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1.
Vet Sci ; 10(1)2022 Dec 29.
Article in English | MEDLINE | ID: mdl-36669022

ABSTRACT

Maternal antibodies interfere with BRSV vaccine responses and efficacy in young calves. The objective of this study was to determine if vaccination before the complete absorption of colostral antibodies results in adequate immune priming and clinical protection of beef calves. Within 6 h of life, calves were randomly assigned to 2 different treatment groups. Group Vacc (n = 25) received a single dose of a modified-live virus (MLV) BRSV vaccine intranasally (IN) and group Control (n = 25) received 2 mL of 0.9% saline IN. At approximately 3 months of age, all calves were experimentally challenged with BRSV. Serum and nasal secretion samples were collected before and after challenge for BRSV real-time RT-PCR and antibody testing. Respiratory signs were not observed before challenge. After challenge, respiratory scores were similar between groups. On the challenge day, >40% of calves in each group were febrile. The mean serum and nasal BRSV-specific antibody titers indicated natural BRSV exposure before the experimental challenge in both groups. All calves tested positive for BRSV and had a similar duration of shedding after challenge. Based on these results, vaccination at birth does not offer advantages for immune priming or clinical protection for beef calves in BRSV-endemic cow-calf herds.

2.
Nat Mater ; 2022 Nov 28.
Article in English | MEDLINE | ID: mdl-36443576

ABSTRACT

Messenger RNA has now been used to vaccinate millions of people. However, the diversity of pulmonary pathologies, including infections, genetic disorders, asthma and others, reveals the lung as an important organ to directly target for future RNA therapeutics and preventatives. Here we report the screening of 166 polymeric nanoparticle formulations for functional delivery to the lungs, obtained from a combinatorial synthesis approach combined with a low-dead-volume nose-only inhalation system for mice. We identify P76, a poly-ß-amino-thio-ester polymer, that exhibits increased expression over formulations lacking the thiol component, delivery to different animal species with varying RNA cargos and low toxicity. P76 allows for dose sparing when delivering an mRNA-expressed Cas13a-mediated treatment in a SARS-CoV-2 challenge model, resulting in similar efficacy to a 20-fold higher dose of a neutralizing antibody. Overall, the combinatorial synthesis approach allowed for the discovery of promising polymeric formulations for future RNA pharmaceutical development for the lungs.

3.
Vet Clin Pathol ; 2022 Nov 06.
Article in English | MEDLINE | ID: mdl-36336840

ABSTRACT

An approximately 12-year-old female Vietnamese Pot-Bellied Pig was presented to the Mississippi State College of Veterinary Medicine Food Animal Service for anorexia of 2 days duration. On physical examination, the patient appeared depressed and lethargic with significantly pale mucus membranes, open mouth breathing, and nostril flaring. On abdominal palpation, the abdomen was tense and uncomfortable. A complete blood count (CBC) and chemistry profile were performed. The CBC revealed significant anemia and mild leukocytosis characterized by mild neutrophilia with a left shift. Mast cells were rarely observed. Hematocrit = 8.1% (RI 22-50), RBC = 1.25 × 106 /µL (RI 3.6-7.8), WBC = 19.85 × 103 /µL (RI 5.2-17.9), Neutrophils = 15.08 × 103 /µL (RI 0-11.4), and Bands = 0.993 × 103 /µL (RI 0-0.019). The chemistry profile was unremarkable with a mildly elevated BUN and slightly decreased total protein and albumin (BUN = 39 mg/dL [RI 4.2-15.1], total protein = 6.2 g/dL [RI 6.6-8.9], and albumin = 2.5 g/dL [RI 3.6-5.0]). An abdominal ultrasound revealed numerous hypoechoic nodules diffusely scattered throughout the hepatic parenchyma. An FNA of one of the hepatic nodules was performed. A mild suppurative component and numerous variably granulated mast cells were observed. A presumptive cytologic diagnosis of mast cell tumor was made. Histopathology was performed, confirming the cytologic interpretation.

4.
PLoS One ; 17(11): e0277033, 2022.
Article in English | MEDLINE | ID: mdl-36327246

ABSTRACT

Bovine respiratory disease (BRD), the leading disease complex in beef cattle production systems, remains highly elusive regarding diagnostics and disease prediction. Previous research has employed cellular and molecular techniques to describe hematological and gene expression variation that coincides with BRD development. Here, we utilized weighted gene co-expression network analysis (WGCNA) to leverage total gene expression patterns from cattle at arrival and generate hematological and clinical trait associations to describe mechanisms that may predict BRD development. Gene expression counts of previously published RNA-Seq data from 23 cattle (2017; n = 11 Healthy, n = 12 BRD) were used to construct gene co-expression modules and correlation patterns with complete blood count (CBC) and clinical datasets. Modules were further evaluated for cross-populational preservation of expression with RNA-Seq data from 24 cattle in an independent population (2019; n = 12 Healthy, n = 12 BRD). Genes within well-preserved modules were subject to functional enrichment analysis for significant Gene Ontology terms and pathways. Genes which possessed high module membership and association with BRD development, regardless of module preservation ("hub genes"), were utilized for protein-protein physical interaction network and clustering analyses. Five well-preserved modules of co-expressed genes were identified. One module ("steelblue"), involved in alpha-beta T-cell complexes and Th2-type immunity, possessed significant correlation with increased erythrocytes, platelets, and BRD development. One module ("purple"), involved in mitochondrial metabolism and rRNA maturation, possessed significant correlation with increased eosinophils, fecal egg count per gram, and weight gain over time. Fifty-two interacting hub genes, stratified into 11 clusters, may possess transient function involved in BRD development not previously described in literature. This study identifies co-expressed genes and coordinated mechanisms associated with BRD, which necessitates further investigation in BRD-prediction research.


Subject(s)
Bovine Respiratory Disease Complex , Cattle Diseases , Respiration Disorders , Respiratory Tract Diseases , Cattle , Animals , Respiratory Tract Diseases/genetics , Respiratory System , Gene Regulatory Networks , Weight Gain/genetics , Bovine Respiratory Disease Complex/genetics
5.
Front Vet Sci ; 9: 1010039, 2022.
Article in English | MEDLINE | ID: mdl-36225796

ABSTRACT

The impact of preweaning vaccination for bovine respiratory viruses on cattle health and subsequent bovine respiratory disease morbidity has been widely studied yet questions remain regarding the impact of these vaccines on host response and gene expression. Six randomly selected calves were vaccinated twice preweaning (T1 and T3) with a modified live vaccine for respiratory pathogens and 6 randomly selected calves were left unvaccinated. Whole blood samples were taken at first vaccination (T1), seven days later (T2), at revaccination and castration (T3), and at weaning (T4), and utilized for RNA isolation and sequencing. Serum from T3 and T4 was analyzed for antibodies to BRSV, BVDV1a, and BHV1. Sequenced RNA for all 48 samples was bioinformatically processed with a HISAT2/StringTie pipeline, utilizing reference guided assembly with the ARS-UCD1.2 bovine genome. Differentially expressed genes were identified through analyzing the impact of time across all calves, influence of vaccination across treatment groups at each timepoint, and the interaction of time and vaccination. Calves, regardless of vaccine administration, demonstrated an increase in gene expression over time related to specialized proresolving mediator production, lipid metabolism, and stimulation of immunoregulatory T-cells. Vaccination was associated with gene expression related to natural killer cell activity and helper T-cell differentiation, enriching for an upregulation in Th17-related gene expression, and downregulated genes involved in complement system activity and coagulation mechanisms. Type-1 interferon production was unaffected by the influence of vaccination nor time. To our knowledge, this is the first study to evaluate mechanisms of vaccination and development in healthy calves through RNA sequencing analysis.

6.
Am J Vet Res ; 83(11)2022 Oct 04.
Article in English | MEDLINE | ID: mdl-36173761

ABSTRACT

OBJECTIVES: To compare initial titers, duration, and residual clinical protection of passively transferred bovine respiratory syncytial virus (BRSV) nasal immunoglobulin (Ig) G-1 and IgA, and serum neutralizing (SN) antibodies. ANIMALS: 40 three-month-old beef steers born either to unvaccinated or vaccinated cows. PROCEDURES: During the last trimester of gestation, cows were assigned randomly to either vaccinated or unvaccinated groups. Calves were grouped on the basis of whether they nursed colostrum from unvaccinated dams (NO-VACC group; n = 20) versus dams vaccinated with 2 doses of an inactivated BRSV vaccine (VACC group; n = 20). At 3 months of age, calves were challenged with BRSV. Respiratory signs were scored. Nasal BRSV IgG-1 and IgA and SN antibodies were compared before and after the challenge. The presence of BRSV in nasal secretions was evaluated by reverse transcription-PCR assays. RESULTS: Respiratory scores after BRSV challenge were similar between treatment groups. Nasal BRSV IgG-1 and SN antibodies were significantly greater in VACC calves at 48 hours of life; however, by 3 months of age, titers had decayed in both groups. Nasal BRSV IgA titers were minimal after colostrum intake and before the BRSV challenge, and increased in both groups after the challenge. The NO-VACC group had a significantly greater probability of shedding BRSV compared with VACC calves. CLINICAL RELEVANCE: At 3 months of age, titers of passively transferred BRSV antibodies in VACC and NO-VACC calves had decayed to nonprotective levels. Calves born to vaccinated dams had a decreased probability of BRSV shedding; however, this was not related to differences in SN or nasal BRSV antibody titers.


Subject(s)
Cattle Diseases , Respiratory Syncytial Virus Infections , Respiratory Syncytial Virus, Bovine , Pregnancy , Female , Cattle , Animals , Colostrum , Cattle Diseases/prevention & control , Antibodies, Viral , Immunoglobulin G , Immunoglobulin A , Respiratory Syncytial Virus Infections/prevention & control , Respiratory Syncytial Virus Infections/veterinary
7.
Anim Microbiome ; 4(1): 49, 2022 Aug 13.
Article in English | MEDLINE | ID: mdl-35964128

ABSTRACT

BACKGROUND: Bovine respiratory disease (BRD) is caused by interactions among host, environment, and pathogens. One standard method for antemortem pathogen identification in cattle with BRD is deep-guarded nasopharyngeal swabbing, which is challenging, costly, and waste generating. The objective was to compare the ability to recover Mannheimia haemolytica and compare microbial community structure using 29.5 inch (74.9 cm) deep-guarded nasopharyngeal swabs, 16 inch (40.6 cm) unguarded proctology swabs, or 6 inch (15.2 cm) unguarded nasal swabs when characterized using culture, real time-qPCR, and 16S rRNA gene sequencing. Samples for aerobic culture, qPCR, and 16S rRNA gene sequencing were collected from the upper respiratory tract of cattle 2 weeks after feedlot arrival. RESULTS: There was high concordance of culture and qPCR results for all swab types (results for 77% and 81% of sampled animals completely across all 3 swab types for culture and qPCR respectively). Microbial communities were highly similar among samples collected with different swab types, and differences identified relative to treatment for BRD were also similar. Positive qPCR results for M. haemolytica were highly concordant (81% agreed completely), but samples collected by deep-guarded swabbing had lower amounts of Mh DNA identified (Kruskal-Wallis analysis of variance on ranks, P < 0.05; Dunn-test for pairwise comparison with Benjamini-Hochberg correction, P < 0.05) and lower frequency of positive compared to nasal and proctology swabs (McNemar's Chi-square test, P < 0.05). CONCLUSIONS: Though differences existed among different types of swabs collected from individual cattle, nasal swabs and proctology swabs offer comparable results to deep-guarded nasopharyngeal swabs when identifying and characterizing M. haemolytica by culture, 16S rRNA gene sequencing, and qPCR.

8.
J Dairy Sci ; 105(9): 7750-7763, 2022 Sep.
Article in English | MEDLINE | ID: mdl-35931482

ABSTRACT

Bovine respiratory disease (BRD) is a multifactorial disease which causes short- and long-term negative effects. Early detection is crucial for a prompt response to therapy, as well as to decrease mortality risk. Clinical scoring systems have been developed mostly in North America for screening calves at risk or suspected of having BRD, and these tools have also been applied in subtropical and tropical countries. However, it has been unknown whether these scoring systems had the same accuracy in tropical environmental conditions. Therefore, this study evaluated the accuracy of 4 different field techniques, as well as serum haptoglobin (HAP), to diagnose BRD in Holstein dairy calves in subtropical conditions. The tests used to diagnose BRD were thoracic ultrasound (TUS; positive if consolidation depth ≥1 cm), thoracic auscultation (AUSC; positive if crackles, wheezes, or silent areas were present), Wisconsin score (WISC; ≥2 categories with scores of ≥2), and California score (CALIF; positive if total score ≥5). Also, HAP was measured and classified as positive if ≥15 mg/dL. Heifers between 30 d of age and weaning (n = 482), residing on 17 commercial dairies in São Paulo state, were enrolled in this study. Bayesian latent class models were used with informative priors to evaluate the accuracy of TUS, AUSC, and HAP, and noninformative priors for the accuracy of WISC and CALIF. The percentage of calves positive for each test on each farm ranged from 0 to 56% for WISC, 11-51% for CALIF, 0-72% for TUS, 0-32% for AUSC, and 0-100% for HAP. The sensitivity (Se; 95% credible interval) and specificity (Sp) for WISC were 77.9% (64.8-90.2) and 81.9% (76.3-88.2). For CALIF, the Se was 67.1% (53.6-80.1) and Sp 79.1% (73.9-84.6). For TUS Se was 59.8% (46.5-73.1) and Sp was 84.8% (80.0-89.5), and for AUSC, Se was 58.8% (41.3-79.8) and Sp was 98.6% (95.7-99.9). The Se and Sp of HAP was 67.6% (55.3-78.8) and 46.7% (41.4-52.2), respectively. The performance of the scoring systems was similar to, or better than, the performance found in North American studies, despite the fact that calves were in a tropical environment.


Subject(s)
Bovine Respiratory Disease Complex , Cattle Diseases , Respiratory Tract Diseases , Animals , Bayes Theorem , Bovine Respiratory Disease Complex/diagnosis , Brazil , California , Cattle , Female , Respiratory Tract Diseases/veterinary , Wisconsin
9.
J Anim Sci ; 100(11)2022 Nov 01.
Article in English | MEDLINE | ID: mdl-35926833

ABSTRACT

Experimental bovine respiratory syncytial virus (BRSV) infection can enhance Histophilus somni (Hs) disease in calves; we thus hypothesized that modified-live virus (MLV) vaccines containing BRSV may alter Hs carriage. Our objective was to determine the effects of an intranasal (IN) trivalent (infectious bovine rhinotracheitis virus [IBRV], parainfluenza-3 virus [PI3V], and BRSV) respiratory vaccine with parenteral (PT) bivalent bovine viral diarrhea virus (BVDV) type I + II vaccine, or a PT pentavalent (BVDV type I and II, IBRV, BRSV, and PI3V) respiratory vaccine, on health, growth, immunity, and nasal pathogen colonization in high-risk beef calves. Calves (n = 525) were received in five truckload blocks and stratified by body weight (213 ± 18.4 kg), sex, and presence of a pre-existing ear-tag. Pens were spatially arranged in sets of three within a block and randomly assigned to treatment with an empty pen between treatment groups consisting of: 1) no MLV respiratory vaccination (CON), 2) IN trivalent MLV respiratory vaccine with PT BVDV type I + II vaccine (INT), or 3) PT pentavalent, MLV respiratory vaccine (INJ). The pen was the experimental unit, with 15 pens/treatment and 11 to 12 calves/pen in this 70-d receiving study. Health, performance, and BRSV, Hs, Mycoplasma bovis (Mb), Mannheimia haemolytica (Mh), and Pasteurella multocida (Pm) level in nasal swabs via rtPCR was determined on days 0, 7, 14, and 28, and BRSV-specific serum neutralizing antibody titer, and serum IFN-γ concentration via ELISA, were evaluated on days 0, 14, 28, 42, 56, and 70. Morbidity (P = 0.83), mortality (P = 0.68) and average daily gain (P ≥ 0.82) did not differ. Serum antibodies against BRSV increased with time (P < 0.01). There was a treatment × time interaction (P < 0.01) for Hs detection; on days 14 and 28, INT (21.1% and 57.1%) were more frequently (P < 0.01) Hs positive than CON (3.6% and 25.3%) or INJ (3.4 % and 8.4%). Also, INT had reduced (P = 0.03) cycle time of Hs positive samples on day 28. No difference (P ≥ 0.17) was found for IFN-γ concentration and Mb, Mh, or Pm detection. The proportion of Mh positive culture from lung specimens differed (P < 0.01); INT had fewer (0.0%; 0 of 9) Mh positive lungs than INJ (45.5%; 6 of 13) or CON (74.0%; 14 of 19). Vaccination of high-risk calves with MLV did not clearly impact health or growth during the receiving period. However, INT was associated with an altered upper respiratory microbial community in cattle resulting in increased detection and level of Hs.


Our objective was to determine the safety, efficiency, and effects on immunity and nasal shedding of respiratory pathogens for high-risk cattle administered an intranasal (IN), trivalent (infectious bovine rhinotracheitis virus [IBRV], parainfluenza-3 virus [PI3V], and bovine respiratory syncytial virus [BRSV]) respiratory vaccine with parenteral, bivalent bovine viral diarrhea virus (BVDV), or a parenteral, pentavalent (BVDV type I and II, IBRV, BRSV, and PI3V) respiratory vaccine, compared to an unvaccinated negative control. The results of this study indicate that modified-live virus (MLV) vaccination of high-risk calves upon arrival, either parenterally or intranasally, did not clearly impact health or growth during the feedlot receiving period. However, cattle that were intranasally vaccinated had increased carriage of Histophilus somni in the naris, greater amount of H. somni in nasal swabs indicated by reduced PCR cycle time, and less frequent culture of Mannheimia haemolytica from lung tissue samples upon necropsy. Therefore, intranasal administration of MLV vaccines may alter the microbial community and balance of opportunistic pathogens in the respiratory tract of cattle.


Subject(s)
Cattle Diseases , Diarrhea Viruses, Bovine Viral , Herpesvirus 1, Bovine , Mannheimia haemolytica , Pasteurella multocida , Respiratory Syncytial Virus, Bovine , Viral Vaccines , Cattle , Animals , Antibodies, Viral , Vaccines, Attenuated , Cattle Diseases/prevention & control
10.
Vet Clin North Am Food Anim Pract ; 38(2): 219-227, 2022 Jul.
Article in English | MEDLINE | ID: mdl-35691625

ABSTRACT

Throughout history, theories of disease etiology have evolved. These theories of disease etiology, which can also be considered mental models of disease, have been based on associations drawn from careful observation of diseased and healthy individuals. Mental models of disease, even when incorrect, have frequently reflected real associations between proposed exposures and disease even when the exposures eventually were disproved as causal. The same patterns can be observed in mental models of disease in cattle. Throughout time, mental models for common bovine diseases have been improved to better reflect how disease actually occurs. It is important to recognize that inconsistencies still exist between observation of actual disease and our understanding of disease etiology. These inconsistencies can be viewed as opportunities for further discovery to improve our understanding of disease. Future progress in controlling bovine diseases depends on converting these opportunities into better mental models of disease.


Subject(s)
Cattle Diseases , Animals , Cattle , Models, Psychological
11.
Front Vet Sci ; 9: 883389, 2022.
Article in English | MEDLINE | ID: mdl-35647109

ABSTRACT

The threat of bovine respiratory disease (BRD) for cattle operations is exacerbated by increasing prevalence of antimicrobial resistance (AMR) in Mannheimia haemolytica, a leading cause of BRD. Characterization of AMR in M. haemolytica by culture and susceptibility testing is complicated by uncertainty regarding the number of colonies that must be selected to accurately characterize AMR phenotypes (antibiograms) and genotypes in a culture. The study objective was to assess phenotypic and genotypic diversity of M. haemolytica isolates on nasopharyngeal swabs (NPS) from 28 cattle at risk for BRD or with BRD. NPS were swabbed onto five consecutive blood agar plates; after incubation up to 20 M. haemolytica colonies were selected per plate (up to 100 colonies per NPS). Phenotype was determined by measuring minimum inhibitory concentrations (MIC) for 11 antimicrobials and classifying isolates as resistant or not. Genotype was indirectly determined by matrix-assisted laser desorption/ionization time of flight mass spectroscopy (MALDI-TOF MS). NPS from 11 of 28 cattle yielded at least one M. haemolytica isolate; median (range) of isolates per NPS was 48 (1-94). NPS from seven cattle yielded one phenotype, 3 NPS yielded two, and 1 NPS yielded three; however, within a sample all phenotypic differences were due to only one MIC dilution. On each NPS all M. haemolytica isolated were the same genotype; genotype 1 was isolated from three NPS and genotype two was isolated from eight. Diversity of M. haemolytica on bovine NPS was limited, suggesting that selection of few colonies might adequately identify relevant phenotypes and genotypes.

12.
BMC Vet Res ; 18(1): 77, 2022 Feb 23.
Article in English | MEDLINE | ID: mdl-35197051

ABSTRACT

BACKGROUND: Transcriptomics has identified at-arrival differentially expressed genes associated with bovine respiratory disease (BRD) development; however, their use as prediction molecules necessitates further evaluation. Therefore, we aimed to selectively analyze and corroborate at-arrival mRNA expression from multiple independent populations of beef cattle. In a nested case-control study, we evaluated the expression of 56 mRNA molecules from at-arrival blood samples of 234 cattle across seven populations via NanoString nCounter gene expression profiling. Analysis of mRNA was performed with nSolver Advanced Analysis software (p < 0.05), comparing cattle groups based on the diagnosis of clinical BRD within 28 days of facility arrival (n = 115 Healthy; n = 119 BRD); BRD was further stratified for severity based on frequency of treatment and/or mortality (Treated_1, n = 89; Treated_2+, n = 30). Gene expression homogeneity of variance, receiver operator characteristic (ROC) curve, and decision tree analyses were performed between severity cohorts. RESULTS: Increased expression of mRNAs involved in specialized pro-resolving mediator synthesis (ALOX15, HPGD), leukocyte differentiation (LOC100297044, GCSAML, KLF17), and antimicrobial peptide production (CATHL3, GZMB, LTF) were identified in Healthy cattle. BRD cattle possessed increased expression of CFB, and mRNA related to granulocytic processes (DSG1, LRG1, MCF2L) and type-I interferon activity (HERC6, IFI6, ISG15, MX1). Healthy and Treated_1 cattle were similar in terms of gene expression, while Treated_2+ cattle were the most distinct. ROC cutoffs were used to generate an at-arrival treatment decision tree, which classified 90% of Treated_2+ individuals. CONCLUSIONS: Increased expression of complement factor B, pro-inflammatory, and type I interferon-associated mRNA hallmark the at-arrival expression patterns of cattle that develop severe clinical BRD. Here, we corroborate at-arrival mRNA markers identified in previous transcriptome studies and generate a prediction model to be evaluated in future studies. Further research is necessary to evaluate these expression patterns in a prospective manner.


Subject(s)
Bovine Respiratory Disease Complex , Cattle Diseases , Animals , Bovine Respiratory Disease Complex/diagnosis , Bovine Respiratory Disease Complex/genetics , Case-Control Studies , Cattle , Cattle Diseases/diagnosis , Prospective Studies , RNA, Messenger/genetics , Transcriptome
13.
Molecules ; 27(2)2022 Jan 16.
Article in English | MEDLINE | ID: mdl-35056864

ABSTRACT

Bovine respiratory syncytial virus (BRSV) is a major contributor to respiratory disease in cattle worldwide. Traditionally, BRSV infection is detected based on non-specific clinical signs, followed by reverse transcriptase-polymerase chain reaction (RT-PCR), the results of which can take days to obtain. Near-infrared aquaphotomics evaluation based on biochemical information from biofluids has the potential to support the rapid identification of BRSV infection in the field. This study evaluated NIR spectra (n = 240) of exhaled breath condensate (EBC) from dairy calves (n = 5) undergoing a controlled infection with BRSV. Changes in the organization of the aqueous phase of EBC during the baseline (pre-infection) and infected (post-infection and clinically abnormal) stages were found in the WAMACS (water matrix coordinates) C1, C5, C9, and C11, likely associated with volatile and non-volatile compounds in EBC. The discrimination of these chemical profiles by PCA-LDA models differentiated samples collected during the baseline and infected stages with an accuracy, sensitivity, and specificity >93% in both the calibration and validation. Thus, biochemical changes occurring during BRSV infection can be detected and evaluated with NIR-aquaphotomics in EBC. These findings form the foundation for developing an innovative, non-invasive, and in-field diagnostic tool to identify BRSV infection in cattle.


Subject(s)
Breath Tests/methods , Cattle Diseases/diagnosis , Respiratory Syncytial Virus Infections/diagnosis , Spectroscopy, Near-Infrared/methods , Animals , Calibration , Cattle , Cattle Diseases/virology , Cell Line , Photometry/methods , Respiratory Syncytial Virus Infections/virology , Respiratory Syncytial Virus, Bovine/isolation & purification , Sensitivity and Specificity , Water/analysis , Water/chemistry
14.
Sci Rep ; 11(1): 23877, 2021 12 13.
Article in English | MEDLINE | ID: mdl-34903778

ABSTRACT

Bovine respiratory disease (BRD) remains the leading infectious disease in post-weaned beef cattle. The objective of this investigation was to contrast the at-arrival blood transcriptomes from cattle derived from two distinct populations that developed BRD in the 28 days following arrival versus cattle that did not. Forty-eight blood samples from two populations were selected for mRNA sequencing based on even distribution of development (n = 24) or lack of (n = 24) clinical BRD within 28 days following arrival; cattle which developed BRD were further stratified into BRD severity cohorts based on frequency of antimicrobial treatment: treated once (treated_1) or treated twice or more and/or died (treated_2+). Sequenced reads (~ 50 M/sample, 150 bp paired-end) were aligned to the ARS-UCD1.2 bovine genome assembly. One hundred and thirty-two unique differentially expressed genes (DEGs) were identified between groups stratified by disease severity (healthy, n = 24; treated_1, n = 13; treated_2+, n = 11) with edgeR (FDR ≤ 0.05). Differentially expressed genes in treated_1 relative to both healthy and treated_2+ were predicted to increase neutrophil activation, cellular cornification/keratinization, and antimicrobial peptide production. Differentially expressed genes in treated_2+ relative to both healthy and treated_1 were predicted to increase alternative complement activation, decrease leukocyte activity, and increase nitric oxide production. Receiver operating characteristic (ROC) curves generated from expression data for six DEGs identified in our current and previous studies (MARCO, CFB, MCF2L, ALOX15, LOC100335828 (aka CD200R1), and SLC18A2) demonstrated good-to-excellent (AUC: 0.800-0.899; ≥ 0.900) predictability for classifying disease occurrence and severity. This investigation identifies candidate biomarkers and functional mechanisms in at arrival blood that predicted development and severity of BRD.


Subject(s)
Cattle Diseases/genetics , Cattle/genetics , Respiratory Tract Infections/genetics , Transcriptome , Animals , Biomarkers/metabolism , Cattle/physiology , Respiratory Tract Infections/veterinary
15.
Sci Rep ; 11(1): 22916, 2021 11 25.
Article in English | MEDLINE | ID: mdl-34824337

ABSTRACT

Bovine respiratory disease (BRD) is a multifactorial disease involving complex host immune interactions shaped by pathogenic agents and environmental factors. Advancements in RNA sequencing and associated analytical methods are improving our understanding of host response related to BRD pathophysiology. Supervised machine learning (ML) approaches present one such method for analyzing new and previously published transcriptome data to identify novel disease-associated genes and mechanisms. Our objective was to apply ML models to lung and immunological tissue datasets acquired from previous clinical BRD experiments to identify genes that classify disease with high accuracy. Raw mRNA sequencing reads from 151 bovine datasets (n = 123 BRD, n = 28 control) were downloaded from NCBI-GEO. Quality filtered reads were assembled in a HISAT2/Stringtie2 pipeline. Raw gene counts for ML analysis were normalized, transformed, and analyzed with MLSeq, utilizing six ML models. Cross-validation parameters (fivefold, repeated 10 times) were applied to 70% of the compiled datasets for ML model training and parameter tuning; optimized ML models were tested with the remaining 30%. Downstream analysis of significant genes identified by the top ML models, based on classification accuracy for each etiological association, was performed within WebGestalt and Reactome (FDR ≤ 0.05). Nearest shrunken centroid and Poisson linear discriminant analysis with power transformation models identified 154 and 195 significant genes for IBR and BRSV, respectively; from these genes, the two ML models discriminated IBR and BRSV with 100% accuracy compared to sham controls. Significant genes classified by the top ML models in IBR (154) and BRSV (195), but not BVDV (74), were related to type I interferon production and IL-8 secretion, specifically in lymphoid tissue and not homogenized lung tissue. Genes identified in Mannheimia haemolytica infections (97) were involved in activating classical and alternative pathways of complement. Novel findings, including expression of genes related to reduced mitochondrial oxygenation and ATP synthesis in consolidated lung tissue, were discovered. Genes identified in each analysis represent distinct genomic events relevant to understanding and predicting clinical BRD. Our analysis demonstrates the utility of ML with published datasets for discovering functional information to support the prediction and understanding of clinical BRD.


Subject(s)
Bovine Respiratory Disease Complex/genetics , Computational Biology , Gene Expression Profiling , Gene Regulatory Networks , RNA-Seq , Supervised Machine Learning , Transcriptome , Animals , Bovine Respiratory Disease Complex/immunology , Bovine Respiratory Disease Complex/microbiology , Bovine Respiratory Disease Complex/virology , Cattle , Databases, Genetic , Host-Pathogen Interactions , Lung/immunology , Lung/microbiology , Lung/virology
16.
Am J Vet Res ; 82(9): 746-751, 2021 Sep.
Article in English | MEDLINE | ID: mdl-34432517

ABSTRACT

OBJECTIVE: To determine anti-bovine respiratory syncytial virus (BRSV) antibody titers for nasal secretions and serum from beef calves following administration of a modified-live (MLV) BRSV vaccine. ANIMALS: 60 healthy newborn purebred beef calves. PROCEDURES: Calves were randomly assigned to 1 of 3 groups: intranasal (IN)-SC (IN MLV BRSV vaccine within 24 hours of birth and SC MLV BRSV vaccine at 2 months of age), SC-IN (SC MLV BRSV vaccine within 24 hours of birth and IN MLV BRSV vaccine at 2 months of age), or NO-IN (no vaccine within 24 hours of birth and IN MLV BRSV vaccine at 2 months of age). Nasal secretion and serum samples were collected for determination of anti-BRSV antibodies within 24 hours of birth and 2 and 6 months of age. RESULTS: Titers of anti-BRSV IgA antibodies in nasal secretions and BRSV neutralizing antibodies in serum were similar among groups at each sampling time. Within 24 hours of birth, nasal anti-BRSV IgA titers were negligible. At 2 months, mean nasal anti-BRSV IgA titers for calves in IN-SC, SC-IN, and NO-IN groups were 192.84, 224.49, and 114.71, respectively. CONCLUSIONS AND CLINICAL RELEVANCE: Concentrations of anti-BRSV IgA antibodies in the nasal secretions and BRSV neutralizing antibodies in the serum of young beef calves following an MLV BRSV vaccine protocol that consisted of IN or SC vaccine within 24 hours of birth and vice versa at 2 months of age were not different from that following only an IN MLV BRSV vaccine at 2 months of age. However, the lack of any differences may have been attributed to other factors.


Subject(s)
Cattle Diseases , Respiratory Syncytial Virus Infections , Respiratory Syncytial Virus Vaccines , Respiratory Syncytial Virus, Bovine , Animals , Antibodies, Neutralizing , Cattle , Immunoglobulin A , Respiratory Syncytial Virus Infections/prevention & control , Respiratory Syncytial Virus Infections/veterinary
17.
Vet Immunol Immunopathol ; 237: 110254, 2021 Jul.
Article in English | MEDLINE | ID: mdl-34034143

ABSTRACT

This study was performed to elucidate whether the route of booster vaccination affects the immune response against respiratory vaccine viruses in pre-weaning beef calves that receive primary intranasal (IN) vaccination during the first month of life. The objective was to compare the serum neutralizing antibody (SNA) titers to BHV1, BRSV, and BPI3V, cytokine mRNA expression and mucosal BHV1- and BRSV-specific IgA in nasal secretions following administration of IN or subcutaneous (SC) modified-live virus (MLV) booster vaccines 60 days after primary IN vaccination in young beef calves. Twenty-one beef calves were administered 2 mL of an IN MLV vaccine containing BHV1, BRSV, and BPI3V (Inforce3®) between one and five weeks of age. Sixty days after primary vaccination, calves were randomly assigned to one of two groups: IN-MLV (n = 11): Calves received 2 mL of the same IN MLV vaccine used for primary vaccination and 2 mL of a SC MLV vaccine containing BVDV1 & 2 (Bovi- Shield GOLD® BVD). SC-MLV (n = 10): Calves were administered 2 mL of a MLV vaccine containing, BHV1, BRSV, BPI3V, and BVDV1 & 2 (Bovi-Shield GOLD® 5). Blood and nasal secretion samples were collected on days -61 (primary vaccination), -28, -14, 0 (booster vaccination), 14, 21, 28, 42 and 60 for determination of SNA titers, cytokine gene expression analysis and nasal virus-specific IgA concentrations. Statistical analysis was performed using a repeated measures analysis through PROC GLIMMIX of SAS®. Booster vaccination by neither IN nor SC routes induced a significant increase in SNA titers against BHV1, BRSV, and BPI3V. Subcutaneous booster vaccination induced significantly greater BRSV-specific SNA titers (on day 42) and IgA concentration in nasal secretions (on days 21 and 42) compared to calves receiving IN booster vaccination. Both IN and SC booster vaccination were able to stimulate the production of BHV1-specific IgA in nasal secretions. In summary, booster vaccination of young beef calves using either SC or IN route two months after IN MLV primary vaccination resulted in comparable SNA titers, cytokine gene expression profile and virus-specific IgA concentration in nasal secretions. Only a few differences in the systemic and mucosal immune response against BHV1 and BRSV were observed. Subcutaneous booster vaccination induced significantly greater BRSV-specific SNA and secretory IgA titers compared to IN booster vaccination.


Subject(s)
Cattle Diseases/immunology , Respiratory Syncytial Virus Vaccines/immunology , Respiratory Syncytial Virus, Bovine/immunology , Administration, Intranasal/veterinary , Animals , Animals, Newborn , Antibodies, Neutralizing/blood , Antibodies, Neutralizing/immunology , Antibodies, Viral/blood , Antibodies, Viral/immunology , Cattle , Cattle Diseases/prevention & control , Cytokines/blood , Immunization, Secondary/veterinary , Immunogenicity, Vaccine , Respiratory Syncytial Virus Vaccines/administration & dosage
18.
PLoS One ; 16(4): e0250758, 2021.
Article in English | MEDLINE | ID: mdl-33901263

ABSTRACT

BACKGROUND: Despite decades of extensive research, bovine respiratory disease (BRD) remains the most devastating disease in beef cattle production. Establishing a clinical diagnosis often relies upon visual detection of non-specific signs, leading to low diagnostic accuracy. Thus, post-weaned beef cattle are often metaphylactically administered antimicrobials at facility arrival, which poses concerns regarding antimicrobial stewardship and resistance. Additionally, there is a lack of high-quality research that addresses the gene-by-environment interactions that underlie why some cattle that develop BRD die while others survive. Therefore, it is necessary to decipher the underlying host genomic factors associated with BRD mortality versus survival to help determine BRD risk and severity. Using transcriptomic analysis of at-arrival whole blood samples from cattle that died of BRD, as compared to those that developed signs of BRD but lived (n = 3 DEAD, n = 3 ALIVE), we identified differentially expressed genes (DEGs) and associated pathways in cattle that died of BRD. Additionally, we evaluated unmapped reads, which are often overlooked within transcriptomic experiments. RESULTS: 69 DEGs (FDR<0.10) were identified between ALIVE and DEAD cohorts. Several DEGs possess immunological and proinflammatory function and associations with TLR4 and IL6. Biological processes, pathways, and disease phenotype associations related to type-I interferon production and antiviral defense were enriched in DEAD cattle at arrival. Unmapped reads aligned primarily to various ungulate assemblies, but failed to align to viral assemblies. CONCLUSION: This study further revealed increased proinflammatory immunological mechanisms in cattle that develop BRD. DEGs upregulated in DEAD cattle were predominantly involved in innate immune pathways typically associated with antiviral defense, although no viral genes were identified within unmapped reads. Our findings provide genomic targets for further analysis in cattle at highest risk of BRD, suggesting that mechanisms related to type I interferons and antiviral defense may be indicative of viral respiratory disease at arrival and contribute to eventual BRD mortality.


Subject(s)
Antiviral Agents/metabolism , Bovine Respiratory Disease Complex/pathology , Interferon Type I/metabolism , Transcriptome , Animals , Antiviral Agents/therapeutic use , Bovine Respiratory Disease Complex/drug therapy , Bovine Respiratory Disease Complex/metabolism , Bovine Respiratory Disease Complex/mortality , Cattle , Contig Mapping , Gene Expression Profiling , Male , Phenotype , Protein Interaction Maps/genetics , Toll-Like Receptor 4/metabolism
19.
Anim Health Res Rev ; 21(2): 131, 2020 12.
Article in English | MEDLINE | ID: mdl-33682662
20.
Curr Issues Mol Biol ; 42: 605-634, 2021.
Article in English | MEDLINE | ID: mdl-33627518

ABSTRACT

Vaccines can be powerful tools, but for some diseases, safe and effective vaccines have been elusive. New developments in nucleic acid sequencing, bioinformatics, and protein modeling are facilitating the discovery of previously unknown antigens through reverse vaccinology approaches. Sequencing the complementarity- determining region of antibodies and T cell receptors allows detailed assessment of the immune repertoire and identification of paratopes shared by many individuals, supporting the selection of antigens that may be broadly protective. Systems vaccinology approaches to asses the global host response to vaccination by evaluation of differentially expressed genes in blood, cellular or tissue transcriptomes can reveal previously unknown pathways and interactions related to protective immunity. While it is important to remember that discoveries made through reverse vaccinology and systems vaccinology must still be confirmed with traditional challenge models and clinical trials, these approaches can provide new perspectives that may help solve longstanding problems in veterinary vaccinology.


Subject(s)
Computational Biology , High-Throughput Nucleotide Sequencing , Models, Molecular , Proteins/chemistry , Vaccinology/methods , Animals , Complementarity Determining Regions/genetics , Complementarity Determining Regions/immunology , Computational Biology/methods , Epitopes/genetics , Epitopes/immunology , High-Throughput Nucleotide Sequencing/methods , Host-Pathogen Interactions/immunology , Humans , Proteins/immunology , Receptors, Antigen, T-Cell/genetics , Receptors, Antigen, T-Cell/immunology , Vaccines/genetics , Vaccines/immunology
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