from the initiation of the treatment, fecal oocyst counts were determined on the 0, 1, 2, 3, 5, 7, and 10th days. Brix%, total protein (TP), immunoglobulin G (IgG), gamma-glutamyltransferase (GGT) levels, and proteomic analyses were performed on the 0 and 3rd days' sera. Considering pretreatment values, fecal scores (8th, 2nd, and 2nd day), and fecal oocyst counts (10th, 3rd, and 2nd day) improved in a significantly (p < 0.05) shorter time in the colostrum groups than in the control group. By serum proteomic analysis, 99, 93, and 83 proteomes were detected in PC, PCOL, and PBCOL groups, respectively. Although the significant changes in any protein in Group PC were absent, significant changes were observed in Alpha-1B-glycoprotein (A1BG), Zinc transporterZIP11 (S39AB), Cathelicidin-1 (CTHL1), Actin_ cytoplasmic-1 (ACTB), and Apolipoprotein A-IV (APOA4) proteins in Group PCOL and Alpha-1-antiproteinase (A1AT), Serum amyloid A protein (SAA), Actin-cytoplasmic-2 (ACTG), Protein HP-20 homolog (HP20) proteins in Group PBCOL with colostral treatment, which indicated that the use of colostrum had an effect on calf serum proteomes. The more pronounced healing and shorter clinical improvement time in the colostrum groups especially colostrum with sodium bicarbonate revealed that these proteomes have positive effects in the treatment with their systemic and local effects in the intestines.

The use of herbal treatments as alternatives to control uterine diseases in dairy cows.

Uterine diseases are a common gynecological issue that affects dairy cows during the postpartum period. The treatment of uterine diseases is mainly managed through antimicrobials; however, the problem of antibiotic residues, increased antimicrobial resistance of pathogens, and the abuse of antibiotics causes serious problems in the dairy chain around the world, which can affect the efficiency of conventional medicines. In these regards, alternative treatment strategies are needed to reduce the use of antibiotics in dairy production. This review primarily provides an overview that focuses on various alternative sources and methodologies, including plants extracts and essential oils, for the control of bovine uterine diseases, and it presents the advantages and limitations related to the use of these herbal therapies.

Limitations of bacterial culture, viral PCR, and tulathromycin susceptibility from upper respiratory tract samples in predicting clinical outcome of tulathromycin control or treatment of bovine respiratory disease in high-risk feeder heifers.

A cross-sectional prospective cohort study including 1026 heifers administered tulathromycin due to high risk of clinical signs of bovine respiratory disease (BRD), measured poor association between BRD clinical outcomes and results of bacterial culture and tulathromycin susceptibility from BRD isolates of deep nasopharyngeal swabs (DNS) and adequate association with viral polymerase chain reaction (PCR) results from nasal swabs. Isolation rates from DNS collected on day-0 and at 1st BRD-treatment respectively were: Mannheimia haemolytica (10.9% & 34.1%); Pasteurella multocida (10.4% & 7.4%); Mycoplasma bovis (1.0% & 36.6%); and Histophilus somni (0.7% & 6.3%). Prevalence of BRD viral nucleic acid on nasal swabs collected exclusively at 1st BRD-treatment were: bovine parainfluenza virus type-3 (bPIV-3) 34.1%; bovine viral diarrhea virus (BVDV) 26.3%; bovine herpes virus type-1 (BHV-1) 10.8%; and bovine respiratory syncytial virus (BRSV) 54.1%. Increased relative risk, at 95% confidence intervals, of 1st BRD-treatment failure was associated with positive viral PCR results: BVDV 1.39 (1.17-1.66), bPIV-3 1.26 (1.06-1.51), BHV-1 1.52 (1.25-1.83), and BRSV 1.35 (1.11-1.63) from nasal swabs collected at 1st BRD-treatment and culture of M. haemolytica 1.23 (1.00-1.51) from DNS collected at day-0. However, in this population of high-risk feeder heifers, the predictive values of susceptible and resistant isolates had inadequate association with BRD clinical outcome. These results indicate, that using tulathromycin susceptibility testing of isolates of M. haemolytica or P. multocida from DNS collected on arrival or at 1st BRD-treatment to evaluate tulathromycin clinical efficacy, is unreliable.

Characterization and comparison of the microbiomes and resistomes of colostrum from selectively treated dry cows.

Professionals in animal agriculture promote prudent use of antimicrobials to address public and animal health concerns, such as reduction of antimicrobial residues and antimicrobial resistance (AMR) in products. Few studies evaluate the effect of selective dry-cow therapy on preservation of the milk microbiome or the profile of AMR genes (the resistome) present at freshening. Our objectives were to characterize and compare the microbiomes and resistomes in the colostrum of cows with low somatic cell count that were treated or not treated with intramammary cephapirin benzathine at dry-off. From a larger parent study, cows on a New York dairy farm eligible for dry-off and with histories of somatic cell counts ≤200,000 cells/mL were enrolled to this study (n = 307). Cows were randomly assigned to receive an intramammary antimicrobial and external teat sealant (ABXTS) or sealant only (TS) at dry-off. Composite colostrum samples taken within 4 h of freshening, and quarter milk samples taken at 1 to 7 d in milk were subjected to aerobic culture. The DNA extraction was performed on colostrum from cows with culture-negative samples (ABXTS = 43; TS = 33). The DNA from cows of the same treatment group and parity were pooled (26 pools; ABXTS = 12; TS = 14) for 16S rRNA metagenomic sequencing. Separately, the resistome was captured using a custom RNA bait library for target-enriched sequencing. Sequencing reads were aligned to taxonomic and AMR databases to characterize the microbiome and resistome, respectively. The R statistical program was used to tabulate abundances and to analyze differences in diversity measures and in composition between treatment groups. In the microbiome, the most abundant phyla were Firmicutes (68%), Proteobacteria (23%), Actinobacteria (4%), and Bacteroidetes (3%). Shannon and richness diversity means were 0.93 and 14.7 for ABXTS and 0.94 and 13.1 for TS, respectively. Using analysis of similarities (ANOSIM), overall microbiome composition was found to be similar between treatment groups at the phylum (ANOSIM R = 0.005), class (ANOSIM R = 0.04), and order (ANOSIM R = -0.04) levels. In the resistome, we identified AMR gene accessions associated with 14 unique mechanisms of resistance across 9 different drug classes in 14 samples (TS = 9, ABXTS = 5). The majority of reads aligned to gene accessions that confer resistance to aminoglycoside (TS = ABXTS each 35% abundance), tetracycline (TS = 22%, ABXTS = 54%), and ß-lactam classes (TS = 15%, ABXTS = 12%). Shannon diversity means for AMR class and mechanism, respectively, were 0.66 and 0.69 for TS and 0.19 and 0.19 for ABXTS. Resistome richness diversity means for class and mechanism were 3.1 and 3.4 for TS and 1.4 and 1.4 for ABXTS. Finally, resistome composition was similar between groups at the class (ANOSIM R = -0.20) and mechanism levels (ANOSIM R = 0.01). Although no critical differences were found between treatment groups regarding their microbiome or resistome composition in this study, a larger sample size, deeper sequencing, and additional methodology is needed to identify more subtle differences, such as between lower-abundance features.

Antimicrobial activity of polyphenols extracted from Thai medical plants on extended-spectrum beta-lactamase-producing Escherichia coli isolates from healthy dairy cows.

Escherichia coli producing extended-spectrum beta-lactamase (ESBL) are antimicrobial- -resistant Enterobacteriaceae important in the livestock production sector, especially dairy cows because these antimicrobial-resistant bacteria can be transferred to consumers via consumption. If antimicrobials are continually used in dairy farms, this may result in antimicrobial resistance. Therefore, investigation of antimicrobial resistance and finding new alternative methods for inhibiting ESBL-producing E. coli is essential. Hence, the aim of this study was to examine the ability of selected antimicrobials to inhibit E. coli ATCC 25922, control bacteria and ESBL-producing E. coli isolated from dairy farms. We also investigated the capacity of polyphenol extract from 10 varieties of medicinal plants to inhibit ESBL-producing E. coli using a broth microdilution method. It was found that control bacteria were susceptible to all antimicrobial agents, i.e., ampicillin, cefotaxime, ciprofloxacin, chloramphenicol, gentamycin, imipenem, nalidixic acid, tetracycline, and sulfamethoxazole/ trimethoprim. However, ESBL-producing E. coli exhibited both susceptibility and resistance to selected antimicrobials. The polyphenol extracted from Psidium guajava Linn at the lowest concentration was 4.5 mg/mL, which could inhibit control bacteria, but at the same concentration could not inhibit ESBL-producing E. coli. These phenomena indicated that ESBL-producing E. coli had both susceptibility and resistance to antimicrobials. Polyphenol, which could inhibit non-resistant E. coli, could not inhibit ESBL-producing E. coli.

Use of a natural herbal-based feed additive containing isoquinoline alkaloids in newborn calves with cryptosporidiosis.

Cryptosporidium infections are one of the most prevalent causes of diarrhea in calves and considered to be one of the major sources of economic loss in livestock production. A global trend is currently underway, in identifying natural and sustainable alternatives to support animal husbandry and production. Isoquinoline alkaloids are known for their anti-inflammatory, antimicrobial, and antioxidant properties in the promotion of gut health. Thus, an experiment was designed to evaluate the effects of natural, herbal-based feed isoquinoline alkaloids to support calves experimentally inoculated with Cryptosporidium parvum. Twenty-six calves were randomly divided into control (CN) (n = 13) and treatment (SG) (n = 13) groups. The SG group received 5 g of feed additive in every milk feeding from 1 to 21 days of age. The CN group received milk without any additives. All calves were orally inoculated on the third day of life with 1 × 106Cryptosporidium parvum oocysts. The animals were evaluated daily, from 3 to 30 days of age, for the occurrence, duration, and intensity of diarrhea. Calves with a base deficit of ≥ 9 mEq/L were hydrated to aid recovery. The SG calves showed a higher average weight gain between 14 and 21 days of age, without mortality and with reduced intensity and duration of diarrhea. In contrast, calves in the CN group showed more serious acid-base disorders, required more hydration support, and had a mortality rate of 15.4 %. These results showed that calves supplemented with isoquinoline alkaloids had decreased intensity and duration of symptoms, reduced requirement for supportive therapy, and prevented mortality among animals.

Genome-Wide Association Study Reveals Genetic Markers for Antimicrobial Resistance in Mycoplasma bovis.

Mycoplasma bovis causes many health and welfare problems in cattle. Due to the absence of clear insights regarding transmission dynamics and the lack of a registered vaccine in Europe, control of an outbreak depends mainly on antimicrobial therapy. Unfortunately, antimicrobial susceptibility testing (AST) is usually not performed, because it is time-consuming and no standard protocol or clinical breakpoints are available. Fast identification of genetic markers associated with acquired resistance may at least partly resolve former issues. Therefore, the aims of this study were to implement a first genome-wide association study (GWAS) approach to identify genetic markers linked to antimicrobial resistance (AMR) in M. bovis using rapid long-read sequencing and to evaluate different epidemiological cutoff (ECOFF) thresholds. High-quality genomes of 100 M. bovis isolates were generated by Nanopore sequencing, and isolates were categorized as wild-type or non-wild-type isolates based on MIC testing results. Subsequently, a k-mer-based GWAS analysis was performed to link genotypes with phenotypes based on different ECOFF thresholds. This resulted in potential genetic markers for macrolides (gamithromycin and tylosin) (23S rRNA gene and 50S ribosomal unit) and enrofloxacin (GyrA and ParC). Also, for tilmicosin and the tetracyclines, previously described mutations in both 23S rRNA alleles and in one or both 16S rRNA alleles were observed. In addition, two new 16S rRNA mutations were possibly associated with gentamicin resistance. In conclusion, this study shows the potential of quick high-quality Nanopore sequencing and GWAS analysis in the evaluation of phenotypic ECOFF thresholds and the rapid identification of M. bovis strains with acquired resistance. IMPORTANCE Mycoplasma bovis is a leading cause of pneumonia but also causes other clinical signs in cattle. Since no effective vaccine is available, current M. bovis outbreak treatment relies primarily on the use of antimicrobials. However, M. bovis is naturally resistant to different antimicrobials, and acquired resistance against macrolides and fluoroquinolones is frequently described. Therefore, AST is important to provide appropriate and rapid antimicrobial treatment in the framework of AMR and to prevent the disease from spreading and/or becoming chronic. Unfortunately, phenotypic AST is time-consuming and, due to the lack of clinical breakpoints, the interpretation of AST in M. bovis is limited to the use of ECOFF values. Therefore, the objective of this study was to identify known and potentially new genetic markers linked to AMR phenotypes of M. bovis isolates, exploiting the power of a GWAS approach. For this, we used high-quality and complete Nanopore-sequenced M. bovis genomes of 100 isolates.

Ubericin K, a New Pore-Forming Bacteriocin Targeting mannose-PTS.

Bovine mastitis infection in dairy cattle is a significant economic burden for the dairy industry globally. To reduce the use of antibiotics in treatment of clinical mastitis, new alternative treatment options are needed. Antimicrobial peptides from bacteria, also known as bacteriocins, are potential alternatives for combating mastitis pathogens. In search of novel bacteriocins against mastitis pathogens, we screened samples of Norwegian bovine raw milk and found a Streptococcus uberis strain with potent antimicrobial activity toward Enterococcus, Streptococcus, Listeria, and Lactococcus. Whole-genome sequencing of the strain revealed a multibacteriocin gene cluster encoding one class IIb bacteriocin, two class IId bacteriocins, in addition to a three-component regulatory system and a dedicated ABC transporter. Isolation and purification of the antimicrobial activity from culture supernatants resulted in the detection of a 6.3-kDa mass peak by matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry, a mass corresponding to the predicted size of one of the class IId bacteriocins. The identification of this bacteriocin, called ubericin K, was further confirmed by in vitro protein synthesis, which showed the same inhibitory spectrum as the purified antimicrobial compound. Ubericin K shows highest sequence similarity to the class IId bacteriocins bovicin 255, lactococcin A, and garvieacin Q. We found that ubericin K uses the sugar transporter mannose phosphotransferase (PTS) as a target receptor. Further, by using the pHlourin sensor system to detect intracellular pH changes due to leakage across the membrane, ubericin K was shown to be a pore former, killing target cells by membrane disruption. IMPORTANCE Bacterial infections in dairy cows are a major burden to farmers worldwide because infected cows require expensive treatments and produce less milk. Today, infected cows are treated with antibiotics, a practice that is becoming less effective due to antibiotic resistance. Compounds other than antibiotics also exist that kill bacteria causing infections in cows; these compounds, known as bacteriocins, are natural products produced by other bacteria in the environment. In this work, we discover a new bacteriocin that we call ubericin K, which kills several species of bacteria known to cause infections in dairy cows. We also use in vitro synthesis as a novel method for rapidly characterizing bacteriocins directly from genomic data, which could be useful for other researchers. We believe that ubericin K and the methods described in this work will aid in the transition away from antibiotics in the dairy industry.

The effect of injectable vitamin C and road transit duration on inflammation, muscle fatigue, and performance in pre-conditioned beef steer calves.

This study examined the effects of injectable vitamin C (VC) before transport and duration of transit on feedlot performance, inflammation, and muscle fatigue in cattle. One hundred thirty-two Angus-cross steers (393 ± 4 kg) were stratified by body weight (BW) to a 2 × 2 factorial of intramuscular injection (INJ; 20 mL/steer): VC (250 mg sodium ascorbate/mL) or saline (SAL) and road transit duration (DUR): 18 h (18-h; 1,770 km) or 8 h (8-h; 727 km). On day 0, steers were weighed and given INJ of VC or SAL immediately before transport. Upon return (day 1), BW and blood were collected before steers returned to pens equipped with GrowSafe bunks. Steers were weighed on days 0, 1, 7, 15, 30, 31, 54, and 55. Data were analyzed via ProcMixed of SAS (experimental unit = steer; 32 to 34 steers/treatment) with fixed effects of INJ, DUR, and the interaction. Blood was collected on days -5, 1, 2, 3, and 7 (n = 9 steers/treatment); blood parameters were analyzed as repeated measures with the repeated effect of day. Area under the curve (AUC) for plasma ferric reducing antioxidant power (FRAP) was calculated using R. Final BW was greater for 8 h compared to 18 h (P = 0.05) with no effect of INJ or interaction (P ≥ 0.51). Dry matter intake (DMI) from days 1 to 7 was greater for VC-8, intermediate for VC-18 and SAL-18, and least for SAL-8 (P = 0.02). Overall, DMI tended to be greatest for SAL-18, intermediate for VC-18 and VC-8, and least for SAL-8 (P = 0.08). Days 7 to 31 gain:feed (G:F) was greatest for VC-18 compared to other treatments (INJ × DUR, P = 0.05), and there was no effect of treatment on overall G:F (P ≥ 0. 19). There was no INJ or INJ × DAY (P ≥ 0.17) effect on serum lactate, haptoglobin, or non-esterified fatty acid. However, these blood parameters were greater on day 1 for 18 h compared to 8 h, and both treatments returned to near baseline by day 3 (DUR × DAY, P < 0.01). Plasma ascorbate concentrations on day 1 were greater for VC compared to SAL and returned to baseline by day 2 (INJ × DAY, P < 0.01). Plasma FRAP AUC from days -5 to 3 was greatest for VC-18, intermediate for VC-8 and SAL-8, and least for SAL-18 (INJ × DAY, P = 0.02). This suggests an antioxidant prior to long-haul transit positively influenced antioxidant capacity; however, VC did not improve overall post-transit performance. Although longer transit duration increased indicators of muscle fatigue and inflammation, post-transit performance was not appreciably different between transit durations.