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Bovine rhinitis virus (BRV) is an important pathogen responsible for the bovine respiratory disease complex (BRDC) and can be divided into two genotypes (BRAV and BRBV). To establish a duplex quantitative real-time RT-PCR assay for simultaneous detection of BRAV and BRBV, specific primers and TaqMan probes targeting the 5'NTR of BRAV and 3'NTR of BRBV were designed. A duplex quantitative real- time RT- PCR assay for simultaneous detecting BRAV and BRBV was preliminarily established by optimizing reaction conditions for each step. The assay specifically detects BRAV and BRBV, and no crossreaction with other common bovine respiratory pathogens, including IDV, BCoV, BVDV-1, BRSV, BPIV-3, BAdV-3, mycoplasma bovis, Pasteurella multocida, Mannheimia haemolytica, Escherichia coli, and Salmonella, was observed. In addition, the sensitivity test showed that the detection limits of this assay were 3.2x101 copies/L for both BRAV and BRBV plasmid standards. Besides, the repeatability test showed that the variation coefficients of this assay were less than 0.05 from both lot-to-lot and intra-lot. These results showed that the assay has high specificity, extreme sensitivity, and good repeatability. Moreover, a total of 43 nasal swabs of BRDC cattle were tested by our assay and four other quantitative real-time RT-PCR assays, including 3 BRAV assays and 4 BRBV assays. The results showed that the detection rates of our assay were 32.56%(14/43) for BRAV and 30.23%(13/43) for BRBV, and the detection rates of other quantitative real-time RT-PCR assays were 0(0/43), 2.33%(1/43), 23.26%(10/43) for BRAV and 27.91% (12/43), 27.91%(12/43), 27.91%(12/43), 27.91%(12/43) for BRBV, indicating that our assay has a more substantial detection capability than other assays. This study firstly established a duplex quantitative real-time RT-PCR assay for simultaneous detection of BRAV and BRBV, and the assay exhibited high specificity, sensitivity, and stability. Moreover, the study firstly confirmed the existence of BRAV in China, contributing to the prevention and control of BRDC.
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Objective: The aim of this study was to establish a one-step multiplex real-time RT-PCR method to simultaneously detect and quantify five swine diarrhea related viruses, PEDV, GARV, PDCoV, SADS-CoV and PTV, so as to provide an efficient and sensitive tool for rapid diagnosis and epidemiological investigation of porcine diarrhea. Method: The ORF3 gene sequences of several genotypes of PEDV were analyzed, and then the primers and probes were designed for detection of PEDV field strains by referring to the ORF3 genes, which contained deletion mutations in attenuated strains. The 5'-end conserved region of NSP5 genes of GARV G3, G4, G5 and G9 strains were analyzed for design of probes and primers. The specific primers and probes targeting to the conserved regions of PDCoV M, PTV 5'UTR and SADS-CoV N genes were designed for detection of the pathogens. The ROC curves were completed by referring to parameters that were set in RStudio. The specificity value, sensitivity value, and areas under the curves (AUC) and Youden value were calculated according to ROC curves to determine the cut-off CT value. The amplified fragments were cloned into pEASY-T1 vector. The standards prepared through in vitro transcription were named as cRNA-PEDV, cRNA-GARV, cRNA-PDCoV, cRNA-PTV and cRNA-SADS-CoV. The sensitivity, specificity and repeatability of one-step multiplex real-time RT-PCR were evaluated. Coincidence rate between this and another similar method were compared in the detection of clinical samples. Result: Both the annealing temperature and optimal concentrations of primers and probes were obtained for detection of the five pathogens. According to the ROC curve, the CT cut off values for detection of PEDV, GARV, PDCoV, PTV, and SADS-CoV were set as 35.78, 34.25, 34.98, 34.60, and 35.70, respectively. The detection sensitivity of this method for the five pathogens could reach 1..102 copies/L. The standard curves had a good linear relationship and the amplification efficiency was between 96.3% and 104%. The established method could not detect the PEDV vaccine strains and other swine infecting viruses and bacteria including TGEV, CSFV, PRV, PRRSV, S.choleraesuis, P.multocida, E.coli, S.suis and S.aureus. The repeatability test showed the range of intra-assay and inter-assay coefficients of variability: 0.22% to 3.08% and 0.89% to 4.0%, respectively. The detection coincidence rates of the established detection method and another similar method for the five pathogens in 242 clinical samples were 97.9%, 98.8%, 100%, 98.3% and 100% for PEDV, GARV, PDCoV, PTV and SADS-CoV, respectively. The Kappa values were all higher than 0.9. The method had advantage over a commercial diagnostic kit for detection of PEDV wild strains in accuracy. Detection results with clinical samples showed that positive rates of PEDV, GARV, PDCoV and PTV was 10.7% (26/242), 13.6% (33/242), 18.2% (44/242) and 14.5% (35/242), respectively, demonstrating the prevalence state of the four pathogens in Sichuan province in the years. SADS-CoV was not detectable in any areas, but the phenomenon of coinfection with different diarrhea causing viruses was common. Therefore, it was necessary to strengthen the surveillance of several porcine diarrhea viruses in Sichuan province for preventive control. Conclusion: In this study, a one-step multiplex real-time RT-PCR was established for simultaneous detection of PEDV wild strains, PDCoV, SADS-COV and GARV, PTV multiple genotypes, which provided an efficient and sensitive tool for the differential diagnosis and epidemiological investigation of swine diarrhea disease.
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OBJECTIVES: In COVID-19 patients, bacterial and fungal pulmonary coinfections, such as Streptococcus pneumoniae, Staphylococcus aureus, Haemophilus influenzae, or Aspergillus, have been reported, but to our knowledge, no case has been reported due to Pasteurella multocida. PATIENTS AND METHODS: We describe three cases of Pasteurella multocida coinfections occurring during the 4th wave of COVID-19 in Martinique (French West Indies). RESULTS: All three cases were fatal; thus, Pasteurella multocida has to be considered as a potentially severe coinfection agent. CONCLUSIONS: Alteration of the epithelial-endothelial barrier due to a SARS-CoV-2 infection probably promotes the expression of a Pasteurella infection. In addition, the SARS-CoV-2 infection induced immunosuppression, and an inflammatory cascade could explain the infection's severity. The use of corticosteroids, which are part of the first-line therapeutic arsenal against COVID-19, may also promote the pathogenicity of this agent.
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SESSION TITLE: Atypical Cases of Sepsis SESSION TYPE: Rapid Fire Case Reports PRESENTED ON: 10/19/2022 12:45 pm - 1:45 pm INTRODUCTION: Pasteurella multocida is a Gram-negative coccobacillus that causes infections after animal bites or scratches. It typically manifests as cellulitis but severe infections are possible though rare. We present a case of an immunocompetent man with COVID-19 who developed septic shock due to P. multocida bacteremia and pneumonia with no evidence of wound infection. CASE PRESENTATION: A 59-year-old Hispanic man with a history of anxiety and HLD presented with 10 days of nausea, vomiting, chills, and nonproductive cough. He was initially afebrile, on room air but tachycardic. His physical exam was unremarkable. Labs revealed WBC 10x10*3/uL, procal 3.3 ng/mL, negative lactic acid, and positive COVID-19. CT chest showed a right upper lobe consolidation with bilateral patchy infiltrates. He was admitted for sepsis secondary to COVID and superimposed bacterial pneumonia. Ceftriaxone, azithromycin, remdesivir, and dexamethasone were started. Overnight, the patient desaturated to low 80s and required HFNC FiO2 65%. In the morning, FiO2 increased to 80%. ICU was called and upon their assessment, the patient was febrile, tachycardic, tachypneic, hypotensive, and saturating 87-88% on HFNC FiO2 70%. Labs showed WBC 3.1 with left shift, Cr 1.7 mg/dL, lactic acid 5 mmol/L, and procalcitonin >100. He was intubated given persistent hypoxia and increased work of breathing. Antibiotics were broadened to vancomycin, pip/tazo, and azithromycin. The patient acutely decompensated after intubation, requiring multiple high-dose pressor support. Prelim blood cultures grew Gram-negative bacteria so antibiotics were broadened to meropenem. TTE was negative for endocarditis. Pressors were eventually weaned and the patient was extubated. Blood cultures grew P. multocida in 4/4 bottles so meropenem was narrowed to penicillin. His family reported that he was living at a friend's house with cats around but was unaware of any bites or scratches and he had no history of splenectomy. No portal of entry was noted upon careful skin examination. The patient continued to improve clinically with procal that rapidly downtrended. He was eventually discharged home. DISCUSSION: The mortality for severe P. multocida presentations is about 25-30%. Severe cases are generally reported in elderly, immunocompromised, or young immunosuppressed patients. We report what is to our knowledge, the first case of a severe P. multocida infection in an immunocompetent middle-aged man in the background of a COVID-19 infection. It is unclear the degree of COVID contribution and if his bacteremia preceded the pneumonia. His morbidity was primarily driven by the P. multocida bacteremia and pneumonia given the localized right upper lobe consolidation, elevated procal that rapidly decreased with antibiotics, and quick improvement and extubation. CONCLUSIONS: P. multocida infection should be considered in any patient with septic shock and exposure to animals. Reference #1: Blain H, George M, Jeandel C. Exposure to domestic cats or dogs: risk factor for Pasteurella multocida pneumonia in older people? Journal of the American Geriatrics Society. 1998;46(10):1329-1330. Reference #2: Tseng HK, Su SC, Liu CP, Lee CM. Pasteurella multocida bacteremia due to non-bite animal exposure in cirrhotic patients: report of two cases. Journal of microbiology, immunology, and infection= Wei mian yu gan ran za zhi. 2001;34(4):293-296. Reference #3: Kofteridis DP, Christofaki M, Mantadakis E, et al. Bacteremic community-acquired pneumonia due to Pasteurella multocida. International Journal of Infectious Diseases. 2009;13(3):e81-e83. doi:10.1016/j.ijid.2008.06.023 DISCLOSURES: No relevant relationships by Joanne Lin No relevant relationships by Harjeet Singh No relevant relationships by Jose Vempilly No relevant relationships by Joshua Wilkinson
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Broiler population is one of the most important segments of livestock due to its significant contribution in white meat production. Infectious disease outbreaks adversely influence the production potential and consequently cause economic losses. Epidemiological data regarding magnitude of these disease outbreaks is of fundamental importance for planning of a comprehensive control strategy. With retrospective design, this study was conducted from January 2013 through December 2017 in order to assess the disease burden on broilers reared in different open type poultry houses. Out of total 658 commercial farms with capacity of 4221800 broilers, across Chakwal, a representative sample of 70 farms with capacity of 448000 broilers was randomly selected for collection and analysis of disease data. Five years' data of these randomly selected farms revealed highest (44.64%) crude morbidity during monsoon season followed by 23.92%, 22.12% and 17.49% for winter, spring and post-monsoon seasons respectively. The highest (14.90%) prevalence was recorded for new castle disease followed by infectious bursal disease (11.79%), pullorum disease (11.17%), colibacillosis (8.71%), infectious bronchitis (7.87%), inclusion body hepatitis (7.79%), chronic respiratory disease (7.67%), necrotic enteritis (6.48%), coccidiosis (6.09%), mycotoxicosis (5.43%), fowl cholera (4.74%), infectious coryza (4.41%), fowl typhoid (4.22%), omphalitis (3.71%) and hydropericardium syndrome (0.05%). Maximum share in crude morbidity was contributed by bacterial diseases with highest proportional morbidity of 48.68% followed by viral (40.32%), parasitic (5.80%) and fungal (5.20%) diseases. This epidemiological data represents true picture of study population and is a valuable tool for planning of prevention strategy and research priorities.
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Bovine respiratory disease (BRD) is considered a major cause of morbidity and mortality in young calves and is caused by a range of infectious agents, including viruses and bacteria. This study aimed to determine the frequency of viral and bacterial pathogens detected in calves with BRD from high-production dairy cattle herds and to perform the molecular characterization of N and S1 genes in identified bovine coronavirus (BCoV) strains. Nasal swabs were collected from 166 heifer calves, namely, 85 symptomatic and 81 asymptomatic calves aged between 5 and 90 days, from 10 dairy cattle herds. Nasal swabs were evaluated using molecular techniques for the identification of viruses (BCoV, bovine alphaherpesvirus 1, bovine viral diarrhea virus, bovine parainfluenza virus 3, and bovine respiratory syncytial virus) and bacteria (Pasteurella multocida, Mannheimia haemolytica, Histophilus somni, and Mycoplasma bovis). In addition, five and two BCoV-positive samples were submitted to N and S1 gene amplification and nucleotide sequencing, respectively. The frequency of diagnosis of BCoV was higher (56%, 93/166) than the frequency of P. multocida (39.8%, 66/166) and M. haemolytica (33.1%, 55/166). The three microorganisms were identified in the calves of symptomatic and asymptomatic heifer calve groups. All other pathogens included in the analyses were negative. In the phylogenetic analysis of the S1 gene, the Brazilian strains formed a new branch, suggesting a new genotype, called # 15; from the N gene, the strains identified here belonged to cluster II. This study describes high rates of BCoV, P. multocida, and M. haemolytica in heifer calves from high-production dairy cattle herds with BRD. Additionally, the molecular characterization provides evidence that the circulating BCoV strains are ancestrally different from the prototype vaccine strains and even different BCoV strains previously described in Brazil.
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The number of pets such as dogs, cats, rabbits, and parrots has increased in European families. Social benefits to owners such as decreasing feelings of loneliness and anxiety are provided by pets which are also used in Animal-assisted Therapy (AAT). Nevertheless, human-animal interactions are also associated with health problems including allergies, asthma, and zoonosis. Rabbits may carry potential pathogens for humans. One of the most common bacteria that colonizes the oro-pharynx and the upper respiratory tract of rabbits is Pasteurella (P.) multocida. Transmission of the infection to humans results from scratches, licks, and bites but it also can occur from the inhalation of air particles containing the microorganism. Immunocompromised people or persons with pulmonary disorders are particularly susceptible to the infection. Infected rabbits may carry P. multocida with or without clinical signs. In this paper, the sensitivity to antibiotics and the invasiveness ability of P. multocida identified in a farm of pet rabbits affected by severe pasteurellosis were investigated. The strain was P. multocida belonging to capsular type A which is the type most often detected in humans. The identified strain was susceptible to the tested antibiotics, but it appeared equipped with several virulence genes which are responsible for fimbriae production, adhesion processes to host cells, enzyme production, and are involved in iron acquisition processes. These findings are of particular interest because rabbits recovered from pasteurellosis very often become carriers of the bacteria. Therefore, we suggest considering P. multocida screening in the routine medical checks of rabbits, especially if they are meant to be companion animals for children and elder people, given that the transmission of the pathogen cannot be excluded.
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Today, pets are the source of numerous infectious diseases that can be transmitted to humans, as a result of their increasingly frequent contact. The most important viruses with zoonotic potential include rabies and influenza viruses as well as rotaviruses and noroviruses. However, the importance of individual viruses varies depending on the climate and infectious disease control systems in certain countries. Dogs, cats, and other increasingly popular types of pets can transmit bacterial zoonotic agents to humans in various ways. In addition to known pathogens such as the bacteria causing leptospirosis, salmonellosis, campylobacteriosis, or brucellosis, the bacteria Pasteurella multocida and Bartonella henselae transmitted by bites or scratches are also significant in human pathology. There has been a significant increase in the prevalence of methicillin-resistant strains of Staphylococcus aureus in isolates originating from pets and the transmission of these strains between humans and animals requires special attention. Furthermore, fungi causing diseases such as sporotrichosis or dermatophytosis are linked to long-term and persistent infections in humans. The epidemiological situation caused by SARS-CoV-2, and the assumption of an interspecies jump of this virus from animals to humans, including its documented presence in domestic cats, dogs, tigers, and martens, have raised the question of the possibility of virus transmission from pets to humans. However, the current pandemic is caused exclusively by SARS-CoV-2 transmission in the human population, and these animals are not a source of infection for humans. A significant number of zoonoses originating from pets is a threat to public health, thus requiring the "One Health" approach through close cooperation between human and veterinary medicine to develop and implement effective health measures for both humans and animals. As part of responsible ownership, pet owners must be informed by veterinarians about measures to prevent infectious diseases and certain risks that are related to keeping certain species of animals.