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The aim of this study was to characterize, using metagenomic shotgun DNA sequencing, the intrauterine microbial population and its predicted functional diversity within healthy cows and cows presenting purulent vaginal discharge (PVD). Twenty Holstein dairy cows from a single farm were enrolled in the study at 25 to 35 d postpartum. Purulent vaginal discharge was diagnosed by retrieving and scoring vaginal discharge using the Metricheck device (Simcro, Hamilton, New Zealand). Intrauterine samples for metagenomic analysis were collected by the cytobrush technique from 8 cows diagnosed with PVD and 12 healthy cows. Pair-end sequencing was performed using the Illumina MiSeq platform (Illumina Inc., San Diego, CA). Metagenomic sequences were analyzed using the MG-RAST server (metagenomic rapid annotations using subsystems technology; http://metagenomics.anl. gov/), and the STAMP software (http://kiwi.cs.dal. ca/Software/STAMP) was used to study statistically significant differential abundance of taxonomic and functional features between the 2 metagenomes. Additionally, the total number of bacterial 16S rDNA copies was estimated by real-time PCR. Taxonomic analysis revealed that Bacteroidetes was the most abundant phylum in the uterine microbiota from cows with PVD, and Fusobacteria was almost completely absent in the healthy uterine microbiota. Moreover, species belonging to the genus Trueperella were present only in the uterine microbiota of PVD cows. The increased abundance of Fusobacteria and the unique presence of Trueperella in the PVD cows highlight the important role of these bacteria in the pathogenesis of PVD. Genes encoding cytolethal distending toxin were exclusive to the microbiota of PVD cows. Similarly, genes associated with lipid A modification were present only in samples from PVD cows; such modification is associated with greater resistance to cationic antimicrobial peptides. Conversely, genes encoding bacteriocins and ribosomally antibacterial peptide were exclusively found in the healthy uterine microbiota and dominated by tolerance to colicin E2. No difference was observed in total bacterial load between the 2 microbiotas. This study provides deep insight into the uterine microbial community in health and disease. The observations that the healthy microbiota is tolerant to colicin E2, whereas the uterine microbiota of PVD cows produces cytolethal distending toxins and modifies its lipopolysaccharides suggest that species-intrinsic factors may be more relevant than bacterial abundance to the development of disease or maintenance of health in the dairy cow postpartum uterus. [ABSTRACT FROM AUTHOR]

Metritis is a uterine disease that affects 10 to 30% of all lactating dairy cows and has detrimental effects on reproductive performance, milk production, and survival. Data regarding the identity and abundance of bacterial genes governing traits such as virulence, antibiotic resistance, and stress responses could enable identification of previously unknown agents that play a role in metritis pathogenesis. Moreover, such knowledge could lead to the development of improved treatments or preventive methods. Therefore, the objectives of this study were to characterize the uterine microbial population and to differentiate, for the first time, the microbial functional diversity in cows with metritis versus healthy cows. In addition, we aimed to identify relationships between microbial genes and postpartum uterine health. Uterine swabs were collected from 24 cows within 3 to 12 d in milk; 12 cows were diagnosed with metritis and the other 12 were healthy. Metritis was defined as a watery, reddish or brownish uterine discharge having a fetid smell, and rectal temperature greater than 39.5°C. Cows with a clear and viscous uterine discharge, not fetid or mucopurulent, were classified as healthy. Microbial metagenomic DNA from uterine swab samples was subjected to whole-genome shotgun sequencing on the Illumina MiSeq platform (Illumina Inc., San Diego, CA). The MG-RAST server (metagenomic rapid annotations using subsystems technology; http://metagenomics.anl.gov/) and STAMP software (http://kiwi.cs.dal.ca/Software/STAMP) were used to detect statistically significant differences in the abundance of taxonomic and functional features between the uterine microbial metagenomes of metritic and healthy cows. Our results showed an increased abundance of Fusobacteria and Bacteroidetes in metritic cows, confirming the potential role of those 2 taxa in the pathogenesis of metritis. The MG-RAST analysis revealed a significantly higher abundance of genes for protein transport across the cytoplasmic membrane and type VI bacterial secretion systems in the metritic microbiota. Additionally, genes coding for resistance to acid stress were exclusive to the metritis microbiota, suggesting that microbial resistance to acid stress is important for microbial survival in the infected uterus. On the other hand, genes coding for adhesion molecules, bacteriocins, and antibacterial peptides were significantly associated with the uterine microbiota of healthy cows, as was tolerance to colicin E2. [ABSTRACT FROM AUTHOR]

We investigated the microbiota found in the vaginas of Holstein dairy cows during the transition period and described the differences in bacterial composition and total bacterial load (TBL) associated with disease and fertility. Vaginal swabs were collected at -7, 0, 3, and 7 d relative to parturition from 111 dairy cows housed on a commercial dairy farm near Ithaca, New York. Microbiota were characterized by next-generation DNA sequencing of the bacterial 16S rRNA gene, and TBL was determined by real-time quantitative PCR. We applied repeated-measures ANOVA to evaluate the associations of uterine disease and related risk factors with the microbiota and TBL. We estimated phylum-specific bacterial load by multiplying the TBL by the relative abundance of each phylum observed in the metagenomics results. We confirmed the validity of this approach for estimating bacterial load by enumerating the number of bacteria in an artificial sample mixed in vitro and in clinical and healthy vaginal samples. Phyla associated with uterine disease and related risk factors were Proteobacteria, Fusobacteria, and Bacteroidetes. Cows with retained placenta and healthy cows had similar TBL at the day of parturition, but at d 7 postpartum, cows with retained placenta showed a significantly higher TBL, mainly driven by higher estimated loads of Fusobacteria and Bacteroidetes. Cows diagnosed with metritis had a significantly higher estimated load of Proteobacteria at d -7 and at calving and higher estimated loads of Fusobacteria in the postpartum samples. Additionally, the estimated load of Bacteroidetes at d 7 postpartum was higher for cows diagnosed with endometritis at 35 days in milk. Higher estimated loads of Fusobacteria and Bacteroidetes were also evident in cows with postpartum fever, in primiparous cows, in cows with assisted parturition, and in cows that gave birth to twins. Our findings demonstrated that microbiota composition and TBL were associated with known periparturient risk factors of uterine diseases and reproductive failure, including parity, assisted parturition, and retained fetal membranes. [ABSTRACT FROM AUTHOR]

Postpartum uterine diseases of dairy cows compromise animal welfare and may result in early removal from the herd or impaired reproductive performance. The relationship between poor immune status around calving and uterine diseases is well established; however, that between natural antibodies (NAb) and uterine health has not yet been studied. The objective of this study was to evaluate the association of circulating NAb levels around parturition with puerperal metritis, clinical endometritis, and the intrauterine presence of the Escherichia coli virulence factor FimH. One hundred six pregnant heifers were enrolled; NAb in serum samples collected at 30 ± 3 d prepartum and at 2 ± 1 and 35 ± 3 d in milk (DIM) were measured by ELISA. Puerperal metritis was defined as the presence of fetid, watery, red-brown uterine discharge and rectal temperature >39.5°C at 6 ± 1 DIM. Clinical endometritis was defined as presence of pus in the uterine lavage sample collected at 35 ± 3 DIM. The intrauterine presence of the fimH gene at 2 ± 1 DIM was evaluated by PCR. The overall optical density (wavelength of 650 nm) of ELISA-detected serum NAb was lower for cows diagnosed with puerperal metritis than for cows that did not have puerperal metritis. Additionally, cows diagnosed with clinical endometritis tended to have lower levels of NAb than did cows without clinical endometritis. Finally, FimH-positive cows had lower overall levels of serum NAb compared with FimH-negative cows. In conclusion, NAb detected in serum around parturition was associated with uterine health of dairy cows. [ABSTRACT FROM AUTHOR]

The objective of this study was to evaluate the relationship between selected minerals' serum levels, energy metabolites, oxidative stress indicators, IL-8 and haptoglobin levels, and the potential for uterine diseases. Additionally, we investigated the effect of injectable trace mineral supplementation (ITMS) on metabolism, immune function, and animal health under field conditions involving a dairy herd with high milk production. The study was conducted in 1 dairy farm located near Ithaca, New York, with 270 multiparous cows were enrolled from October 3, 2012 until January 10, 2013. Cows were randomly allocated into 1 of 2 treatments groups: ITMS or control. Cows randomly assigned to the ITMS group received 2 injections of trace minerals at 230 and 260 d of gestation; each injection contained 300 mg of Zn, 50 mg of Mn, 25 mg of Se, and 75 mg of Cu. Retained placenta (RP) and metritis were diagnosed and treated by trained farm personnel. Clinical endometritis evaluation was performed by the investigators. Blood mineral levels, plasma nonesterified fatty acids and serum β-hydroxybutyrate concentrations, plasma IL-8 concentrations, serum haptoglobin concentration, and serum superoxidase dismutase and plasma glutathione peroxidase activities were measured at various time points before and after calving. Four groups of mixed general linear models were fitted to the data using MIXED procedure of SAS. Injectable trace mineral-supplemented cows had increased serum concentration of Cu, Se, and Zn. Conversely, ITMS did not affect energy metabolites or immune and oxidative stress parameters. Serum concentration of Ca, Cu, K, Mg, Mo, Ps, Pt, Se, and Zn varied according to days relative to parturition. Cows with RP had reduced serum concentrations of Ca, Mg, Mo, and Zn when compared with cows without RP. Cows affected with metritis had significantly lower serum concentrations of Ca, Mo, soluble P, total P, Se, and Zn than nonaffected cows. Serum concentration of Ca, Cu, Mo, and Zn were reduced in cows diagnosed with endometritis in comparison to nonaffected ones. [ABSTRACT FROM AUTHOR]

Raw milk and colostrum can harbor dangerous micro-organisms that can pose serious health risks for animals and humans. According to the USDA, more than 58% of calves in the United States are fed unpasteurized milk. The aim of this study was to evaluate the effect of UV light on reduction of bacteria in milk and colostrum, and on colostrum IgG. A pilot-scale UV light continuous (UVC) flow-through unit (45 J/cm²) was used to treat milk and colostrum. Colostrum and sterile whole milk were inoculated with Listeria innocua, Mycobacterium smegmatis, Salmonella serovar Typhimurium, Escherichia coli, Staphylococcus aureus, Streptococcus agalactiae, and Acinetobacter baumannii before being treated with UVC. During UVC treatment, samples were collected at 5 time points and bacteria were enumerated using selective media. The effect of UVC on IgG was evaluated using raw colostrum from a nearby dairy farm without the addition of bacteria. For each colostrum batch, samples were collected at several different time points and IgG was measured using ELISA. The UVC treatment of milk resulted in a significant final count (log cfu/mL) reduction of Listeria monocytogenes (3.2 ± 0.3 log cfu/mL reduction), Salmonella spp. (3.7 ± 0.2 log cfu/mL reduction), Escherichia coli (2.8 ± 0.2 log cfu/mL reduction), Staph. aureus (3.4 ± 0.3 log cfu/mL reduction), Streptococcus spp. (3.4 ± 0.4 log cfu/mL reduction), and A. baumannii (2.8 ± 0.2 log cfu/mL reduction). The UVC treatment of milk did not result in a significant final count (log cfu/mL) reduction for M. smegmatis (1.8 ± 0.5 log cfu/mL reduction). The UVC treatment of colostrum was significantly associated with a final reduction of bacterial count (log cfu/ mL) of Listeria spp. (1.4 ± 0.3 log cfu/mL reduction), Salmonella spp. (1.0 ± 0.2 log cfu/mL reduction), and Acinetobacter spp. (1.1 ± 0.3 log cfu/mL reduction), but not of E. coli (0.5 ± 0.3 log cfu/mL reduction), Strep. agalactiae (0.8 ± 0.2 log cfu/mL reduction), and Staph. aureus (0.4 ± 0.2 log cfu/mL reduction). The UVC treatment of colostrum significantly decreased the IgG concentration, with an observed final mean IgG reduction of approximately 50%. Development of new methods to reduce bacterial contaminants in colostrum must take into consideration the barriers imposed by its opacity and organic components, and account for the incidental damage to IgG caused by manipulating colostrum. [ABSTRACT FROM AUTHOR]

The objective of this study was to evaluate the effect of a subcutaneous injection of a multimineral preparation containing 300 mg of zinc, 50 mg of manganese, 25 mg of selenium, and 75 mg of copper at 230 and 260 days of gestation and 35 days postpartum, on the health, milk production and reproductive performance of lactating Holstein cows. A randomized field trial was conducted on three large commercial dairy farms located near Ithaca, New York, USA, with 1416 cows enrolled. All cows were housed and offered a total mixed ration consisting of approximately 55% forage and 45% concentrate on a dry matter basis of the diet, which supplied 2-6 times the NRC requirements for the supplemented elements. Dry cows and pregnant heifers were blocked by parity and randomly allocated to one of two treatments: Trace mineral supplemented (TMS) or control. For multiparous cows, subcutaneous TMS significantly decreased linear somatic cell count scores (normalized data) as compared to control cows. The incidence of subclinical mastitis for TMS and control cows was 10.4% and 8.0%, respectively (P = 0.005). The main effect of treatment on clinical mastitis was not significant but the interaction of treatment and parity was significant. For primiparous cows, the incidence of clinical mastitis was 11.8% and 15.6% for control and TMS cows, respectively (P = 0.33); for multiparous cows, the incidence of clinical mastitis for control and TMS cows was 25.4% and 19.7%, respectively (P = 0.03). Additionally, control cows had increased odds of stillbirth and endometritis (odds ratios 1.69 and 1.30, respectively). The incidence of endometritis was 34.2% and 28.6% for control and TMS cows, respectively (P = 0.039) but treatment had no effect on reproductive performance, milk production or other health traits. Further research is required to confirm these findings and to establish whether the response seen in this study was related to the supplementation of a particular mineral. [ABSTRACT FROM AUTHOR]

The aim of this study was to evaluate the effects of different physical treatments of bovine colostrum and hospital milk on milk bacteriology, immunoglobulin G (IgG) and lactoferrin concentrations, calf serum IgC concentrations and calf health, growth and survivability. Pooled colostrum samples (n = 297) were heat treated (HTC; 63°C for 60 mm), exposed to ultraviolet light (UVC: 45 J/cm²) or untreated ('raw', RC). Hospital milk (n = 712) was subjected to high temperature short time pasteurization (HTST; 72°C for 15 s), ultraviolet light irradiation (UVH; 45 J/cm²) or was untreated. Neonatal Holstein heifer calves (n = 875) were randomly enrolled (309 HTC, 285 UVC, 281 RC) and block randomized (by colostrum treatment) into hospital milk treatments HTST (n = 449) or UVH (n = 426). HTC was more effective than UVC and HTST was more effective than UVH in reducing bacterial counts. IgG and lactoferrin concentrations were significantly lower in HTC and UVC than in RC. Lactoferrin concentrations were significantly lower in HTST than in UVH or untreated hospital milk. There were no significant differences in serum IgG concentrations among calves fed HTC, UVC or RC. Colostrum and hospital milk treatments did not have any significant effect on calf body weight gain, survivability, or frequency of diarrhea or pneumonia. [ABSTRACT FROM AUTHOR]

The objective of this study was to evaluate the effects of intrauterine administration of 50 g of ultrapure mannose or a bacteriophage cocktail and the presence of Escherichia coli and Arcanobacterium pyogenes in the uterine lumen on uterine health and reproductive performance of lactating dairy cows. The study was conducted on a commercial dairy farm located near Ithaca, New York, from May 4 to January 20, 2011, and 597 cows were enrolled. The cows were divided randomly into 3 treatment groups, and treatments were administered at 2 ± 1 d in milk (DIM). Treatment 1 consisted of intrauterine administration of 50 g of ultrapure mannose powder divided in 4 pills; treatment 2 consisted of intrauterine administration of a bacteriophage cocktail that included 4 different phages in one pill, with a dose of approximately 107 plaqueforming units; and treatment 3 consisted of intrauterine administration of one empty pill (control). Intrauterine fluid swabs were collected on day of treatment and at 10 ± 3 DIM; uterine lavage samples were collected at 35 ± 3 DIM. Swabs and uterine lavage samples were cultured for E. coli and A. pyogenes. The intrauterine administration of mannose and bacteriophage did not affect uterine health, reproduction performance, or outcome of intrauterine secretion cultures for E. coli and A. pyogenes. Prevalence of intrauterine E. coli at 2 ± 1 DIM and A. pyogenes at 2 ± 1, 10 ± 3, and 35 ± 3 were affected by retained placenta. Additionally, prevalence of intrauterine E. coli at 10 ± 3 and A. pyogenes at 35 ± 3 DIM were associated with metritis, and cows that were diagnosed with clinical endometritis at 35 ± 3 DIM had greater prevalence of intrauterine E. coli at 2 ± 1 DIM and A. pyogenes at 35 ± 3 DIM. Furthermore, cows positive for E. coli at 2 ± 1 and 10 ± 3 DIM and for A. pyogenes at 10 ± 3 DIM had 1.63, 2.34, and 1.54 increased odds of having metritis. Cows positive for A. pyogenes at 35 ± 3 DIM and for E. coli at 2 ± 1 DIM had 19.8 and 2.66 higher odds of being diagnosed with clinical endometritis, respectively. Additionally, cows negative for E. coli at 2 ± 1 DIM were 1.39 times more likely to conceive than cows positive cows for E. coli. [ABSTRACT FROM AUTHOR]

Bone Marrow Transplant (BMT) is a therapy used to treat patients with hematological diseases. The success of the transplant relies on a HLA match between host and donor. The HLA is located in the Major Histocompatibility Complex in the 6p12.3 region of the chromosome 6. The HLA gene products are involved in the immunomodulation of the immune response due to their function of presenting peptides to the T cells. The HLA genes are the most polymorphic in humans and the most relevant genetic marker for clinical transplants and are largely used in population studies. The knowledge of the HLA-A, HLA-B and HLA-DR haplotype frequencies of bone marrow donors is an important tool when a patient needs an identical HLA donor, and there are few population studies similar to this in Brazil. The HLA typing was performed in the LIGH of the UFPR by the PCR-SSP technique. The most common haplotypes among the population studied were HLA-A*01B*08DR*03, HLA-A*29B*44DR*07 and HLA-A*03B*07DR*15. The search of a Brazilian patient for an identical HLA donor is usually hopeless and the understanding of the HLA frequencies permits a real foreknowledge of the success of this search. Success depends on the eventual registration of the perfect donor in the national centers of bone marrow donation. Aiming to increase the perspectives of patients who need a BMT, the evaluation of the HLA frequencies and the enhancement of the national registrations of bone marrow donors are crucial for the accomplishment of this objective.

The new allele might have arisen from HLA-A*66:01 through a point mutation at codon 182.1 (ACG→GCG) resulting in a non-conservative change from threonine to alanine. [ABSTRACT FROM AUTHOR]

The new alleles, HLA-B*40:125 and HLA-B*40:129, present a mismatch at codon 103.1 (G → C) and three mismatches at codons 9.1 (C → T), 11.1 (T → G) and 12.1 (G → A). [ABSTRACT FROM AUTHOR]

Abstract Background The causal association between persistent human papillomavirus (HPV) infection and cervical cancer has been established, but the mechanisms that favor HPV persistence in cervical cells are still unknown. The diminished capability of the immune system to control and resolve HPV infection is one of several hypotheses. The tolerogenic protein HLA-G has shown aberrant expression in a variety of cancers, which has been suggested as a mechanism for tumor escape from immunosurveillance. In the present study we evaluate the role of epigenetic modification (promoter de-methylation) of the HLA-G gene on susceptibility to HPV infection and development of high-grade cervical lesions. Methods A case–control study was carried out in Curitiba, Brazil, between February and June 2010. A total of 789 women aged 15–47 years were recruited: 510 controls with normal cervical cytology, and 279 cases with histologically confirmed cervical intraepithelial neoplasia grade 2 (CIN2, N = 150) or grade 3 (CIN3, N = 129). All women were administered a questionnaire by interview, which collected information on demographic and lifestyle factors, and a cervical sample was collected. HPV DNA detection was performed by GP5+/GP6+ primer-mediated PCR. HPV-positive samples were genotyped by multiplex PCR. A pilot analysis of HLA-G promoter methylation was carried out in a subset of the study population (96 cases and 76 controls) by pyrosequencing. HLA-G methylation and HPV infection status of cases and controls were compared, and confounding factors were computed by t Student and non-parametric Wilcoxon tests. Comparison of HLA-G methylation between cases and controls was assessed by the Bonferroni correction. The association of HLA-G methylation with CIN2/3 was evaluated by logistic regression. Results HPV prevalence was 19.6% in controls and 94.3% in CIN2/3 cases. HPV16, 31, 33, 35 and 18 were the most prevalent types. Methylation analysis of seven CpGs in the HLA-G promoter did not reveal any spontaneous de-methylation events in CIN2/3 cases (mean proportion of methylation: 75.8%) with respect to controls (mean 73.7%; odds ratio 1.01, 95% confidence interval 0.96, 1.07). Conclusions This study did not support the hypothesis that spontaneous de-methylation events in the HLA-G promoter play a primary role in promoting escape from immunosurveillance in the development of precancerous cervical lesions.

The objective of this study was to evaluate the effect of 2 subcutaneous injections of a multimineral preparation, each containing 60 mg of zinc, 10 mg of manganese, 5 mg of selenium, and 15 mg of copper at 3 and 30 d after birth on immunity, health, and growth of dairy calves during the preweaning period. The study was conducted in upstate New York in 2 commercial dairy farms. A total of 790 Holstein heifer calves were randomly allocated at birth into 1 of 2 treatments: trace mineral supplement (TMS) treated or control. Blood samples were collected at 3, 14, and 35 d after birth to evaluate glutathione peroxidase (GPx) activity, superoxide dismutase (SOD) activity, haptoglobin, and neutrophil and monocyte function. Incidence of diseases and average daily gain was evaluated in the first 50 d of life. At 14 d of life, TMS-treated calves had increased neutrophil activity compared with control calves. Moreover, TMS-treated calves had greater GPx activity on d 14 after birth than control calves. The TMS treatment reduced the incidence of diarrhea (TMS = 41.7% vs. control = 49.7%) and combined incidence of pneumonia or otitis or both (TMS = 41.7% vs. control = 49.1%). Additionally, GPx was greater for calves diagnosed with otitis at d 35 after birth. However, calves diagnosed with pneumonia had decreased GPx activity at d 35 after birth. Serum SOD and haptoglobin concentrations were not affected by treatment or disease. Moreover, no effects were observed on average daily gain and survivability between TMS-treated and control calves during the preweaning period. Supplementation with trace minerals at 3 and 30 d of life increased neutrophil function and GPx activity and reduced the incidence of health disorders. [ABSTRACT FROM AUTHOR]

The objectives were (1) to evaluate the association of uterine lavage sample optical density (ULSOD) with uterine health, and (2) to estimate and evaluate a threshold value that will maximize the accuracy of ULSOD as a diagnostic tool for clinical endometritis. The study enrolled 1,742 cows from 3 dairy farms located near Ithaca, New York. The samples were collected at 35 ± 3 d in milk (DIM) by using low-volume uterine lavage. Cows with a purulent or mucopurulent secretion in the sample were diagnosed with clinical endometritis, whereas a subgroup of all studied cows was examined for cytological evidence of inflammation by endometrial cytology. Data for ULSOD measured at different wavelengths (200, 352, 620, 790, 860, and 960 nm) were available for 554 cows; all 1,742 cows had data for ULSOD measured at 620 nm (ULSOD620). Incidences of clinical endometritis, metritis, and retained placenta were 10, 15.2, and 5.6%, respectively. The ULSOD620 was associated with clinical endometritis. Receiver operating characteristic (ROC) analysis of the accuracy of optical density in the detection of clinical endometritis was conducted for ULSOD measured at different wavelengths; ULSOD620 was selected for further analysis because it presented the best ROC curve to detect clinical endometritis. The recommended threshold for ULSOD620 ROC was 0.058, where the sensitivity and specificity were 76.3 and 78.3%, respectively. The ROC analysis of the accuracy of optical density in the detection of endometritis defined as a percentage of neutrophils in the uterine lavage samples higher than 18% was conducted for ULSOD620. The recommended threshold was 0.059, where the sensitivity and specificity were 100 and 82.2%, respectively. Cows with ULSOD620 ≤0.058 were 1.21 times more likely to conceive than cows with ULSOD620 >0.058; moreover, the median calving-to-conception interval for cows that had ULSOD620 ≤0.058 was 122 d compared with 148 d for cows that had ULSOD620 >0.058. Cows that were positive for Arcanobacterium pyogenes, diagnosed with metritis, or had retained placenta had 4.0, 1.4, and 1.7 times higher odds of having ULSOD620 >0.058, respectively. Cows with ULSOD620 >0.058 had a higher percentage of neutrophils in the uterine lavage samples than cows with ULSOD620 ≤0.058. Uterine lavage sample optical density measured at 620 nm can be used as an objective indicator of uterine health in dairy cows, principally for clinical endometritis. [ABSTRACT FROM AUTHOR]

Diarrhea in calves has a significant effect on the dairy industry. A common management practice for preventing or decreasing the effects of such disease in preweaned calves is by the use of antimicrobials in milk or milk replacer. In this study, Escherichia coli antimicrobial resistance in fecal samples collected from calves 2 to 8 d of age that had or had not received antimicrobials in the milk and that had or had not signs of diarrhea by inspection of fecal consistency were investigated. Specifically, resistance of E. coli isolates to individual antimicrobials, multiresistance patterns, and presence of virulence factors were analyzed. Escherichia coli isolates were tested for susceptibility to 12 antimicrobials by use of a Kirby-Bauer disk diffusion assay. The study was conducted at 3 farms, 1 administering growth-promoting antimicrobials (GPA) in the milk and 2 not using GPA in the milk (NGPA). All isolates were susceptible to ciprofloxacin and cefepime. From the total isolates tested, 84% (n = 251) were resistant to at least 2 antimicrobials and 81% (n = 251) were resistant to 3 or more antimicrobials. When antimicrobial resistance was compared between GPA and NGPA, it was observed that the GPA group had higher odds of antimicrobial resistance for most of the individual antimicrobials tested. No significant correlation of virulence factors in GPA or NGPA and diarrheic or non-diarrheic (control) fecal samples was found. Of the 32 virulence factors evaluated, 21 were detected in the study population; the incidence of only 1 virulence factor was statistically significant in each of the diarrheic status (diarrheic or non-diarrheic) and treatment status (NGPA or GPA) groups. Phylogenetic analysis based on the nucleotide sequence of the DNA gyrase gene (gyrB) from 31 fecal E. coli isolates revealed 3 main clades. [ABSTRACT FROM AUTHOR]

The main objective of this study was to investigate the association between digital cushion thickness and sole temperature measured by infrared thermography. Data were collected from 216 lactating Holstein cows at 4 to 10 d in milk (DIM). Cows were locomotion scored and sole temperature was measured after claw trimming (a minimum delay of 3 min was allowed for the hoof to cool) using an infrared thermography camera. Temperature was measured at the typical ulcer site of the lateral digit of the left hind foot. Immediately after the thermographic image was obtained, the thickness of the digital cushion was measured by ultrasonography. Rumen fluid samples were collected with a stomach tube and sample pH was measured immediately after collection. Additionally, a blood sample was obtained and used for measurements of serum concentrations of β-hydroxybutyrate (BHBA), nonesterified fatty acids (NEFA), and haptoglobin. To evaluate the associations of digital cushion thickness with sole temperature, a linear regression model was built using the GLIMMIX procedure in SAS software (SAS Institute Inc., Cary, NC). Sole temperature was the response variable, and digital cushion thickness quartiles, locomotion score group, rumen fluid pH, rumen fluid sample volume, environmental temperature, age in days, and serum levels of NEFA, BHBA, and haptoglobin were fitted in the model. Only significant variables were retained in the final model. Simple linear regression scatter plots were used to illustrate associations between sole temperature (measured by infrared thermography at the typical ulcer site) and environmental temperature and between NEFA and BHBA serum levels and haptoglobin. One-way ANOVA was used to compare rumen fluid pH for different locomotion score groups and for different digital cushion quartiles. Results from the multivariable linear regression model showed that sole temperature increased as locomotion scores increased and decreased as digital cushion thickness increased. These results were adjusted for environmental temperature, which was significantly associated with sole temperature. Serum levels of NEFA, BHBA, and haptoglobin were not associated with sole temperature. However, significant correlations existed between serum levels of NEFA and haptoglobin and between serum levels of BHBA and haptoglobin. Rumen fluid pH was not associated with either locomotion score or digital cushion thickness. In conclusion, we show here that digital cushion thickness was associated with sole temperature in cows at 4 to 10 DIM. [ABSTRACT FROM AUTHOR]

The objective of this study was to evaluate the effect of subcutaneous supplementation of 300 mg of zinc, 50 mg of manganese, 25 mg of selenium, and 75 mg of copper on peripheral blood leukocyte activity and serum β-hydroxybutyrate (BHBA) concentrations at 10 ± 2 days in milk (DIM), and on serum Superoxide dismutase (SOD) activity during the transition period and subsequent lactation of multiparous Holstein cows. A total of 250 multiparous cows were randomly allocated into one of two treatments groups, namely, trace mineral supplemented (TMS) or control Cows in the TMS group were injected at 230 and 260 days of gestation, and 35 days postpartum. Serum SOD activity was measured at enrollment, and 10, 60 and 100 DIM. Serum BHBA concentration and leukocyte function were assessed at 10 DIM. Overall serum SOD activity for TMS and control was 16.01 and 12.71 U/mL, respectively. The interaction between treatment and time of serum collection was significant. Additionally, overall serum SOD activity was 12.85 and 14.78 U/mL for cows diagnosed with mastitis and unaffected cows, respectively. Treatment did not affect leukocyte function. For parity >2, TMS cows had lower serum BHBA concentrations than control cows; BHBA concentrations were 0.41 and 0.27 mmol/L for control and TMS cows, respectively. In conclusion, cows diagnosed with mastitis had decreased serum SOD activity, and trace mineral supplementation increased serum SOD activity although leukocyte function was not affected by supplementation. [ABSTRACT FROM AUTHOR]

The objective of this study was to isolate bacteriophages from environmental samples of 2 large commercial dairy farms using Escherichia coli isolated from the uteri of postpartum Holstein dairy cows as hosts. A total of 11 bacteriophage preparations were isolated from manure systems of commercial dairy farms and characterized for in vitro antimicrobial activity. In addition, a total of 57 E. coli uterine isolates from 5 dairy cows were phylogenetically grouped by triplex PCR. Each E. coli bacterial host from the uterus was inoculated with their respective bacteriophage preparation at several different multiplicities of infections (MOI) to determine minimum inhibitory MOI. The effect of a single dose (MOI = 10²) of bacteriophage on the growth curve of all 57 E. coli isolates was assessed using a microplate technique. Furthermore, genetic diversity within and between the different bacteriophage preparations was assessed by bacteriophage purification followed by DNA extraction, restriction, and agarose gel electrophoresis. Phylogenetic grouping based on triplex PCR showed that all isolates of E. coli belonged to phylogroup B1. Bacterial growth was completely inhibited at considerably low MOI, and the effect of a single dose (MOI = 10²) of bacteriophage preparations on the growth curve of all 57 E. coli isolates showed that all bacteriophage preparations significantly decreased the growth rate of the isolates. Bacteriophage preparation 1230-10 had the greatest antimicrobial activity and completely inhibited the growth of 71.7% (n = 57) of the isolates. The combined action of bacteriophage preparations 1230-10, 6375-10, 2540-4, and 6547-2, each at MOI = 102, had the broadest spectrum of action and completely inhibited the growth (final optical density at 600 nm ≤0.1) of 80% of the E. coli isolates and considerably inhibited the growth (final optical density at 600 nm ≤0.2) of 90% of the E. coli isolates. Restriction profile analysis demonstrated that all 4 phage preparations contained bacteriophages that were genetically distinct from each other according to the banding pattern of the fragments. The combination of several different bacteriophages can improve the spectrum of action, and the results of this study suggest that bacteriophages 1230-10, 6375-10, 2540-4, and 6547-2 should be used in combination as a cocktail. [ABSTRACT FROM AUTHOR]

The HLA-A*23:140 allele differs from HLA-A*23:01:01 by one nucleotide substitution (G > A), position 1968 in exon 5., (© 2024 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)