Contact: Istituto Zooprofilattico Sperimentale dell’Abruzzo e del Molise “G. Caporale” brucellosis2022.izs.it brucellosis2022@izs.it
P8-03 16S Microbiome Characterization of Brucella Positive and Negative Raw Milk Samples Collected from three regions in Tanzania.
2022, 8 - Epidemiology, control and eradication
2022

P8-03 16S Microbiome Characterization of Brucella Positive and Negative Raw Milk Samples Collected from three regions in Tanzania.

8 - Epidemiology, control and eradication
Published 2023-02-10
Joram Buza
Nelson Mandela African Institution of Science and Technology, Arusha, Tanzania
Zachariah Ephraim Makondo
Tanzania Veterinary Laboratory Agency, Dar es Salaam, Tanzania
Happiness Kumburu
Kilimanjaro Clinical Research Institute, Moshi, Tanzania
Beatus Lyimo
Nelson Mandela African Institution of Science and Technology, Arusha, Tanzania
Ray Kayaga
Tanzania Veterinary Laboratory Agency, Dar es Salaam, Tanzania
Charles Mayenga
Tanzania Veterinary Laboratory Agency, Dar es Salaam, Tanzania
Earl Austin Middlebrook
Los Alamos National Lab, Los Alamos, NM, U.S.A
Erika Ganda
Department of Animal Science, Pennsylvania State University, University Park, PA, U.S.A
Nammalwar Sriranganathan
Biomedical Sciences and Pathobiology, Virginia Tech, Blacksburg, VA 24061. USA
Jeanne Marie Fair
Los Alamos National Lab, Los Alamos, NM, U.S.A
Andrew William Bartlow
Los Alamos National Lab, Los Alamos, NM, U.S.A
Blandina Mmbaga
Kilimanjaro Clinical Research Institute, Moshi, Tanzania
Vivek Kapur
Nelson Mandela African Institution of Science and Technology, Arusha, Tanzania; Department of Animal Science, Pennsylvania State University, University Park, PA, U.S.A; Huck Institutes of Life Sciences, Pennsylvania State University, University Park, PA, U.S.A
Robab Katani
Nelson Mandela African Institution of Science and Technology, Arusha, Tanzania; Huck Institutes of Life Sciences, Pennsylvania State University, University Park, PA, U.S.A

Keywords

16S Microbiome
Tanzania
Raw milk
Brucella Positive and Negative
Cattle

Categories

Abstract

Brucellosis is endemic in many low- and middle-income countries, including Tanzania, where the disease is often transmitted via the consumption of raw milk or direct contact with infected animals/products. Brucella species can localize in the mammary lymph nodes and glands of infected dairy animals that may shed the pathogen in milk for extended periods of time and present a significant health risk to consumers of unpasteurized dairy products and individuals in direct contact with infected animals. Since the sale and consumption of raw and unprocessed milk is common amongst pastoral and agropastoral communities in Tanzania, large proportions of the population are susceptible to milk-borne and zoonotic diseases, including brucellosis. We have recently initiated a five-year cross-sectional survey to assess risk factors associated with brucellosis, including raw milk consumption in Tanzania. To determine the total microbial diversity present inraw milk and to identify potential associations with other pathogens or microbial signatures of milk from infected animals, characterization of Brucella-positive and negative samples using next-generation sequencing is being performed. In brief, we have collected 176 bovine milk samples from dairy cattle (76), and Boran crossbred cattle (100) from seven herds in three regions of Tanzania, Dodoma (Central), Morogoro (East), and Tanga (Northeast). Milk samples were collected aseptically, and Fluorescence Polarization Assay (FPA) was used to screen the milk samples as an initial screen to ascertain Brucella’s status. DNA extraction from the FPA+ and select FPA- milk samples were performed by applying a magnetic-based extraction method, followed by real-time PCR-based detection of the IS711 insertion sequence to confirm the Brucella status. Studies to assess 16S rRNA-based microbiome diversity will next be performed, and genomic DNA will undergo PCR with primers 515F-806R to amplify the V4 region of the 16S gene for microbiome profiling following the Earth Microbiome Project protocol Next-generation sequencing of the 16S RNA and Illumina short-read sequencing and are in progress.