Decision tree analysis for pathogen identification based on circumstantial factors in outbreaks of bovine respiratory disease in calves.

Respiratory tract infections continue to be a leading cause of economic loss, hampered animal welfare and intensive antimicrobial use in cattle operations, worldwide. To better target antimicrobial therapy, control and prevention towards the involved pathogens, there is a growing interest in microbiological tests on respiratory samples. However, these tests are time consuming, cost money and sampling might compromise animal welfare. Therefore, the objective of the present study was to develop immediately applicable decision trees for pathogen identification in outbreaks of bovine respiratory disease based on circumstantial factors. Data from a cross sectional study, involving 201 outbreaks of bovine respiratory disease in dairy and beef farms between 2016 and 2019 was used. Pathogens were identified by a semi-quantitative PCR (polymerase chain reaction) on a pooled non-endoscopic broncho-alveolar lavage sample from clinically affected animals. Potential risk factors of involved animals, environment, management and housing were obtained by enquiry. Classification and regression tree analysis was used for decision tree development with cross-validation. Different trees were constructed, involving a general 3-group classification tree (viruses, Mycoplasma bovis or Pasteurellaceae family) and a tree for each single pathogen. The general 3- group classification tree was 52.7 % accurate and had a sensitivity of 81.5 % and a specificity 52.2 % for viruses, respectively 51.7 % and 84.4 % for M. bovis and 28.9 % and 93.6 % for Pasteurellaceae. The single-pathogen trees were more specific than sensitive: Histophilus somni (Se = 25.8 %; Sp = 94.5 %), Mannheimia haemolytica (Se = 69.2 %; Sp = 70.6 %), bovine coronavirus (Se = 42.2 %; Sp = 89.6 %) and bovine respiratory syncytial virus (Se = 34.0 %; Sp = 96.6 %). For Pasteurella multocida, M. bovis and parainfluenzavirus type 3 no meaningful tree was obtained. The concept and trees are promising, but currently lack sensitivity and specificity in order to be a reliable tool for practice. For now, the obtained trees can already be informative for decision making to some extend depending on the end node in which an outbreak falls.

Assessment of Metagenomic Sequencing and qPCR for Detection of Influenza D Virus in Bovine Respiratory Tract Samples.

High throughput sequencing is currently revolutionizing the genomics field and providing new approaches to the detection and characterization of microorganisms. The objective of this study was to assess the detection of influenza D virus (IDV) in bovine respiratory tract samples using two sequencing platforms (MiSeq and Nanopore (GridION)), and species-specific qPCR. An IDV-specific qPCR was performed on 232 samples (116 nasal swabs and 116 tracheal washes) that had been previously subject to virome sequencing using MiSeq. Nanopore sequencing was performed on 19 samples positive for IDV by either MiSeq or qPCR. Nanopore sequence data was analyzed by two bioinformatics methods: What's In My Pot (WIMP, on the EPI2ME platform), and an in-house developed analysis pipeline. The agreement of IDV detection between qPCR and MiSeq was 82.3%, between qPCR and Nanopore was 57.9% (in-house) and 84.2% (WIMP), and between MiSeq and Nanopore was 89.5% (in-house) and 73.7% (WIMP). IDV was detected by MiSeq in 14 of 17 IDV qPCR-positive samples with Cq (cycle quantification) values below 31, despite multiplexing 50 samples for sequencing. When qPCR was regarded as the gold standard, the sensitivity and specificity of MiSeq sequence detection were 28.3% and 98.9%, respectively. We conclude that both MiSeq and Nanopore sequencing are capable of detecting IDV in clinical specimens with a range of Cq values. Sensitivity may be further improved by optimizing sequence data analysis, improving virus enrichment, or reducing the degree of multiplexing.

Risk Assessment for Use of a Porcine Circovirus-Contaminated Reagent in a Barrier Maintained Rodent Colony.

A proposal for the use of porcine pancreatic elastase (PPE) to develop a mouse model of pulmonary emphysema raised concerns about introducing contaminating porcine viruses into our barrier facility. Porcine Circovirus (PCV) is a known contaminant of vaccines and cell cultures that have been exposed to porcine-derived reagents. Endemic infection of PCV3 in laboratory mice has been reported, and some evidence supports natural PCV infection in wild mice. PPE samples from 2 different vendors tested positive for DNA from both PCV2 and 3. To allow model development with these reagents to proceed, we developed a protocol that would meet scientific objectives, minimize exposure of mice, and provide information on the potential for the virus to spread. Five d after BALB/c mice received intralaryngeal administration of PPE, lungs were harvested and analyzed for evidence of disease. Tissues from other major organs were submitted to test for disseminated PCV2 and 3 DNA. Similarly, tissues (including lungs) from direct contact nude sentinel mice were analyzed for the presence of the virus. To evaluate the possibility of endemic PCV2/3 infection, we also surveyed non-porcine reagent exposed mice on other studies. PCV2 and 3 was not detected in any of the tissues submitted. Although this study provided no evidence of infection and transmission of PCV2/3 from the contaminated PPE sample over the 5 d study, further work is needed to understand the risks and impact of introducing PCV contaminated cells or reagents into barrier maintained rodent colonies.

An estimation of US horse-owner/caregiver willingness-to-pay for daily use and infectious upper respiratory disease treatment options.

BACKGROUND: Equine injury and disease cause two types of costs for those financially responsible for treating and caring for the infected horse(s); direct costs of treating the horse and indirect cost of lost use of the horse for a period of time to the user of the horse (daily horse use). Indirect costs are more difficult to estimate but pose significant financial implications for equine-owners/caregivers. Additionally, there exists a gap in existing research regarding the valuation of infectious treatment options in horses. OBJECTIVE: To estimate the value a US horse-owner/caregiver places on daily horse use and describe respondents' willingness-to-pay for various attributes of equine treatment options. STUDY DESIGN: Online questionnaire survey. METHODS: An online questionnaire was provided to equine-owners and caretakers, and owner demographic, horse care and horse use information from respondents were requested. Additionally, respondents were presented with hypothetical disease treatment options with the following attributes: daily dosage, number of days of rest required, route of administration and out-of-pocket cost to the owner/caretaker through a choice experiment. Data were analysed using a rank-ordered logit analysis and willingness-to-pay estimates for daily use and treatment options were calculated. RESULTS: Results suggest that the average horse-owner with an uninsured and insured horse is willing to pay $12.07 (95% confidence interval: -$15.01, -$9.69) and $17.95 (95% confidence interval: -$25.30, -$11.20) per day to reduce lost use days required (due to need for rest) respectively. Respondents showed preferences for oral administration over treatments requiring i.m. injections. MAIN LIMITATIONS: As this study employed an online survey it was subjected to self-selection bias and a sample size calculation was not performed. CONCLUSIONS: Veterinarians and pharmaceutical companies may use these results when promoting various treatment options to horse-owners/caregivers and in product development. Additionally, promotion efforts may be targeted towards equine-owners with higher daily use values (owners with insured horses).

Dexamethasone treatment differentially alters viral shedding and the antibody and acute phase protein response after multivalent respiratory vaccination in beef steers.

Our objective was to examine immunosuppression induced by dexamethasone (DEX) administration in cattle on immunological responses to a multivalent respiratory vaccine containing replicating and nonreplicating agents. Steers ( = 32; 209 ± 8 kg) seronegative to infectious bovine rhinotracheitis virus (IBRV), bovine viral diarrhea virus (BVDV), bovine respiratory syncytial virus (BRSV), and parainfluenza-3 virus (PI3V) were stratified by BW and randomly assigned to 1 of 3 treatments: 1) acute immunosuppression (ACU; 0.5 mg/kg BW DEX intravenously at 1000 h only on d 0), 2) chronic immunosuppression (CHR; 0.5 mg/kg BW DEX intravenously at 1000 h on d -3 to 0), or 3) a control (CON; no DEX). On d -4, steers were fitted with intravenous catheters in the jugular vein and placed into individual stanchions. At 1200 h on d 0, steers were administered a respiratory vaccine containing modified-live virus (MLV) isolates of IBRV, BVDV, BRSV, and PI3V and a (MH) toxoid. On d 4, cattle were transported (177 km) and housed in an isolated outdoor pen. Serum was harvested on d 0, 7, 14, 21, 28, 35, 42, and 56 to determine IBRV-, BVDV-, BRSV-, and PI3V-specific antibody titers and MH whole cell and leukotoxin antibody concentrations. Sera from d -2, 0, 1, 3, 7, and 14 were used to quantify haptoglobin (Hp) concentration and ceruloplasmin (Cp) activity. Nasal swab specimens were collected on d 0, 3, and 14 to determine the presence of IBRV, BVDV, BRSV, and PI3V via PCR analysis. There was a treatment × day interaction ( < 0.01) such that CHR steers had a greater ( ≤ 0.07) BVDV antibody titer on d 14, 21, and 28. Moreover, IBRV-specific antibodies increased beginning on d 14 for CHR and on d 28 for ACU and remained greater through d 56 compared with CON ( ≤ 0.03). Conversely, serum MH whole cell antibody concentration was least ( ≤ 0.06) for CHR from d 7 to 28 and greatest for CON ( ≤ 0.04) on d 56. Treatment altered Hp such that CON exhibited a greater ( < 0.01) Hp concentration than CHR but was not different from ACU ( = 0.16). On d 3, Cp was greatest for CON, intermediate for ACU, and least for CHR (treatment × day; ≤ 0.01). The prevalence of IBRV and BVDV in nasal swabs on d 14 was 67 and 56%, respectively, for CHR; 10 and 10%, respectively, for CON; and 9 and 0%, respectively, for ACU ( ≤ 0.006). Results suggest that CHR allowed increased replication of MLV vaccine agents. Conversely, DEX-induced immunosuppression blunted the acute phase protein and antibody response against the nonreplicating MH toxoid.

Association between a shelter-neuter-return program and cat health at a large municipal animal shelter.

OBJECTIVE To determine effects of a shelter-neuter-return (SNR) program on cat admissions and health at a large municipal animal shelter in Northern California. DESIGN Retrospective cohort study. ANIMALS 117,383 cats for which data were recorded in the San Jose Animal Care Center database between January 1, 2006, and December 31, 2013. PROCEDURES Shelter records were analyzed for trends in cat demographic data, shelter intake and outcome types, and prevalence of upper respiratory infection (URI) over the 8-year period and before and after initiation of an SNR program on March 8, 2010. RESULTS Number of cats admitted to the shelter each year decreased significantly over 8 years; beginning in 2010, duration of stay decreased. Proportion of cats euthanized decreased from 66.6% (28,976/43,517) in the pre-SNR period to 34.9% (11,999/34,380) in the post-SNR period, whereas prevalence of URI increased from 5.5% to 6.8%, and median duration of shelter stay decreased from 6 to 5 days for cats < 4 months of age and from 8 to 6 days for older cats. With implementation of the SNR program and a new treatment policy for cats with URI, more cats received treatment with less medication, yielding cost savings. CONCLUSIONS AND CLINICAL RELEVANCE Initiation of the SNR program was associated with a decreased number of cats admitted to the shelter and a lower proportion euthanized. With increased resources to care for cats with URI and changes in the URI treatment protocol, fewer cats were euthanized for URI and more cats were treated at lower cost and with a briefer shelter stay.

Development of an EvaGreen-based multiplex real-time PCR assay with melting curve analysis for simultaneous detection and differentiation of six viral pathogens of porcine reproductive and respiratory disorder.

Concurrent infection of pigs with two or more pathogens is common in pigs under intensive rearing conditions. Porcine circovirus type 2 (PCV2), porcine parvovirus (PPV), porcine reproductive and respiratory syndrome virus (PRRSV), classical swine fever virus (CSFV), Japanese encephalitis virus (JEV) and pseudorabies virus (PRV) are all associated with reproductive or respiratory disorders or both and can cause significant economic losses in pig production worldwide. An EvaGreen-based multiplex real-time PCR (EG-mPCR) with melting curve analysis was developed in this study for simultaneous detection and differentiation of these six viruses in pigs. This method is able to detect and distinguish PCV2, PPV, PRRSV, CSFV, JEV and PRV with the limits of detection ranging from 100 to 500 copies/µL, high reproducibility, and intra-assay and inter-assay variation ranging from 0.11 to 3.20%. After validation, a total of 118 field samples were tested by the newly developed EG-mPCR. PCV2 was identified in 23%, PPV in 15%, PRRSV in 17% and PRV in 5% of the samples. Concurrent PCV2 and PRRSV infection was detected in 6.7%, PCV2 and PPV in 5% and PPV2 and PRRSV infection was detected in 5% of the cases. The agreement of the EG-mPCR and conventional PCR tests was 99.2%. This EG-mPCR will be a useful, rapid, reliable and cost-effective alternative for routine surveillance testing of viral infections in pigs.

Prevalence of upper respiratory pathogens in four management models for unowned cats in the Southeast United States.

Upper respiratory infection (URI) is a pervasive problem in cats and impacts the capacity and cost of sheltering programs. This study determined the pattern of respiratory pathogens in cats with and without clinical signs of URI in four different models for managing unowned cats, namely, (1) short-term animal shelters (STS), (2) long-term sanctuaries (LTS), (3) home-based foster care programs (FCP), and (4) trap-neuter-return programs for community cats (TNR). Conjunctival and oropharyngeal swabs from 543 cats, approximately half of which showed clinical signs of URI, were tested for feline herpes virus-1 (FHV), feline calicivirus (FCV), Chlamydia felis, Bordetella bronchiseptica, Mycoplasma felis, and canine influenza virus by real-time PCR. FHV (59%, 41%) and B. bronchiseptica (33%, 24%) were more prevalent in both clinically affected and nonclinical cats, respectively, in STS than other management models. FCV (67%, 51%) and M. felis (84%, 86%) were more prevalent in LTS than any other management model. Clinically affected cats in FCP were more likely to carry FHV (23%, 6%), C. felis (24%, 10%), or M. felis (58%, 38%) than were nonclinical cats. Clinically affected cats in TNR were more likely to carry FCV (55%, 36%) or C. felis (23%, 4%) than were nonclinical cats. The prevalence of individual pathogens varied between different management models, but the majority of the cats in each model carried one or more respiratory pathogens regardless of clinical signs. Both confined and free-roaming cats are at risk of developing infectious respiratory disease and their health should be protected by strategic vaccination, appropriate antibiotic therapy, effective biosecurity, feline stress mitigation, and alternatives to high-density confinement.

Intervention strategies for emerging respiratory virus infections: policy and public health considerations.

Respiratory viruses have emerged and re-emerged in humans for hundreds of years. In the recent past avian and animal influenza viruses have caused human disease ranging from conjunctivitis to respiratory illnesses, including the 2009-10 A(H1N1)pdm09 pandemic. Coronaviruses, human metapneumovirus (hMPV) and enteroviruses have also impacted humans globally. Since the likely public health impacts are common, plans and policies for intervention strategies can be developed, encompassing early detection through surveillance and diagnostics, as well as treatment and prevention through clinical and non-clinical interventions. The global comprehensiveness of these varies according to differing resources, competing health priorities and the causative agent, yet, irrespective of this, activities must be proportional to the threat. Pandemics and severe epidemics enable policies to be tested and gaps identified.

Microbiologic and cytologic assessment of bronchoalveolar lavage fluid from dogs with lower respiratory tract infection: 105 cases (2001-2011).

BACKGROUND: Documentation of lower respiratory tract infection has relied on microbiologic and cytologic findings in airway fluid, but there is no gold standard for making a definitive diagnosis. OBJECTIVE: To report cytologic and microbiologic findings in dogs diagnosed with lower respiratory tract infection through evaluation by bronchoscopy and bronchoalveolar lavage. ANIMALS: A total of 105 dogs with spontaneous respiratory disease. METHODS: Retrospective case review of all dogs identified through the electronic medical record database that had bronchoscopy with bronchoalveolar lavage performed between 2001 and 2011. Results of bronchoalveolar lavage cytology and microbiology were evaluated in 510 dogs, and 105 cases with septic, suppurative inflammation or bacterial growth from cultures were examined further. RESULTS: Bacteria were isolated from 89/105 aerobic cultures, 18/104 anaerobic cultures, and 30/99 Mycoplasma spp. cultures. The most common isolate was Mycoplasma spp. followed by Pasteurella sp., Bordetella sp, Enterobacteriaceae, and anaerobes. A single bacterial species was cultured from 44/99 dogs (44%) and multiple bacterial species were isolated from 55/99 dogs (56%). Suppurative inflammation with intracellular bacteria was identified cytologically in 78 of 105 dogs (74%). In 27 dogs that lacked cytologic evidence of sepsis, mixed (n = 18) and neutrophilic (n = 9) inflammation was reported, and Mycoplasma spp. (13/27) or Bordetella spp. (7/27) were most commonly isolated. Most aerobic bacteria were susceptible to routinely used antimicrobial drugs. CONCLUSIONS AND CLINICAL IMPORTANCE: Confirmation of lower respiratory tract infection in dogs is challenging and organisms can be isolated from dogs in which bacteria are not detected on cytologic examination.

Assessment of oxidative stress biomarkers in exhaled breath condensate and blood of Thoroughbred foals.

This study reports preliminary data on systemic and local biomarkers of oxidative stress (OS) in Thoroughbred foals. Blood and exhaled breath condensate (EBC) were sampled from 13 foals on two farms. Values of reactive oxygen metabolites (d-ROMs), biological antioxidant potential (BAP), antioxidant barrier (OXY-ads), thiol antioxidant barrier (SHp) and advanced oxidation protein products (AOPPs) were determined in blood, while EBC samples were assayed for hydrogen peroxide (H(2)O(2)) levels. Systemic and local OS biomarkers did not differ between farms and gender. Increased oxidative stress index (OSI) values in a foal recovering from pneumonia and elevated H(2)O(2) in EBC coupled with low SHp and elevated AOPPs in the blood of a foal with overt upper respiratory tract disease suggested that the OS markers measured in this study may relate to the respiratory health status of foals.

A retrospective analysis of factors correlated to chimpanzee (Pan troglodytes schweinfurthii) respiratory health at Gombe National Park, Tanzania.

Infectious disease and other health hazards have been hypothesized to pose serious threats to the persistence of wild ape populations. Respiratory disease outbreaks have been shown to be of particular concern for several wild chimpanzee study sites, leading managers, and researchers to hypothesize that diseases originating from and/or spread by humans pose a substantial risk to the long-term survival of chimpanzee populations. The total chimpanzee population in Gombe National Park, Tanzania, has declined from 120-150 in the 1960s to about 100 by the end of 2007, with death associated with observable signs of disease as the leading cause of mortality. We used a historical data set collected from 1979 to 1987 to investigate the baseline rates of respiratory illness in chimpanzees at Gombe National Park, Tanzania, and to analyze the impact of human-related factors (e.g., banana feeding, visits to staff quarters) and non-human-related factors (e.g., sociality, season) on chimpanzee respiratory illness rates. We found that season and banana feeding were the most significant predictors of respiratory health clinical signs during this time period. We discuss these results in the context of management options for the reduction of disease risk and the importance of long-term observational data for conservation.

Effects of exposure to calves persistently infected with bovine viral diarrhea virus type 1b and Mannheimia haemolytica challenge on animal performance, nitrogen balance, and visceral organ mass in beef steers.

Bovine viral diarrhea viruses (BVDV) have been isolated alone or in combination with other viral and bacterial pathogens in animals diagnosed with bovine respiratory disease (BRD), a disease causing major economic loss to the feedlot industry. The objective of this experiment was to determine the effects of Mannheimia haemolytica challenge after short-term exposure (72 h) to bovine viral diarrhea virus type 1b (BVDV1b) persistently infected (PI) calves on performance, N balance, and organ mass in finishing cattle. Treatments (6 steers/treatment; initial BW = 314 +/- 31 kg) were 1) steers not exposed to steers PI with BVDV nor challenged with M. haemolytica (control; CON); 2) steers exposed to 2 steers PI with BVDV1b (BVD) for 72 h; 3) steers intratracheally challenged with M. haemolytica (MH); or 4) steers exposed to 2 steers PI with BVDV1b for 72 h and challenged with M. haemolytica (BVD+MH). There were 12 h between exposure to PI steers and challenge with M. haemolytica. Steers were housed in metabolism stanchions during the first 5 d after the M. haemolytica challenge and on d 7 to 11, 28 to 32, and for 5 d before slaughter (average 119 d on feed) to determine N balance and were weighed every 28 d. At slaughter, carcass and organ mass data were collected. Data were analyzed as a randomized complete block design with a 2 x 2 factorial arrangement of treatments, and steer was used as the experimental unit. From d -3 (beginning of PI steer exposure) to 4, steers challenged with M. haemolytica had less (P = 0.04) ADG than steers not challenged with M. haemolytica. In addition, steers exposed to steers PI with BVDV tended (P = 0.09) to have less ADG and G:F across the entire finishing period than steers not exposed to BVDV. Before slaughter, retained N expressed as grams per day (P = 0.03) and as a percentage of N intake (P = 0.04) was less in BVD steers compared with steers not exposed to BVDV. There were no effects (P > 0.10) of BVDV exposure or M. haemolytica challenge on empty BW (EBW) or carcass characteristics. Expressed as a percentage of EBW, HCW was less (P = 0.02) and total offal weight was greater (P = 0.02) for steers challenged with M. haemolytica compared with steers not challenged. Results are in agreement with those reported in larger scale finishing studies and suggest that acute exposure to BRD-related pathogens can have long-term effects on animal performance.

Effects of exposure to calves persistently infected with bovine viral diarrhea virus type 1b and subsequent infection with Mannheima haemolytica on clinical signs and immune variables: model for bovine respiratory disease via viral and bacterial interaction.

The objective was to determine effects of an intratracheal Mannheimia haemolytica challenge after 72-h exposure to bovine viral diarrhea virus type 1b (BVDV1b) persistently infected (PI) calves on serum antibody production, white blood cell count (WBC), cytokine concentrations, and blood gases in feedlot steers. Twenty-four steers (initial BW = 314 +/- 31 kg) were randomly allocated to 1 of 4 treatments (6 steers/treatment) arranged as a 2 x 2 factorial. Treatments were 1) steers not exposed to steers PI with BVDV nor challenged with M. haemolytica (control; CON); 2) steers exposed to 2 steers PI with BVDV for 72 h (BVD); 3) steers intratracheally challenged with M. haemolytica (MH); and 4) steers exposed to 2 steers PI with BVDV for 72 h and challenged with M. haemolytica (BVD+MH). There were 12 h between exposure to PI steers and challenge with M. haemolytica. Rectal temperature was increased (P < 0.001) for MH and BVD+MH during the initial 24 h after the M. haemolytica challenge. For MH and BVD+MH, total WBC count was increased (P < 0.01) at 36 h post M. haemolytica challenge compared with CON, whereas in BVD steers, WBC count was decreased (P < 0.01). Total lymphocyte count was increased (P = 0.004) during the initial 72 h post BVDV exposure for the BVD and BVD+MH groups compared with MH and CON, and this difference remained at 96 h post M. haemolytica challenge. An increased (P < 0.001) total neutrophil count was observed during the initial 36 h for the MH group and at 72 h for the BVD+MH challenge group. Interleukin 1beta, IL-6, and tumor necrosis factor alpha (TNFalpha) concentrations were greater (P

Calf health monitoring in Norwegian dairy herds.

The aims of this study were to estimate the incidence of calf diseases in Norwegian dairy herds, improve calf health recordings in the Norwegian Cattle Health Recording System (NCHRS), and investigate different methods for validation of calf health data. A longitudinal, cross-sectional survey of calf health in Norway was performed between September 1, 2004, and January 31, 2007. The participating dairy herds were randomly selected from among herds registered in the Norwegian Dairy Herd Recording System as having at least 15 cow-years. Each herd participated for 1 yr. Diseases and treatments of calves of up to 180 d of age in 135 dairy herds were reported using the NCHRS. In total, 6,668 calves were born in the participating dairy herds during the project period. A total of 573 (29.6%) of the 1,936 calf health recordings reported were recordings of diseases and 1,363 (70.4%) were events of preventive therapy, dehorning, or castration. The recorded incidence of diarrhea and respiratory disease was 3.8% and 2.9%, respectively. The median age of occurrence of diarrhea and respiratory disease was 17 and 37 d, respectively. Three different methods, based on sampling of diseased calves, dehorning as an indicator of a well-functioning recording system, or feedback on degree of commitment to calf health recording, were tested to assess validation of the calf health records. The 3 methods indicated an underestimation of calf health records in the NCHRS of approximately 40% and an estimated "true" incidence of diarrhea and respiratory disease of 5.5 and 4.1%, respectively. The results from this study demonstrate the importance of encouraging farmers to conduct calf health recordings. They also indicate that finding a standardized method for validation of health data is a considerable challenge.

Comparative assessment of a metabolically attenuated Mycoplasma gallisepticum mutant as a live vaccine for the prevention of avian respiratory mycoplasmosis.

In a previous study, signature sequence mutagenesis (SSM) was used to identify a mutant with a disruption of the gene encoding the metabolic factor, dihydrolipoamide dehydrogenase, and that mutant was designated Mg 7. The current study assessed the safety, immunogenicity and efficacy of Mg 7 in comparison to two commercially available vaccines (ts-11 and F) as well as a laboratory vaccine strain, GT5. Intratracheal vaccination of chickens with all four attenuated mutants induced varying levels of protection against intratracheal challenge with virulent Mycoplasma gallisepticum strain R(low). Mg 7 vaccinated chickens rapidly cleared the challenge strain, had lower histopathologic tracheal lesion scores when compared to unvaccinated chickens, and mounted a strong humoral anti-M. gallisepticum-specific IgG response. The IgG levels increased 2- to 3-fold upon R(low) challenge. Mg 7 induced a greater level of protection against intratracheal R(low) challenge than that observed with the other three attenuated strains, as evidenced by a lower recovery of R(low) from tracheas and lower histopathologic lesion scores in tracheas and air sacs. Based on these findings, Mg 7 appears to have good potential as a safe and effective vaccine for the prevention of avian mycoplasmosis.

[The importance of Mycoplasma bovis in bovine respiratory disease]. / Het belang van Mycoplasma bovis bij luchtwegaandoeningen van kalveren.

The annual damage caused by bovine respiratory disease is estimated at 45 up to 55 euro per calf of milking cattle and 117.50 euro per veal calf In Europe, M. bovis is responsible for at least 1/4 to 1/3 of all pneumonia cases in calves. Serology may help to identify the spreading of these bacteria in a herd.

Assessment of neutrophil components as markers of lung injury in the course of bovine respiratory tract infections.

To define the role of activated neutrophils in lung injury during bovine respiratory tract infections (BRTI) their in vitro function was investigated. As a means to achieve this goal the comparison of secretory action between neutrophils from the BRTI group and control was made on the basis of elastase, myeloperoxidase (MPO), alkaline phosphatase (ALK-P) release, and nitric oxide production. We noted that there is an interdependence between secretory response of neutrophils and clinical severity of BRTI. The release of elastase was greater in the BRTI group than in the control group (49.17+/-4.41 versus 46.43+/-4.95% of the total content). Neutrophils from infected heifers exhibited a significantly (p<0.05) higher value of MPO release than from healthy heifers and reached 39.23+/-10.18 versus 25.54+/-8.41% of the total content. ALK-P containing granules released significantly (p<0.001) more enzyme in the group with BRTI than in the control group (22.42+/-6.27 versus 13.74+/-2.01% of the total enzyme content). The level of nitrite accumulation rose in the culture of cells isolated from heifers with BRTI from 4+/-0.53 microM after 0.5h to 6.9+/-0.52 microM after 72 h. Our data suggest that during BRTI the increase of neutrophil secretory action results in augmentation of enzyme release including elastase, MPO and ALK-P, and the nitrite production. During an excessive secretory response of neutrophils all these factors contribute to lung injury and worsen the course of a disease and might be recognised as markers of lung injury. Moreover, such a destructive action of neutrophils must be taken into account during the introduction of new methods of BRTI treatment.