Comparative diagnoses of respiratory disease in preweaned dairy calves using sequential thoracic ultrasonography and clinical respiratory scoring.

AIMS: Bovine respiratory disease (BRD) has serious impacts on dairy production and animal welfare. It is most commonly diagnosed based on clinical respiratory signs (CRS), but in recent years, thoracic ultrasonography (TUS) has emerged as a diagnostic tool with improved sensitivity and specificity. This study aimed to assess the alignment of BRD diagnoses based on a Clinical Respiratory Scoring Chart (CRSC) and weekly TUS findings throughout the progression of BRD of variable severity in preweaned Holstein dairy heifers. METHODS: A total of 60 calves on two farms were followed from the 2nd week of life through the 11th week of life and assessed on a weekly basis for CRS and lung consolidation via TUS. The alignment of BRD diagnoses based on CRSC scores and TUS findings was evaluated across disease progression (pre-consolidation, onset, chronic, or recovered) and severity (lobular or lobar lung consolidation) using receiver operator curves and area under the curves combined with Cohen's kappa (κ), sensitivity, and specificity. RESULTS: The diagnosis of BRD using CRSC scores ≥5 aligned best with the onset of lobar lung consolidation (>1 cm in width and full thickness). This equated to an acceptable level of discrimination (AUC = 0.76), fair agreement (κ = 0.37), and a sensitivity of 29% and specificity of 99%. Similarly, there was acceptable discrimination (AUC = 0.70) and fair agreement (κ = 0.33) between CRSC ≥5 and the onset of a less severe threshold of disease based on lobular (1-3 cm2 but not full thickness) or lobar consolidation. Discrimination remained acceptable (AUC = 0.71) with fair agreement (κ = 0.28) between CRSC scores ≥2 for nasal discharge and/or cough (spontaneous or induced) and the onset of lobar consolidation. However, sensitivity was <40% across comparisons and outside of the onset of disease there tended to be poor discrimination, slight agreement, and lowered sensitivity between CRS and TUS diagnoses of lobular or lobar consolidation (pre-consolidation, chronic, or recovered). Conversely, specificity was relatively high (≥92%) across comparisons suggesting that CRSC diagnoses indicative of BRD and associated lung consolidation tend to result in few false positive diagnoses and accurate identification of healthy animals. CONCLUSIONS AND CLINICAL RELEVANCE: Although we found the specificity of clinical signs for diagnosing lung consolidation to be ≥92% across all methods of TUS evaluations, the low levels of sensitivity dictate that clinical assessments lead to many false negative diagnoses. Consequently, depending on clinical signs alone to diagnose BRD within populations of dairy calves will likely result in overlooking a substantial proportion of subclinically affected animals that could inform the success of treatment and prevention protocols and guide management decisions.

Transcriptional changes detected in fecal RNA from neonatal dairy calves of different breeds following gastrointestinal disease of varying severity.

Gastrointestinal (GI) disease is a major health concern in preweaned dairy calves. The objective of this fixed cohort study was to use RNA isolated from preweaned Holstein and Jersey heifer calf feces to study the molecular adaptations to variable clinical GI disease. The study was conducted on a commercial calf ranch in the western U.S. Enrolled calves were assessed twice daily for variations in demeanor, milk intake, and hydration. Fecal consistency scores were recorded at enrollment (day 1), and on the day (day 10) that a fecal sample was collected for differential gene expression (DGE). Calves with diarrhea on either day were classified as having either uncomplicated, localized GI disease (scours), or systemic GI disease (systemic enteritis). Eighty-four calves' fecal RNA was evaluated for DGE, of which 33 calves (n = 20 Holstein; n = 13 Jersey) were consistently healthy. The remaining 51 calves (n = 23 Holstein; n = 28 Jersey) experienced varying severity of GI disease during the sampling window. Genes of interest were related to the inflammatory response (i.e., IFNG, NFKB1, NOD2, TLR2, and TLR4) and cell membrane or cytoplasmic transport (i.e., AQP3, FABP2, KRT8 and SLC5A1). Breed-specific findings indicated that AQP3, IFNG, and TLR4 were upregulated in Holsteins with systemic enteritis, whereas KRT8 was downregulated in systemically affected Jerseys. Holsteins did not appear affected by scours aside from a tendency for DGE of toll-like receptors (TLRs) on the day of diarrhea. However, Jersey calves consistently demonstrated a tendency to upregulate IFNG, NFKB1, and TLR4 when affected with either scours or systemic enteritis. These findings were more pronounced in systemically affected Jersey calves and were observed as a delayed response to both scours and systemic enteritis. These findings support previous observations suggesting that Holstein calves may be better equipped than Jersey calves to rapidly fight pathogen invasion.

Clarifying dairy calf mortality phenotypes through postmortem analysis.

Health problems can be thought of as phenotypic expressions of the complex relationships between genes, environments, and phenomes as a whole. Detailed evaluations of phenotypic expressions of illness are required to characterize important biological outcomes. We hypothesized that classifying dairy calf mortality phenotypes via a systematic postmortem analysis would identify different cause-of-death diagnoses than those derived from treatments alone. This cross-sectional study was carried out on a dairy calf ranch in the northwestern United States from June to September 2017 and focused on calves ≤90 d of age. Comparisons were made between causes of death based on 3 levels of information: on-farm treatment records alone, necropsy-based postmortem analyses in addition to treatment records, and Washington Animal Disease Diagnostic Laboratory (WADDL) results in addition to all other information. A total of 210 dairy calves were necropsied during this study, of which 122 cases were submitted to WADDL. Necropsy- and WADDL-derived mortality phenotypes were in almost perfect agreement (Cohen's κ = 0.86) when broadly categorized as diarrhea, respiratory, diarrhea and respiratory combined, or other causes. The level of agreement between on-farm treatment records and postmortem-derived results was low and varied by the level of diagnostic detail provided. There was just fair agreement (κ = 0.22) between treatment-based and necropsy-based phenotypes without WADDL input and only slight agreement (κ = 0.13) between treatment-based and corresponding necropsy-based phenotypes with WADDL input. Even for those cases in which causes of death aligned along a comparable pathologic spectrum, the lack of detail inherent to standard treatment-based causes of death failed to identify meaningful target areas for intervention. This was especially apparent for numerous cases of necrotizing enteritis and typhlitis (cecal inflammation) that were variously categorized as diarrhea and pneumonia by treatment-based diagnoses. The specificity of these lesions stood in stark contrast to the otherwise generic cause of death diagnoses derived from treatments. The findings from this study supported the hypothesis and highlighted the value of on-farm necropsies and laboratory-based diagnostics to (1) detect antemortem disease misclassifications, (2) provide detail regarding disease processes and mortality phenotypes, and (3) direct disease mitigation strategies.

Preweaned heifer management on US dairy operations: Part IV. Factors associated with the presence of Escherichia coli O157 in preweaned dairy heifers.

Dairy calves shed pathogenic Escherichia coli O157 (O157) in feces and are a potential route of exposure for human infections. As part of the National Animal Health Monitoring System's (NAHMS) Dairy 2014 study, we evaluated farm, animal, and environmental factors associated with O157 presence in dairy heifer calves. For this O157 study, calves were enrolled from 100 dairy operations in 13 states. Each operation collected data from calves from birth to weaning over an 18-mo period. A single fecal sample was collected from 487 calves in western states and from 871 calves in eastern states (n = 1,358 total), and O157 was detected in 2.5% (n = 34) of fecal samples. Descriptive statistics and univariable screening were used to determine which farm practices, environmental factors, and calf health measures were associated with O157 detection. Multilevel logistic models, controlling for dairy operation, were created using backward elimination of screened variables. The final O157 main effects model included variables for source of colostrum, temperature-humidity index (THI), and serum IgG concentration. Higher serum IgG was associated with lower odds of O157 shedding, whereas calves fed colostrum from their own dam had higher odds of O157 shedding than calves fed colostrum from pooled sources. Interaction models showed that THI level modified the effect of colostrum source on O157 shedding; calves with a THI indicative of heat stress had a significantly increased presence of O157 when fed colostrum from a first-lactation dam. The THI level also modified the effects of serum IgG. Calves with thermoneutral or heat stress THI values had increased presence of O157 with poor (<10 g/L) or adequate (10-15 g/L) serum IgG levels compared with those having excellent (≥15 g/L) serum IgG levels. These results highlight factors that influence the presence of O157 in preweaned dairy heifer calves and may be used to guide practices that mitigate shedding through improved animal husbandry.

Evaluation of horn bud wound healing following cautery disbudding of preweaned dairy calves treated with aluminum-based aerosol bandage.

Pain management during and following disbudding procedures has been studied extensively, though few studies have evaluated wound healing following cautery disbudding in dairy calves. The purpose of this study was to observe wound healing following cautery disbudding with or without treatment using a topical aluminum-based aerosol bandage (ALU) in preweaned dairy calves. Dairy calves were disbudded within the first 3 wk of life using a standard cautery disbudding protocol. The ALU treatment was randomly allocated to the right or left horn bud within each animal. The outcomes measured were lesion score (LS) and wound diameter (WD). The LS was evaluated on a scale of 1 to 3, with LS = 1 representing normal healing without a scab or exudate, LS = 2 having the presence of a scab, and LS = 3 showing the presence of wound exudate. Lesion score and WD were evaluated on a weekly basis following dehorning for 3 wk. A total of 209 animals completed the study. No difference was observed in LS between groups during the first 2 wk postdisbudding, but the proportion of LS = 3 on wk 3 postdisbudding was greater for the control group when compared with ALU (17 vs. 8%, respectively). During wk 1 and 2 postdisbudding, the odds of having delayed healing, or a LS ≥2, were similar for both groups. However, the odds tended to be different at wk 3 postdisbudding with control disbudding sites being 1.42 times more likely to have delayed healing than ALU. In wk 3, WD was 1 mm smaller in the treatment group compared with the control, and treatment decreased diameter over time compared with controls. Overall, once abnormal wound healing was observed, the likelihood of having abnormal wound healing the following week was increased. However, treatment with ALU diminished this effect on delayed healing during the follow-up period. Based on these results, the use of ALU improved wound healing following cautery disbudding of preweaned dairy calves.

Daily shedding dynamics of E. coli O157 in an Australian grass-fed beef herd.

This study aimed to describe the diurnal shedding dynamics of Escherichia coli O157 in cattle managed on pasture. The purpose was to identify the value of a single measurement for predicting the shedding status on subsequent days. Over a 14-day period, 24 beef cows with known E. coli O157 shedding status were sampled twice daily or daily (21 sampling points) and E. coli O157 was enumerated from faeces. No association between shedding status of individual animals within a 7-h period was identified (odds ratio 1·5, P = 0·08). Short-interval sampling demonstrated substantial diurnal volatility in shedding of E. coli O157 that is not evident in studies based on long-interval (>7 days) sampling. The findings contribute to and support previous findings on the question why it has been difficult to achieve progress in understanding the epidemiology of E. coli O157 infection in cattle.

Synchronization of E. coli O157 shedding in a grass-fed beef herd: a longitudinal study.

This study aims to describe in detail the temporal dynamics of E. coli O157 shedding and risk factors for shedding in a grass-fed beef herd. During a 9-month period, 23 beef cows were sampled twice a week (58 sampling points) and E. coli O157 was enumerated from faecal samples. Isolates were screened by PCR for presence of rfbE, stx 1 and stx 2 . The prevalence per sampling day ranged from 0% to 57%. This study demonstrates that many members of the herd were concurrently shedding E. coli O157. Occurrence of rainfall (P < 0·01), feeding silage (P < 0·01) and lactating (P < 0·01) were found to be predictors of shedding. Moving cattle to a new paddock had a negative effect on shedding. This approach, based on short-interval sampling, confirms the known variability of shedding within a herd and highlights that high shedding events are rare.

Conceptual modeling of postmortem evaluation findings to describe dairy cow deaths.

Dairy cow mortality levels in the United States are excessive and increasing over time. To better define cause and effect and combat rising mortality, clearer definitions of the reasons that cows die need to be acquired through thorough necropsy-based postmortem evaluations. The current study focused on organizing information generated from postmortem evaluations into a monitoring system that is based on the fundamentals of conceptual modeling and that will potentially be translatable into on-farm relational databases. This observational study was conducted on 3 high-producing, commercial dairies in northern Colorado. Throughout the study period a thorough postmortem evaluation was performed by veterinarians on cows that died on each dairy. Postmortem data included necropsy findings, life-history features (e.g., birth date, lactation number, lactational and reproductive status), clinical history and treatments, and pertinent aspects of operational management that were subject to change and considered integral to the poor outcome. During this study, 174 postmortem evaluations were performed. Postmortem evaluation results were conceptually modeled to view each death within the context of the web of factors influencing the dairy and the cow. Categories were formulated describing mortality in terms of functional characteristics potentially amenable to easy performance evaluation, management oversight, and research. In total, 21 death categories with 7 category themes were created. Themes included specific disease processes with variable etiologies, failure of disease recognition or treatment, traumatic events, multifactorial failures linked to transition or negative energy balance issues, problems with feed management, miscellaneous events not amenable to prevention or treatment, and undetermined causes. Although postmortem evaluations provide the relevant information necessary for framing a cow's death, a restructuring of on-farm databases is needed to integrate this level of detail into useful monitoring systems. Individual operations can focus on combating mortality through the use of employee training related to postmortem evaluations, detailed forms for capturing necropsy particulars and other relevant information related to deaths, and standardized nomenclature and categorization schemes. As much as anything, the simple act of recognizing mortality as a problem might be the most fundamental step toward controlling its progression.

Environmental mastitis in intensive high-producing dairy herds in New South Wales.

OBJECTIVE: To determine the prevalence of mastitis pathogens in high-producing intensive dairy herds in New South Wales. DESIGN: Field survey. PROCEDURE: Milk samples from the mastitis-affected quarter were collected from cows on five high-producing dairy farms in NSW. The 820 samples were cultured using standard microbiological culture techniques. RESULTS: Bacteria or fungi were isolated from 83.3% of samples (683/820). More than two colony types were isolated from 16.7% of samples (137/820), two types from 6.6% (54/820), and one type from 52.3% (429/820). No bacteria were isolated from 24.4% (200/820) of the primary cultures, but enrichment cultures of these samples yielded single colony type bacterial isolates from 36.5% (73/200) of samples. Environmental pathogens, including coliforms, environmental Streptococcus and Staphylococcus spp., made up 91% (555/610) of isolates and accounted for 33.6% (205/610), 41.6% (254/610) and 15.7% (96/610), respectively, of isolates. Escherichia coli accounted for 76.1% (156/205) of the coliform isolates, Streptococcus uberis and Streptococcus dysgalactiae accounted for 32.3% (82/254) and 28.0% (71/254), respectively, of the environmental streptococcal isolates. Contagious pathogens were uncommon, comprising only 2.5% (15/610) of the total isolates. CONCLUSION: The incidence and causes of mastitis are largely influenced by farm management. The relatively high prevalence of coliform mastitis in the intensive high-producing herds in this survey contrasts with the low incidence reported in surveys of pasture-based herds in Victoria. If the Australian dairy industry continues its current trend of intensification, coliform intra-mammary infections may emerge as an increasingly important cause of mastitis.

A necropsy-based descriptive study of dairy cow deaths on a Colorado dairy.

Increasing levels of dairy cow mortality pose a challenge to the US dairy industry. The industry's current understanding of dairy cow mortality is reliant upon descriptions largely based on producer or veterinary assumptions regarding cause of death without the benefit of detailed postmortem evaluations. A thorough necropsy is a superior tool for establishing a cause of death, except for cases involving euthanasia for traumatic accidents or severe locomotor disorders. Information provided from a necropsy examination would be most valuable if it were categorized and combined with cow health information in a complete postmortem evaluation designed to guide future management decisions. The objective of this study was to describe dairy cow deaths on a Colorado dairy over a 1-yr period and explore classification systems for necropsy findings that might inform management actions aimed at reducing dairy cow mortality. Throughout the study period a thorough necropsy examination was performed on every cow that died. Based upon this examination each death was characterized by a proximate cause (i.e., the most likely immediate cause of the death). Each proximate cause of death was then categorized using 3 alternate schemes founded on generalized etiologic principles and influenced by previous clinical history and treatments. These schemes included the broad categories commonly used for classifying findings within a review of literature related to dairy cow mortality, a diagnostic scheme used within the problem-oriented veterinary medical record, and an analysis focusing on the primary physiologic system derangement for each death. A total of 2,067 cows were enrolled during the study period of which 1,468 cows freshened, 507 cows were sold, and 94 cows died, resulting in a mortality risk of 6.4 deaths per 100 lactations at risk. The distribution of deaths by parity was significantly different from the herd distribution at the end of study with the largest percentage of death present in parity > or =4. Postmortem findings attributable to a specific cause of death were present for all but 4 of the 94 deaths. Assignment of the proximate causes of death to categories within the 3 alternate schemes provided a means for classifying necropsy findings and causes of death with different levels of detail. Creating categories with more selective groupings may provide a means for capturing specifics related to deaths that can be used to guide management decisions.

Evaluation of factors associated with increased dairy cow mortality on United States dairy operations.

Dairy cow mortality is an increasingly severe problem for the US dairy industry. The objective of this study was to examine a variety of herd management practices and herd characteristics to identify factors associated with increased cow mortality in US dairy herds. The National Animal Health Monitoring System's Dairy 2002 study surveyed dairy operations in 21 major dairy states. The complete data set included results from 953 dairy farms with a minimum of 30 dairy cows per farm. Associations between dairy cow mortality and 119 a priori-selected management practices or characteristics of 953 operations were evaluated. Eighty of the 119 risk factors explored in a univariate analysis met initial inclusion criteria for further evaluation of association with dairy cow mortality. A multivariable analysis was conducted to explore more complex relationships. The final multivariable model included 7 representative variables: herd levels of respiratory disease, lameness, and antibiotic use for treating sick cows, the percentage of culled cows less than 50 d in milk, the average calving interval, the use of a total mixed ration, and the region of the country. Increased odds of a greater level of mortality on farms was associated with greater percentages of lameness, respiratory disease, and sick cows treated with antibiotics, demonstrating the influence of physical derangements and disease on dairy cow mortality. Increased odds of a greater level of mortality was also associated with feeding a total mixed ration, culling fewer cows in early lactation, and herds located in western, midwestern, and southeastern regions relative to the northeastern United States, pointing to the importance of management decisions and operation characteristics on mortality outcomes. Further, an important interplay between facets of health and management on dairy cow mortality was suggested through the inclusion of the calving interval, with a longer calving interval leading to increased odds of a greater level of mortality on farms. Analysis of a variety of herd characteristics and practices with nationally representative data suggests that several health problems in tandem with aspects of operational construct and management are associated with increasing mortality.

Serologic cross-reactivity of Australian Moraxella bovis to vaccinal bacterin strains as determined by competitive ELISA.

OBJECTIVE: To conduct a serologic survey and define pili antigenic variability via the serologic cross-reactivity of Moraxella bovis isolates from naturally occurring infectious bovine keratoconjunctivitis (IBK) outbreaks in Australia. This project applies to the development of an M bovis pili-based vaccine targeting Australian strains originating from intensive cattle producing regions. PROCEDURE: Ocular swabs were collected from cattle affected with clinical signs of IBK from 25 veterinary practices. Standard criteria were used to identify 70 M bovis. Pure, piliated isolates were evaluated with a modified competitive enzyme-linked immunosorbent assay (ELISA) for cell-bound M bovis pili to determine their serologic cross-reactivity with pili of vaccinal bacterin strains EPP63, FLA64, and SAH 38. RESULTS: Sixty-four percent (45/70) of M bovis isolates demonstrated homologous pili antigens to a vaccinal strain. M bovis isolates homologous to one of the three vaccinal strains were obtained in 77% (34/44) of IBK outbreaks sampled. No IBK outbreak had isolates homologous to more than one vaccinal strain; however, 29% (10/34) of outbreaks with a cross-reacting strain had non-cross-reacting strains as well. CONCLUSION: The similar prevalence of pilus antigen homology to strain FLA64 was observed with isolates derived from NSW, Tasmania, and Victoria, compared with results of prior smaller serologic studies, suggests that the common pilus antigens in M bovis within Australia have been relatively stable over the last 20 years. The prevalence of a limited number of pilus antigens in M bovis suggest that the application of a vaccine containing the bacterial strains EPP63, FLA64, and SAH38 may provide a useful management tool for reducing production losses associated with IBK in Australia.

Infectious bovine keratoconjunctivitis antimicrobial therapy.

Infectious bovine keratoconjunctivitis (IBK) is one of the most common diseases of cattle and is of major economic importance. If the primary aetiological agent, Moraxella bovis, is successfully eliminated from ocular tissues corneal ulcers heal at a constant rate. If treatment is unsuccessful ulcer reoccurrence may follow initial healing. Appropriate antimicrobial selection requires knowledge of antimicrobial sensitivities and distribution in ocular tissues and tears. Drugs may be delivered to the eye in several ways: subconjunctival injection, topical application and systemic administration. While therapeutic efficacy is affected by the frequency and mode of drug delivery, variations between intensive and extensive enterprises dictate the practical method of antimicrobial delivery. Specific recommendations for antimicrobial therapies targeting Australian IBK outbreaks are dependent upon antimicrobial pharmacokinetics, drug regulations and associated costs.

Infectious bovine keratoconjunctivitis vaccine development.

Infectious bovine keratoconjunctivitis is a common and highly contagious ocular disease affecting cattle worldwide. The tremendous economic losses attributable to this disease warrant continued investigation into methods of prevention. Multiple virulence factors have been linked to the primary aetiologic agent, Moraxella bovis. Efforts to develop an efficacious vaccine have primarily focused upon the use of surface pili or cytolysin to stimulate host immunity; however, M. bovis possesses other virulence determinants that include proteases, fibrinolysins, phospholipases and other cell surface components such as outer membrane proteins. These potentially conserved antigens provide additional possibilities for vaccine development. Examination of appropriate antigen presentation is necessary to attain an adequate immune response. Further, the potential for antigenic diversity as well as epitope conversion requires continuous epidemiological surveillance of isolates recovered from outbreaks. Current work targeting conserved immunogens provides hope for efficacious vaccines that when used in tandem with proper management may control, if not prevent, infectious bovine keratoconjunctivitis.

Otologic evaluation of a patient with a suspected lesion of the central auditory system.

Patients with a lesion of the central auditory system often present with vague and ill-defined symptoms that require a high degree of suspicion on the part of the physician to institute the proper steps in diagnosis. This article establishes guidelines that can be followed by the otolaryngologist using equipment that is readily accessible in most community settings.