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JDS Communications May 2023Appropriate sample collection, storage conditions, and time for transport to the laboratory are important for an accurate diagnostic result. We evaluated the effects of...
Appropriate sample collection, storage conditions, and time for transport to the laboratory are important for an accurate diagnostic result. We evaluated the effects of transport storage medium type, time of storage, and storage temperatures on (MH) and (PM) recovery using an in vitro model simulation. A quantitative culture method, using colony-forming units per milliliter, was used to recover MH or PM by an in vitro model with cotton swabs. Three independent trials were conducted, in which cotton swabs were inoculated with MH or PM and placed in either (1) a sterile 15-mL polypropylene tube without transport medium (dry), (2) Amies culture medium with charcoal (ACM), or (3) Cary-Blair transport agar (CBA). Swabs were evaluated for recovery of MH or PM when stored at 3 temperatures (4°C, 23°C, or 36°C) and after storage for 8 h, 24 h, or 48 h. From all study group combinations, a total of 162 individual independent swabs were evaluated. The nonparametric Dunn all-pairs approach was used to compare the proportion of culturable bacteria, between the various storage media, temperature, and time point combinations. The proportion of MH in samples stored at 4°C was significantly higher for ACM and CBA than dry storage at 24 and 48 h. The MH samples stored at 36°C had a significantly higher proportion for ACM and CBA than dry storage at 24 h. The proportion of PM in samples stored at 4°C was significantly lower for ACM compared with dry at 8 h but significantly higher at 48 h. The PM samples stored at 23°C in ACM had a significantly higher proportion than dry samples at 24 h, and, at 48 h, ACM and CBA had a significantly higher proportion than the dry group. All swabs stored at 36°C for 48 h had a proportion close to zero, indicating decreasing diagnostic efficacy. These results support the use of transport media such as ACM and CBA for increasing the detection of PM and MH from samples, especially when samples are exposed to high temperatures. The combination of longer periods from collection of samples to diagnostic evaluation (>24 h) and higher storage temperatures (>23°C) were shown to significantly impair diagnostic accuracy.
PubMed: 37360122
DOI: 10.3168/jdsc.2022-0329 -
Scientific Reports Jun 2023Respiratory diseases caused by Mannheimia haemolytica (M. haemolytica) and Pasteurella multocida (P. multocida) have been known to result in a considerable loss due to...
Identification of serotypes of Mannheimia haemolytica and Pasteurella multocida from pneumonic cases of sheep and goats and their antimicrobial sensitivity profiles in Borana and Arsi zones, Ethiopia.
Respiratory diseases caused by Mannheimia haemolytica (M. haemolytica) and Pasteurella multocida (P. multocida) have been known to result in a considerable loss due to mortality and reduced production. This study aimed at isolation and identification of M. haemolytica and P. multocida associated with pneumonic pasteurellosis in sheep and goats using bacteriological and molecular techniques. Identification of serotypes of M. haemolytica and P. multocida was done using indirect haemagglutination test. The in vitro antimicrobial sensitivity profiles of the M. haemolytica were tested using standard disk diffusion method. A total of 52 and 78 nasal swabs were collected from pneumonic cases for bacterial isolation and identification in Borana and Arsi zone, respectively. Four hundred sera samples were collected for identification of serotypes. The results showed that 17 of 52 (32.69%; 95% CI 20.33, 47.11) nasal swabs collected from pneumonic animals in Borana yielded positive results for Pasteurella/Mannheimia species, 13 (25.00%; 95% CI 14.03, 38.95) of which were M. haemolytica. None of the samples yielded P. multocida. Twenty-three of 78 (29.49%; 95% CI 19.69, 40.89) nasal swabs collected at Arsi from pneumonic animals yielded positive results for M. haemolytica (17) and P. multocida (6). Secondary biochemical characterization revealed that 14 of the 17 isolates conform to M. haemolytica whereas none of the 6 isolates suspected to be P. mutocida were confirmed. Eleven (84.62%) isolates from Borana and 4 (28.57%) from Arsi were confirmed to be M. haemolytica using PCR targeting the Rpt2 genes. Assay for M. haemolytica serotype A1 revealed all belong to A1. None of the isolates with cultural and morphological features of P. multocida gave positive results by molecular assay. Serological assay identified three serotypes of M. haemolytica namely A1, A2 and A7 almost in all of the samples whereas P. multocida serotype A was detected in 78.75% of the samples. The M. haemolytica isolates tested for susceptibility to antibiotics showed resistance against Bacitracin (83.33%) and Penicillin (50.00%) while they were found susceptible to Gentamycin (100%), Chloramphenicol (100%) and Sulfamethoxazole (100%) and Tetracycline (83.33%). In conclusion, the results of the present study revealed the association of M. haemolytica with pneumonic pasteurellosis in sheep and goats and can be of use in vaccine development in Ethiopia. Nevertheless, further investigations and continuous monitoring of antimicrobial resistance and appropriate selection and prudent use of antimicrobials in livestock sector are required.
Topics: Cattle; Animals; Sheep; Mannheimia haemolytica; Pasteurella multocida; Pasteurellosis, Pneumonic; Serogroup; Ethiopia; Goats; Pasteurella; Anti-Bacterial Agents; Sheep Diseases
PubMed: 37268660
DOI: 10.1038/s41598-023-36026-2 -
Veterinary Medicine and Science Jul 2023Small ruminants are the most numerous of man's domestic livestock. Although sheep represent a great resource for Ethiopia, the net rate of productivity per animal is...
Isolation and identification of Mannheimia haemolytica and Pasteurella multocida from symptomatic and asymptomatic sheep and their antibiotic susceptibility patterns in three selected districts of north Gondar zone, Gondar Ethiopia.
BACKGROUND
Small ruminants are the most numerous of man's domestic livestock. Although sheep represent a great resource for Ethiopia, the net rate of productivity per animal is very low due to many factors including respiratory disorders.
OBJECTIVES
The objectives of this work were to isolate and identify M. haemolytica and P. multocida as well as to assess the antibiotic susceptibility patterns of these isolates. Nasal swab samples were collected aseptically by using 70% alcohol as a disinfectant.
METHODS
A cross-sectional study was conducted in three selected districts of the north Gondar zone, Ethiopia.
RESULTS
From 148 samples collected in 94 (63.5%) asymptomatic and 54 (35.5%) symptomatic sheep, a total of 23 were isolated successfully based on cultural, staining, and biochemical characteristics. Of these isolates, 18 (78.3%) and 5 (21.7%) were M. haeimolytica and P. multocida, respectively. Compared with the total animals examined, the proportion of M. haeimolytica and P. multocida were 12.16 % (n = 18) and 3.38% (n = 5), respectively. All of the isolates were subjected to a panel of 8 antibiotic discs for sensitivity testing. Of the tested antibiotics, chloramphenicol (100%), gentamicin, and tetracycline (82.6%) each and co-trimoxazole (60.8%) were found to be the most effective drugs whereas, both species were completely resistant to vancomycin and showed a very low degree of susceptibility for the rest drugs.
CONCLUSIONS
In conclusion, M. haemolytica was found to be the predominant isolate in all host-related factors and most of the antibiotics were not fully effective against the isolates. Hence, treatment and/or vaccination of ovine pneumonic pasteurellosis should be emphasised to M. haeimolytica using the most effective drugs along with appropriate herd management practices.
Topics: Sheep; Animals; Mannheimia haemolytica; Pasteurella multocida; Ethiopia; Cross-Sectional Studies; Anti-Bacterial Agents
PubMed: 37197762
DOI: 10.1002/vms3.1166 -
Animals : An Open Access Journal From... May 2023is one of the major causes of bovine respiratory disease in cattle. The organism is the primary bacterium isolated from calves and young cattle affected with enzootic...
is one of the major causes of bovine respiratory disease in cattle. The organism is the primary bacterium isolated from calves and young cattle affected with enzootic pneumonia. Novel indirect ELISAs were developed and evaluated to enable quantification of antibody responses to whole cell antigens using A1 strain P1148. In this study, the ELISAs were initially developed using sera from both -culture-free and clinically infected cattle, then the final prototypes were tested in the validation phase using a larger set of known-status sera ( = 145) collected from feedlot cattle. The test showed good inter-assay and intra-assay repeatability. Diagnostic sensitivity and specificity were estimated at 91% and 87% for IgG at a cutoff of S/P ≥ 0.8. IgM diagnostic sensitivity and specificity were 91% and 81% at a cutoff of sample to positive (S/P) ratio ≥ 0.8. IgA diagnostic sensitivity was 89% whereas specificity was 78% at a cutoff of S/P ≥ 0.2. ELISA results of all isotypes were related to the diagnosis of respiratory disease and isolation of (-value < 0.05). These data suggest that ELISAs can be adapted to the detection and quantification of antibody in serum specimens and support the use of these tests for the disease surveillance and disease prevention research in feedlot cattle.
PubMed: 37174567
DOI: 10.3390/ani13091531 -
Vaccine May 2023Bovine respiratory disease is the greatest threat to calf health. In this study, colostrum-fed dairy X beef calves were vaccinated at ∼30 days of age with an...
Bovine respiratory disease is the greatest threat to calf health. In this study, colostrum-fed dairy X beef calves were vaccinated at ∼30 days of age with an adjuvanted parenteral vaccine containing modified live bovine viral diarrhea virus (BVDV) type 1 and type 2, bovine herpesvirus 1 (BHV-1), bovine parainfluenza type 3 virus (PI3V) and bovine respiratory syncytial virus (BRSV) andM. haemolyticatoxoid (Group 1), or intranasal temperature-sensitive BHV-1, BRSV and PI3V concurrently witha parenteral vaccine containing modified live BVDV type 1 and type 2 andM. haemolyticatoxoid (Group 2) or a placebo (Group 3). The calves were challenged ∼150 days post vaccination intranasally with BVDV 1b and then 7 days later intratracheally withM. haemolytica. The calves wereeuthanized 6 days after theM. haemolyticachallenge. Clinical signs following BVDV infection were similar in all groups. There was increased rectal temperatures in the Groups 2 and 3 on day 3 and in Group 3 on days 8-13. Group 1 animals had a slight leukopenia following BVDV infection while Groups 2 and 3 had greater leukopenia. BVDV type 1 and 2 serum titers increased in Group 1 following vaccination while these titers waned in Groups 2 and 3. There were higher levels of BVDV in the buffy coats and nasal samples in Group 2 and Group 3 versus Group 1 (p < 0.01). Interferon-gamma response was higher (p < 0.01) in Group 1 animals than Groups 2 and 3. Group 1 had the lowest percent pneumonic tissue (1.6%) while Group 2 vaccinates had 3.7% and the control Group 3 was 5.3%. Vaccination in the face of maternal antibody with a parenteral adjuvanted vaccine resulted in better protection than the regimen of an intranasal vaccine anda parenteral adjuvanted BVDV andM haemolyticacombination vaccine in a BVDV-M. haemolyticadual challenge.
Topics: Animals; Cattle; Bovine Virus Diarrhea-Mucosal Disease; Antibodies, Viral; Diarrhea Viruses, Bovine Viral; Cattle Diseases; Diarrhea Virus 1, Bovine Viral; Vaccination; Respiratory Tract Diseases; Herpesvirus 1, Bovine; Diarrhea; Leukopenia; Mannheimia; Viral Vaccines
PubMed: 37045678
DOI: 10.1016/j.vaccine.2023.04.005 -
Veterinary World Jan 2023causes respiratory infection and mortality in sheep and goats, similar to the effects in cattle, which causes major economic damage. Regular vaccinations alongside good...
BACKGROUND AND AIM
causes respiratory infection and mortality in sheep and goats, similar to the effects in cattle, which causes major economic damage. Regular vaccinations alongside good management practices remain the most efficient tools for controlling this disease. Indeed, vaccines against pasteurellosis are available, but results on their efficacy have varied. Therefore, this study aimed to evaluate the efficacy of three vaccines against mannheimiosis in small ruminants.
MATERIALS AND METHODS
We evaluated three vaccines developed from a local field isolate based on the inactivated bacterium, its toxoid, and a mixture of bacterin/toxoid, which we then tested on sheep and goats. Selected criteria that were evaluated were safety, antibody response, and protection through a challenge. Post-vaccination monitoring was carried out by enzyme-linked immunosorbent assay. The evaluation was based on antibody responses to vaccination in sheep and goats for both bacteria and leukotoxin. Protection was assessed by clinical and lesion scores after the challenge of vaccinated goats with a pathogenic strain.
RESULTS
The three tested vaccines were completely safe, did not cause any adverse reactions, and induced significant antibody titers in immunized animals. Following challenge, unvaccinated goats showed clinical signs with lesions typical of the disease. Meanwhile, the best protection was obtained with the inactivated combined bacterin/toxoid vaccine.
CONCLUSION
This study highlighted the effectiveness of adding a bacterial toxoid in the vaccine as a promising solution for preventing mannheimiosis in small ruminants. Because of the worldwide distribution of infection, general prophylaxis based on a combined inactivated vaccine could greatly benefit.
PubMed: 36855364
DOI: 10.14202/vetworld.2023.68-75 -
Veterinary Microbiology May 2023A hierarchical cluster analysis was used to classify outbreaks of bovine respiratory disease (BRD; n = 156) in natural groups according to the detection of nine...
A hierarchical cluster analysis was used to classify outbreaks of bovine respiratory disease (BRD; n = 156) in natural groups according to the detection of nine pathogens (parainfluenza 3 virus (PI-3), bovine respiratory syncytial virus (BRSV), bovine coronavirus (BCV), bovine viral diarrhea virus (BVDV), and bovine herpesvirus 1 (BHV-1), Mannheimia haemolytica, Pasteurella multocida, Histophilus somni, and Mycoplasma bovis. Pathogens were detected by individual q-PCRs. Two clusters were identified. Cluster 1 was characterized by a relatively high frequency (40-72%) of four BRD-associated viruses, supporting their primary involvement in BRD. Cluster 2 was characterized by frequencies of PI-3, BRSV, or BVDV below 10% each. P. multocida and M. haemolytica were detected with high frequencies in both clusters (P > 0.05), while M. bovis and H. somni showed a significantly higher frequency in cluster 1and 2, respectively. Outbreaks in cluster 1 were associated with preweaning calves younger than 5 months (OR 2.2; 95% CI 1.1-4.5) and with cold months, whereas cluster 2 was associated with fattening calves older than 5 months after arrival to feedlots and without any seasonality. Thus, in addition to the classic epidemiological BRD pattern characterized by the primary involvement of viruses occurring preferably during winter and affecting young calves, there is a second pattern in which viruses would be less relevant, affecting mainly calves older than 5 months at any time of the year. This study allows a better understanding of the BRD epidemiology, which can be useful when implementing management and prophylaxis measures for a better control of this disease.
Topics: Animals; Cattle; Cattle Diseases; Respiratory Tract Diseases; Mannheimia haemolytica; Pasteurella multocida; Diarrhea Viruses, Bovine Viral; Disease Outbreaks; Cluster Analysis
PubMed: 36848816
DOI: 10.1016/j.vetmic.2023.109701 -
Antibiotics (Basel, Switzerland) Jan 2023Antimicrobial resistance (AMR) is a global public health threat that jeopardizes efficacy of antibiotics in veterinary and human medicine. Antibiotics are commonly...
Antimicrobial resistance (AMR) is a global public health threat that jeopardizes efficacy of antibiotics in veterinary and human medicine. Antibiotics are commonly administered to target the bacterial component of bovine respiratory disease (BRD). The objectives of this study were to obtain a better understanding of antibiotic resistance in BRD-associated bacteria (, and ), investigate the clinical significance of AMR by monitoring clinical outcomes, and determine if regional differences exist in AMR trends. Deep pharyngeal swabs were used to sample beef cattle at initial BRD diagnosis ( = 453) from US feedlots representing three geographic regions. Organisms were identified by bacterial culture and subjected to broth microdilution antimicrobial susceptibility testing. Bacterium prevalence include (36.0%), (32.7%) and (28.5%). Of the isolates, 39.5% were resistant to at least one antimicrobial, compared to 11.7% and 8.8% and , respectively. Non-susceptibility across all organisms was 5.7 X more likely in animals that received metaphylaxis, than those that did not ( < 0.0001; OR 5.7; CI 2.6-12.5). During days on feed 21-40, non-susceptibility of was 8.7 X more likely than ( = 0.0002; OR 8.7; CI 2.8 to 27.4) and 6 X more likely than ( = 0.0016; OR 6.0; CI 2.0-18.0).
PubMed: 36830126
DOI: 10.3390/antibiotics12020215 -
Journal of Dairy Science Apr 2023The aim of this study was to evaluate the effect of therapeutically administered tildipirosin or florfenicol + flunixin meglumine for the treatment of bovine respiratory...
Treatment with 2 commercial antibiotics reduced clinical and systemic signs of pneumonia and the abundance of pathogenic bacteria in the upper respiratory tract of preweaning dairy calves.
The aim of this study was to evaluate the effect of therapeutically administered tildipirosin or florfenicol + flunixin meglumine for the treatment of bovine respiratory disease (BRD) accompanied by fever in calves before weaning compared with diseased and untreated animals. As specific objectives, we evaluated the composition of the bacterial microbiota of the upper respiratory tract (URT) and blood and health parameters of the animals. Preweaning Holstein female calves diagnosed with naturally acquired pneumonia were randomly assigned to one of the following experimental groups on the day of diagnosis (d 0): (1) TLD (n = 36): single subcutaneous injection with 4 mg/kg tildipirosin; (2) FLF (n = 33): single subcutaneous injection with an antimicrobial (40 mg/kg florfenicol) combined with a nonsteroidal anti-inflammatory drug (2.2 mg/kg flunixin meglumine); and (3) NEG (n = 35): no treatment within the first 5 d following enrollment. The NEG treatment group was closely monitored for 5 d, and calves were removed from the study following a standardized late treatment protocol, when necessary, to minimize health concerns. Healthy untreated calves (CTR; n = 31) were also selected for the study and used as controls. Blood samples used for biochemical analysis and nasopharyngeal swabs used for evaluation of URT microbiota were collected daily from d 0 until d 5 and then weekly until weaning. Next-generation sequencing of the 16S rRNA gene was used to assess the URT microbiota at the phylum and genus levels. Clinical signs associated with pneumonia and otitis media were assessed daily, as was the need for antibiotic interventions. Calves in the TLD and FLF groups had faster recovery from fever within the first 5 d after enrollment. In addition, antibiotic-treated calves reached the same serum haptoglobin levels as healthy calves on d 2 after diagnosis, whereas calves in the NEG group had higher haptoglobin levels than the CTR group until at least d 5 after BRD diagnosis. Calves in the TLD and FLF groups had a lower risk of treatment for pneumonia (FLF = 22.8%; TLD = 27.7%) from d 5 to weaning than calves in the NEG group (54.7%). Furthermore, FLF treatment had a significantly lower risk of nasal discharge, otitis media, and treatment failure compared with the NEG group, but did not differ from the TLD group. Differences in the composition of the URT microbiota were found between groups, and the genus Mycoplasma was the most abundant in samples collected from the URT of calves with and without pneumonia. Both drugs were effective in reducing the mean relative abundance (MRA) of important genera associated with pneumonia (Mannheimia and Pasteurella), although an increase in Mycoplasma MRA was observed for tildipirosin-treated calves. In conclusion, both drugs were effective in reducing the inflammatory signs of pneumonia and the need for antimicrobial treatment after enrollment compared with no treatment. In addition, both TLD and FLF were effective in reducing the MRA of important bacterial genera associated with pneumonia; however, TLD treatment was associated with increased Mycoplasma MRA compared with healthy and untreated calves.
Topics: Animals; Cattle; Female; Anti-Bacterial Agents; RNA, Ribosomal, 16S; Haptoglobins; Bacteria; Pneumonia; Cattle Diseases; Otitis Media; Respiratory System
PubMed: 36797182
DOI: 10.3168/jds.2022-22451 -
Scientific Reports Feb 2023Each year, bovine respiratory disease (BRD) results in significant economic loss in the cattle sector, and novel metabolic profiling for early diagnosis represents a...
Each year, bovine respiratory disease (BRD) results in significant economic loss in the cattle sector, and novel metabolic profiling for early diagnosis represents a promising tool for developing effective measures for disease management. Here, H-nuclear magnetic resonance (H-NMR) spectra were used to characterize metabolites from blood plasma collected from male dairy calves (n = 10) intentionally infected with two of the main BRD causal agents, bovine respiratory syncytial virus (BRSV) and Mannheimia haemolytica (MH), to generate a well-defined metabolomic profile under controlled conditions. In response to infection, 46 metabolites (BRSV = 32, MH = 33) changed in concentration compared to the uninfected state. Fuel substrates and products exhibited a particularly strong effect, reflecting imbalances that occur during the immune response. Furthermore, H-NMR spectra from samples from the uninfected and infected stages were discriminated with an accuracy, sensitivity, and specificity ≥ 95% using chemometrics to model the changes associated with disease, suggesting that metabolic profiles can be used for further development, understanding, and validation of novel diagnostic tools.
Topics: Animals; Cattle; Male; Cattle Diseases; Respiratory Tract Diseases; Respiration Disorders; Mannheimia haemolytica; Magnetic Resonance Spectroscopy; Metabolomics; Plasma; Respiratory Syncytial Virus Infections
PubMed: 36792613
DOI: 10.1038/s41598-023-29234-3