Phagocytosis and intracellular killing of Pasteurella multocida B:2 by macrophages: A comparative study between buffalo and cattle

Hasnan, Qistina; Puspitasari, Yulianna; Othman, Siti Sarah; Zamri‐Saad, Mohd; Salleh, Annas

doi:10.14202/vetworld.2022.275-280

Cited by 3 publications

(2 citation statements)

References 26 publications

(29 reference statements)

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“…In addition, Pomeroy and collaborators found a significant association between cattle postpartum diseases (mastitis and metritis) and modifications of monocyte subset counts (Pomeroy et al., 2017). Monocytes isolated from cattle and buffalo blood and differentiated into macrophages showed differential phagocytosis and intracellular killing abilities against the bacterial pathogen ( Pasteurella multocida ) with higher efficiency for cattle than buffalo cells (Hasnan, Puspitasari, Othman, Zamri‐Saad, & Salleh, 2022).…”

Section: Introductionmentioning

confidence: 99%

Flow Cytometric Identification and Enumeration of Monocyte Subsets in Bovine and Water Buffalo Peripheral Blood

et al. 2023

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Monocytes are innate immune system key players with pivotal roles during infection and inflammation. They migrate into tissues and differentiate into myeloid effect cells (macrophages, dendritic cells) which orchestrate inflammatory processes and are interfaces between the innate and adaptive immune responses. Their clinical relevance to health and disease of cattle (Bos taurus) and water buffalo (Bubalus bubalis), two of the most important livestock species, has been highlighted in physiologic (pregnancy) and pathologic (mastitis, metritis, and viral infections) conditions. The existence of three different monocyte subsets in cattle was established by flow cytometry (FC), as follows: classical (cM; CD14++CD16–/low), intermediate (intM; CD14++/+CD16+), and non‐classical (ncM; CD14–/lowCD16++) monocytes. FC applications for studying the immune system of cattle and water buffalo still have significant limitations. In this article, we describe some practical approaches to overcome these limitations and, in particular, allow the identification and enumeration of cM, intM, and ncM subpopulations in cattle and buffalo peripheral blood. Indeed, we propose the new procedure lyse/wash/no‐centrifugation (L/W/NC) that can be combined with the FC absolute counting procedures and can overcome specific issues of the lyse/no‐wash protocols (L/NW). Finally, for the first time, we demonstrated the existence of cM, intM, and ncM monocyte subsets also in the water buffalo, showing some interesting differences with cattle, such as the bubaline cM are mainly CD14+/++/CD16+. These subtle differences may influence inflammatory disease regulation in, for example, mastitis and metritis. The upregulation of CD16 expression on cM may reveal different monocyte priming, leading to different functional features of macrophages/dendritic cells in tissues after infection. © 2023 Wiley Periodicals LLC. Basic Protocol: Absolute count of cM, intM, and ncM without compensation Alternate Protocol: Absolute count of cM, intM, and ncM for single laser platform Support Protocol 1: In‐house monoclonal antibody labeling using a Pacific Blue™ kit Support Protocol 2: In‐house monoclonal antibody labeling using an Alexa Fluor® 647 kit Support Protocol 3: Titration of fluorochrome‐conjugated antibodies

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Section: Introductionmentioning

confidence: 99%

Flow Cytometric Identification and Enumeration of Monocyte Subsets in Bovine and Water Buffalo Peripheral Blood

et al. 2023

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“…The course of BRD can range from 3 days to 1 week with respiratory symptoms such as dry cough, dyspnea, and fibrinous types of pneumonia ( 12 ). HS is caused by serogroups B and E of Pm ( 19 , 20 ). HS shows high mortality for ill cattle with acute symptoms of high fever, diarrhea, and death within 12–24 h ( 12 ).…”

Section: Introductionmentioning

confidence: 99%

Construction of a one-step multiplex real-time PCR assay for the detection of serogroups A, B, and E of Pasteurella multocida associated with bovine pasteurellosis

et al. 2023

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Bovine pasteurellosis, caused by serogroups A, B, and E of Pasteurella multocida (Pm), is mainly manifested as bovine respiratory disease (BRD) and hemorrhagic septicemia (HS). The disease has caused a great economic loss for the cattle industry globally. Therefore, identifying the Pm serogroups is critical for optimal diagnosis and subsequent clinical treatment and even epidemiological studies. In this study, a one-step multiplex real-time PCR assay was established. Three pairs of specific primers were prepared to detect the highly conserved genomic regions of serogroups A (HyaD), B (bcbD), and E (ecbJ) of Pm, respectively. The results depicted that the method had no cross-reaction with other bovine pathogens (Mannheimia hemolytica, Escherichia coli, Listeria monocytogenes, Staphylococcus aureus, Salmonella Dublin, Mycobacterium paratuberculosis, infectious bovine rhinotracheitis virus, and Mycoplasma bovis). The linear range (107 to 102 copies/μL) showed the R2 values for serogroups A, B, and E of Pm as 0.9975, 0.9964, and 0.996, respectively. The multiplex real-time PCR efficiency was 90.30%, 90.72%, and 90.57% for CartA, CartB, and CartE, respectively. The sensitivity result showed that the serogroups A, B, and E of Pm could be detected to be as low as 10 copies/μL. The repeatability result clarified that an intra-assay and an inter-assay coefficient of variation of serogroups A, B, and E of Pm was < 2%. For the clinical samples, the detection rate was higher than the OIE-recommended ordinary PCR. Overall, the established one-step multiplex real-time PCR assay may be a valuable tool for the rapid and early detection of the serogroups A, B, and E of Pm with high specificity and sensitivity.

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Sero-Prevalence of Hemorrhagic Septicaemia in Cattle and Buffalo Population of Indian States Karnataka and Gujarat

Shome,

Kanani,

Gurrappanaidu

et al. 2024

Veterinary Sciences

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Hemorrhagic septicemia (HS) is a highly contagious and fatal disease of cattle and buffaloes caused by P. multocida. Both conventional and molecular methods are applied in parallel for rapid diagnosis of HS outbreaks and the periodical surveillance strategy to identify risk areas for HS is ignored. The current cross-sectional study aimed to estimate sero-prevalence and associated risk factors for HS in cattle and buffaloes in non-vaccinated regions of two Indian states. HS surveillance was carried out through the multi-stage random sampling technique at different strata. The study employed a questionnaire incorporating host factors (species, breed, sex, age, and lactation) and demographic parameters (state, district, block/cluster and village/epiunits, and household). First, two Indian states known for high milk production were selected followed by two districts within each state, subsequently four clusters within each district, finally 5–10 epiunits within clusters and 5–8 households within clusters were randomly selected to collect cattle and buffalo samples. The chi-square/p values and maps were prepared to represent disease prevalence and to correlate disease risk factors at different strata. A total of 692 cattle and buffalo serum samples were sourced from two states of the country (Karnataka-285 and Gujarat-407). In the first strata, antibodies to P. multocida were high in Gujarat (14.49%, CI: 11.22–18.30) compared to Karnataka (3.85%, CI: 1.94–6.80) with significant (p < 0.0001) association between the states. In the second strata, one of the four districts investigated revealed the highest sero-prevalence (18.61%, CI: 13.81–24.24) with statistical significance (p = 0.01) between the districts. Among clusters, one out of eight clusters showed the highest sero-prevalence (23.02%, CI: 16.59–30.54) with statistical significance (p = 0.03) between the clusters in the third strata. At epiunit level (fourth strata), 9 out of 27 epiunits (33.33%) visited in Karnataka and 24 out of 29 epiunits sampled in Gujarat were sero-positive (82.75%) in iELISA. At the household level, out of 306 HH visited, 40 HH had at least one positive animal (13.07%) and the p value between HH in the two states was highly significant (p = 0.0002). Chi-square analysis did not find any association of HS sero-prevalence to species, age, and lactation. However, significantly higher (p < 0.05) sero-prevalence was recorded in indigenous cattle breeds (16.56%) compared to crossbreeds (6.59%). Various immunoprophylactics and antibiotic therapies are effective against HS, but inappropriate disease reporting and failure to implement adequate vaccination control measures are the gaps identified. The present study highlights the current scenario of HS sero-prevalence in two of the high milk-producing states of India, which will be useful for stakeholders for undertaking the implementation of surveillance and control strategies for the regions.

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Phagocytosis and intracellular killing of Pasteurella multocida B:2 by macrophages: A comparative study between buffalo and cattle

Cited by 3 publications

References 26 publications

Flow Cytometric Identification and Enumeration of Monocyte Subsets in Bovine and Water Buffalo Peripheral Blood

Flow Cytometric Identification and Enumeration of Monocyte Subsets in Bovine and Water Buffalo Peripheral Blood

Construction of a one-step multiplex real-time PCR assay for the detection of serogroups A, B, and E of Pasteurella multocida associated with bovine pasteurellosis

Sero-Prevalence of Hemorrhagic Septicaemia in Cattle and Buffalo Population of Indian States Karnataka and Gujarat

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