L.S. Caixeta

Lameness in dairy cattle: A debilitating disease or a disease of debilitated cattle? A cross-sectional study of lameness prevalence and thickness of the digital cushion

Lameness is the most significant challenge for the dairy industry to overcome, given its obvious disruption of animal welfare and severe economic losses. Sole ulcers and white line abscesses are ubiquitous chronic diseases with the highest associated economic losses among all foot lesions. Their underlying causes are still not fully understood. An observational cross-sectional study was carried out to investigate the association between claw horn lesions and the thickness of the digital cushion.The thickness of the digital cushion was evaluated by ultrasonographic examination of the sole at the typical ulcer site. A total of 501 lactating Holstein dairy cows were enrolled in the study. The prevalence of sole ulcers was 4.2 and 27.8% for parity 1 and parity >1, respectively.The prevalence of white line disease was 1.0 and 6.5% for parity 1 and >1, respectively. The prevalence of lameness (visual locomotion score > or = 3) was 19.8 and 48.2% for parity 1 and >1, respectively. The prevalence of sole ulcers and white line diseases was significantly associated with thickness of the digital cushion; cows in the upper quartile of digital cushion thickness had an adjusted prevalence of lameness 15 percentage points lower than the lower quartile. Body condition scores were positively associated with digital cushion thickness.The mean gray value of the sonographic image of the digital cushion had a negative linear association with digital cushion thickness (R2 = 0.14), indicating that the composition of the digital cushion may have changed with its thickness. Furthermore, digital cushion thickness decreased steadily from the first month of lactation and reached a nadir 120 d after parturition.These results support the concept that sole ulcers and white line abscesses are related to contusions within the claw horn capsule and such contusions are a consequence of the lesser capacity of the digital cushion to dampen the pressure exerted by the third phalanx on the soft tissue beneath.

Association between uterine disease and indicators of neutrophil and systemic energy status in lactating Holstein cows.

The objective of this study was to evaluate the association between uterine disease and indicators of neutrophil (PMN) and systemic energy status in dairy cows. Peripheral blood (120 mL) was collected weekly from 84 Holstein cows for PMN isolation and plasma collection from calving until 42 d in milk (DIM). The final analysis included 80 cows. Of those, 20 cows were classified as having metritis (fetid uterine discharge and fever), 15 as having subclinical endometritis (SCE; >or=10% PMN on uterine cytology), and 45 as healthy controls. Plasma haptoglobin concentration was increased only in cows that developed metritis. Neutrophil glycogen content was reduced in cows developing metritis compared with healthy cows on the day of calving and at 7 and 42 DIM. Cows with SCE had lower PMN glycogen content than healthy cows at 7, 28, and 42 DIM. Blood glucose was affected by disease status within parity. Primiparous metritis cows had greater blood glucose concentrations than healthy primiparous cows. Multiparous metritis cows tended to have lower blood glucose concentration than multiparous SCE cows. Cows that developed metritis and SCE had or tended to have greater NEFA and BHBA than healthy cows, mainly around calving. At calving, cows that developed metritis had higher plasma estradiol concentration than healthy cows and greater plasma cortisol than cows that had SCE. Plasma insulin was not affected. Plasma glucagon was increased for SCE cows. Cows that developed uterine disease experienced a greater degree of negative energy balance and had decreased lower intracellular PMN glycogen levels, which could be a major predisposing factor for disease because of decreased availability of oxidative fuels.

Molecular and epidemiological characterization of bovine intrauterine Escherichia coli

Escherichia coli are believed to be associated with postpartum metritis and endometritis but their role in the pathogenesis of both diseases is still undefined. In this study, uterine swabs for E. coli isolation were collected from 374 lactating Holstein cows housed on 4 commercial farms near Ithaca, New York. A total, 125 of 374 cows (33.4%) were positive for E. coli culture. Standard multiplex PCR protocols were used to screen the isolates for the presence of 32 virulence factor genes. Cows that had twin parturition were 4.4 times more likely to have intrauterine E. coli contamination than those that gave birth to single live female calves. Stillborn parturition and birth of single live male calves also increased the odds of intrauterine contamination by E. coli (3.7- and 1.6-fold, respectively) compared with birth of live female calves. Six virulence factors, common to extraintestinal and enteroaggregative E. coli, were found to be associated with metritis and endometritis: fimH, hlyA, cdt, kpsMII, ibeA, and astA. The virulence factor gene fimH was the most prevalent and the most significant: intrauterine E. coli carrying fimH and at least 1 of the other 5 identified virulence factors were pathogenic, and phylogenetic analysis based on the nucleotide sequence of DNA gyrase from 41 such IUEC revealed 2 clades.

Antimicrobial resistance and presence of virulence factor genes in Arcanobacterium pyogenes isolated from the uterus of postpartum dairy cows.

Arcanobacterium pyogenes is considered the most significant bacterium involved in the pathogenesis of metritis in cattle. Infections caused by antimicrobial-resistant bacteria are a great challenge in both human and veterinary medicine. The purpose of this study was to present an overview of antimicrobial resistance in A. pyogenes isolated from the uteruses of postpartum Holstein dairy cows and to identify virulence factors. Seventy-two A. pyogenes isolates were phenotypically characterized for antimicrobial resistance to amoxicillin, ampicillin, ceftiofur, chloramphenicol, florfenicol, oxytetracycline, penicillin, spectinomycin, streptomycin and tetracycline by the broth microdilution method. Presence of virulence factor genes of A. pyogenes was investigated. Isolates exhibited resistance to all antimicrobial agents tested; high levels of resistance were found to amoxicillin (56.9%); ampicillin (86.1%), chloramphenicol (100%), florfenicol (59.7%), oxytetracycline (54.2%), penicillin (86.1%) and tetracycline (50%). Of all isolates, 69 (95.8%) were resistant to at least 2 of the antimicrobial agents tested and multidrug resistance (resistant to at least 3 antimicrobials) was observed in 64 (88.9%) of the A. pyogenes isolates. The major multidrug resistance profile was found for chloramphenicol-ampicillin-penicillin-florfenicol-amoxicillin-tetracycline, which was observed in 21 (29.2%) multidrug resistant isolates. No isolate was resistant to all nine antimicrobial agents tested but four isolates (5.6%) were resistant to eight antimicrobials. The information highlights the need for prudent use of specific antimicrobial agents. All four virulence factor genes occurred in isolates from normal puerperium and clinical metritis; however, the fimA gene was present in significantly higher frequency in isolates from metritis cows.

Effect of an injectable trace mineral supplement containing selenium, copper, zinc, and manganese on the health and production of lactating Holstein cows

The objective of this study was to evaluate the effect of a subcutaneous injection of a multimineral preparation containing 300 mg of zinc, 50mg of manganese, 25mg 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.

Efficacy of formalin, copper sulfate, and a commercial footbath product in the control of digital dermatitis

The objective was to evaluate the relative efficacy of a novel, commercially available disinfectant agent (T-Hexx Dragonhyde HBC, Hydromer, Branchburg, NJ) compared with formalin and copper sulfate. The hypothesis was 2 sided; therefore, the hypothesis was that the new agent would be better or worse compared with the industry gold standard footbath agents, formalin and copper sulfate. The study was conducted in a large commercial dairy farm located near Ithaca, New York, from June 18, 2009 to October 26, 2009. Two identical studies were conducted, the first comparing Dragonhyde (5% solution, twice weekly) and formalin (5% solution, twice weekly) and the second comparing Dragonhyde (5% solution, twice weekly) and copper sulfate (10% solution, twice weekly). The study design was identical for both studies with 4 pens (physically identical), 2 treatments (Dragonhyde vs. formalin and Dragonhyde vs. copper sulfate), 2 periods (crossing over the treatment within pen), and 3 repeated measures (3 observations per cow: enrollment, wk 2, and wk 4). For study 1, 406 cows were enrolled (n=201 formalin and 205 Dragonhyde). For study 2, 356 cows were enrolled (n=189 copper sulfate and 167 Dragonhyde). The adjusted odds of digital dermatitis lesion (DDL) throughout the study period were analyzed by mixed logistic regression model. In study 1, the odds of DDL were 1.36 times higher for the formalin group compared with the Dragonhyde group. In study 2, the data were analyzed by a similar statistical model and the variable treatment did not significantly affect the overall prevalence of DDL. In conclusion, the performance of 3 hoof care products was evaluated and it was concluded that Dragonhyde performed better than formalin and that there was no difference between copper sulfate and Dragonhyde.

Evaluation of cytokine expression by blood monocytes of lactating Holstein cows with or without postpartum uterine disease

Whereas neutrophils are the main phagocytic leukocytes, monocytes and macrophages are actively involved in immunomodulation after infection. Recent studies have demonstrated that neutrophil function is impaired by the state of negative energy balance around parturition, and that cows that develop uterine disease have a greater degree of negative energy balance than healthy cows. The objectives of this study were to compare monocyte gene expression and protein secretion of selected cytokines from calving to 42 d after calving in Holstein cows that did or did not develop uterine disease. Real time quantitative RT-PCR (Tumor necrosis factor-α (TNFα), Interleukin (IL)-1β, IL-6, IL-8 and IL-10) and ELISA (TNFα, IL-1β and IL-8) were used to evaluate cytokine response following in vitro stimulation of blood-derived monocytes with irradiated E. coli. Relative to unstimulated cells, E. coli-stimulated monocytes from cows with metritis had lower gene expression of key pro-inflammatory cytokines than healthy cows from calving to 14 d after calving (TNFα at 0, 7, and 14 d after calving, IL-1β and IL-6 at 7 and 14 d after calving; P < 0.05). There were no significant differences between groups for expression of IL-8 or the anti-inflammatory cytokine IL-10. This was due, in part, to higher gene expression in unstimulated monocytes (TNFα, IL-1β, IL-6 and IL-10) in early lactation from cows with metritis. Expression of mRNA in stimulated cells (relative to housekeeping genes) was lower for TNFα (7 and 14 d postpartum) and for IL-10 (7 and 14 d postpartum) in cows with metritis. Concentration of TNFα was lower in the culture medium of E. coli-stimulated monocytes from cows with metritis than healthy cows at calving and 7 and 21 d after calving (P < 0.05). Circulating cytokine concentrations were not different between groups for IL-8 and were below the limits of detection for TNFα and IL-1β. Cytokine gene expression and production were similar between healthy cows and cows that developed endometritis, diagnosed cytologically at 42 d after calving. We concluded that altered levels of expression and production of pro-inflammatory cytokines postpartum could contribute to impaired inflammatory response and predispose cows to development of metritis.

The effect of claw horn disruption lesions and body condition score at dry-off on survivability, reproductive performance, and milk production in the subsequent lactation

The objective of this study was to evaluate the effects of claw horn disruption lesions (CHDL; sole ulcers and white line disease) and body condition score (BCS) at dry-off on survivability, milk production, and reproductive performance during the subsequent lactation. An observational prospective cohort study was conducted on a large commercial dairy in Cayuga County, New York, from September 2008 until January 2009. A total of 573 cows enrolled at dry-off were scored for body condition and hoof trimmed; digits were visually inspected for the presence of CHDL. The BCS data were recategorized into a 3-level variable BCS group (BCSG), with cows with BCS<3 placed in BCSG 1 (n=113), cows with BCS=3 placed in BCSG 2 (n=254), and cows with BCS>3 placed in BCSG 3 (n=206). Cows in BCSG 2 were 1.35 and 1.02 times more likely to conceive than cows in BCSG 1 and 3, respectively. The cull/death hazard for BCSG 1 cows was 1.55 and 1.47 times higher than for cows in BCSG 2 and BCSG 3, respectively. Milk yield for cows in BCSG 2 (44.6 kg/d, 95% CI 43.4-45.8) was significantly greater than that for cows in BCSG 1 (41.5 kg/d, 95% CI 39.8-43.3). Cows with previous lactation days open<or=91 had 1.6 times higher odds of being classified into BCSG 1 at dry-off; cows with previous lactation mature-equivalent 305-d milk>14,054 kg had a similar 1.6 times higher odds of being classified into BCSG 1. Claw horn disruption lesions were found in 24.4% of the cows (n=140) at dry-off. Cows without CHDL were 1.4 times more likely to conceive than cows with CHDL. Additionally, lesion cows were 1.7 times more likely to die or be culled than nonlesion cows. Absence of CHDL did not have a significant effect on milk yield. These findings highlight the importance of claw health and BCS at the end of lactation on future survival and performance.

Microbial diversity in bovine papillomatous digital dermatitis in Holstein dairy cows from upstate New York

Papillomatous digital dermatitis (PDD) is one of the most prevalent diseases of cattle, adversely affecting the dairy industry by its negative effect on milk production and reproductive performance. Our objective was to use culture-independent methods to determine the microbial diversity in different strata of PDD lesions of three Holstein dairy cows, analyzing whether major differences exist compared to foot skin of three non-infected cows. Both group-specific 16S rRNA gene PCR-denaturing gradient gel electrophoresis and clone library sequencing of broad-range 16S rRNA gene showed differences between the microbial composition of healthy dairy cows and the different strata of the lesion. The predominant bacterial community in the lesion, regardless of the stratum, consisted of 166 specific phylotypes belonging to seven bacterial phyla. Spirochetes (particularly, treponemes) was the most prominent group detected in PDD deep biopsies and was only found in samples from the lesion. Additionally, one phylotype phylogenetically affiliated with uncultured Euryarchaeota was detected in two strata of the lesion. Sequences from healthy foot skin samples revealed 86 specific phylotypes that were affiliated with Firmicutes and Proteobacteria. Our study corroborates the theory that treponemes are involved in PDD disease etiology and suggests, for the first time, the presence of archaeal members in this particular bovine infection.

Susceptibility of Escherichia coli isolated from uteri of postpartum dairy cows to antibiotic and environmental bacteriophages. Part II: In vitro antimicrobial activity evaluation of a bacteriophage cocktail and several antibiotics.

The use of pathogenic-specific antimicrobials, as proposed by bacteriophage therapy, is expected to reduce the incidence of resistance development. Eighty Escherichia coli isolated from uteri of Holstein dairy cows were phenotypically characterized for antimicrobial resistance to ampicillin, ceftiofur, chloramphenicol, florfenicol, spectinomycin, streptomycin, and tetracycline by broth microdilution method. The lytic activity of a bacteriophage cocktail against all isolates was performed by a similar method. Additionally, the effect of different concentrations of antimicrobials and multiplicities of infections (MOI) of the bacteriophage cocktail on E. coli growth curve was measured. Isolates exhibited resistance to ampicillin (33.7%), ceftiofur (1.2%), chloramphenicol (100%), and florfenicol (100%). All strains were resistant to at least 2 of the antimicrobial agents tested; multidrug resistance (>or=3 of 7 antimicrobials tested) was observed in 35% of E. coli isolates. The major multidrug resistance profile was found for ampicillin-chloramphenicol-florfenicol, which was observed in more than 96.4% of the multidrug-resistant isolates. The bacteriophage cocktail preparation showed strong antimicrobial activity against multidrug-resistant E. coli. Multiplicity of infection as low as 10(-4) affected the growth of the E. coli isolates. The ratio of 10 bacteriophage particles per bacterial cell (MOI=10(1)) was efficient in inhibiting at least 50% of all isolates. Higher MOI should be tested in future in vitro studies to establish ratios that completely inhibit bacterial growth during longer periods. All isolates resistant to florfenicol were resistant to chloramphenicol and, because florfenicol was recently introduced into veterinary clinics, this finding suggests that the selection pressure of chloramphenicol, as well as other antimicrobials, may still play a relevant role in the emergence and dissemination of florfenicol resistance in E. coli. The bacteriophage cocktail had a notable capacity to inhibit the in vitro growth of E. coli isolates, and it may be an attractive alternative to conventional treatment of metritis by reducing E. coli in uteri of postpartum dairy cows.