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20 pages, 1874 KiB

Open AccessArticle

Bayesian Estimation of the Prevalence and Test Characteristics (Sensitivity and Specificity) of Two Serological Tests (RB and SAT-EDTA) for the Diagnosis of Bovine Brucellosis in Small and Medium Cattle Holders in Ecuador

by Valeria Paucar, Jorge Ron-Román, Washington Benítez-Ortiz, Maritza Celi, Dirk Berkvens, Claude Saegerman and Lenin Ron-Garrido

Microorganisms 2021, 9(9), 1815; https://doi.org/10.3390/microorganisms9091815 - 26 Aug 2021

Cited by 5 | Viewed by 2675

Abstract

In Ecuador, a national program for bovine brucellosis control has been in implementation since 2008. Given the costs, small- and medium-sized livestock holders are not completely committed to it. The objective of this study was to determine true prevalence (TP) of bovine brucellosis [...] Read more.

In Ecuador, a national program for bovine brucellosis control has been in implementation since 2008. Given the costs, small- and medium-sized livestock holders are not completely committed to it. The objective of this study was to determine true prevalence (TP) of bovine brucellosis in small- and medium-sized herd populations, as well as the diagnostic sensitivity and specificity of the Rose Bengal (RB) test and the sero-agglutination test (SAT)-EDTA using a Bayesian approach. Between 2011 and 2016, 2733 cattle herds were visited, and 22,592 animal blood samples were taken in nineteen provinces on mainland Ecuador. Bayes-p and deviance information criterion (DIC) statistics were used to select models. Additionally, risk-factor analysis was used for herds according to their brucellosis test status. True prevalence (TP) in herds was estimated by pool testing. National seroprevalence of farms was 7.9% (95% CI: 6.79–9.03), and TP was 12.2% (95% CI: 7.8–17.9). Apparent prevalence (AP) in animals was 2.2% (95% CI: 1.82–2.67), and TP was 1.6% (95% CrI: 1.0–2.4). Similarly, the sensitivity of the RB was estimated at 64.6% (95% CrI: 42.6–85.3) and specificity at 98.9% (95% CrI: 98.6–99.0); for the SAT-EDTA test, sensitivity was 62.3% (95% CrI: 40.0–84.8) and 98.9% (95% CrI: 98.6–99.1) for specificity. Results of the two tests were highly correlated in infected and uninfected animals. Likewise, high spatial variation was observed, with the Coastal Region being the zone with the highest TP at 2.5%. (95% CrI: 1.3–3.8%) in individual animals and 28.2% (95% CI: 15.7–39.8) in herds. Risk factors include herd size, type of production (milk, beef, and mixed), abortions recorded, and vaccination. The results of this study serve to guide authorities to make decisions based on parallel testing at the beginning of a bovine brucellosis program for small livestock holders to increase sensitivity level of the screening tests in Ecuador. Full article

(This article belongs to the Special Issue Updates on Brucellosis)

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15 pages, 1707 KiB

Open AccessArticle

Seroprevalence and Risk Factors of Brucella Infection in Dairy Animals in Urban and Rural Areas of Bihar and Assam, India

by Ram Pratim Deka, Rajeswari Shome, Ian Dohoo, Ulf Magnusson, Delia Grace Randolph and Johanna F. Lindahl

Microorganisms 2021, 9(4), 783; https://doi.org/10.3390/microorganisms9040783 - 09 Apr 2021

Cited by 9 | Viewed by 2650

Abstract

This study assessed seropositivity of Brucella infection in dairy animals and risk factors associated with it. The cross-sectional study used multi-stage, random sampling in the states of Bihar and Assam in India. In total, 740 dairy animals belonging to 534 households of 52 [...] Read more.

This study assessed seropositivity of Brucella infection in dairy animals and risk factors associated with it. The cross-sectional study used multi-stage, random sampling in the states of Bihar and Assam in India. In total, 740 dairy animals belonging to 534 households of 52 villages were covered under this study. Serological testing was conducted by indirect enzyme-linked immunosorbent assay (iELISA). Animal-level Brucella seropositivity was found to be 15.9% in Assam and 0.3% in Bihar. Seropositivity in urban areas (18.7%) of Assam was found to be higher than in rural areas (12.4%). Bihar was excluded from the risk factor analysis, as only one Brucella seropositive sample was detected in the state. A total of 30 variables were studied for assessing risk factors, of which 15 were selected for multivariable regression analyses following a systematic process. Finally, only three risk factors were identified as statistically significant. It was found that animals belonging to districts having smaller-sized herds were less likely (p < 0.001) to be Brucella seropositive than animals belonging to districts having larger-sized herds. Furthermore, the chance of being Brucella seropositive increased (p = 0.007) with the increase in age of dairy animals, but decreased (p = 0.072) with the adoption of artificial insemination (AI) for breeding. We speculated that the identified risk factors in Assam likely explained the reason behind lower Brucella seropositivity in Bihar, and therefore any future brucellosis control program should focus on addressing these risk factors. Full article

(This article belongs to the Special Issue Updates on Brucellosis)

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14 pages, 2108 KiB

Open AccessArticle

Whole Genome Sequence Analysis of Brucella abortus Isolates from Various Regions of South Africa

by Maphuti Betty Ledwaba, Barbara Akorfa Glover, Itumeleng Matle, Giuseppe Profiti, Pier Luigi Martelli, Rita Casadio, Katiuscia Zilli, Anna Janowicz, Francesca Marotta, Giuliano Garofolo and Henriette van Heerden

Microorganisms 2021, 9(3), 570; https://doi.org/10.3390/microorganisms9030570 - 11 Mar 2021

Cited by 6 | Viewed by 2297

Abstract

The availability of whole genome sequences in public databases permits genome-wide comparative studies of various bacterial species. Whole genome sequence-single nucleotide polymorphisms (WGS-SNP) analysis has been used in recent studies and allows the discrimination of various Brucella species and strains. In the present [...] Read more.

The availability of whole genome sequences in public databases permits genome-wide comparative studies of various bacterial species. Whole genome sequence-single nucleotide polymorphisms (WGS-SNP) analysis has been used in recent studies and allows the discrimination of various Brucella species and strains. In the present study, 13 Brucella spp. strains from cattle of various locations in provinces of South Africa were typed and discriminated. WGS-SNP analysis indicated a maximum pairwise distance ranging from 4 to 77 single nucleotide polymorphisms (SNPs) between the South African Brucella abortus virulent field strains. Moreover, it was shown that the South African B. abortus strains grouped closely to B. abortus strains from Mozambique and Zimbabwe, as well as other Eurasian countries, such as Portugal and India. WGS-SNP analysis of South African B. abortus strains demonstrated that the same genotype circulated in one farm (Farm 1), whereas another farm (Farm 2) in the same province had two different genotypes. This indicated that brucellosis in South Africa spreads within the herd on some farms, whereas the introduction of infected animals is the mode of transmission on other farms. Three B. abortus vaccine S19 strains isolated from tissue and aborted material were identical, even though they originated from different herds and regions of South Africa. This might be due to the incorrect vaccination of animals older than the recommended age of 4–8 months or might be a problem associated with vaccine production. Full article

(This article belongs to the Special Issue Updates on Brucellosis)

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15 pages, 638 KiB

Open AccessArticle

Seroprevalence and Associated Risk Factors of Bovine Brucellosis at the Wildlife-Livestock-Human Interface in Rwanda

by Jean Bosco Ntivuguruzwa, Francis Babaman Kolo, Richard Simba Gashururu, Lydia Umurerwa, Charles Byaruhanga and Henriette van Heerden

Microorganisms 2020, 8(10), 1553; https://doi.org/10.3390/microorganisms8101553 - 09 Oct 2020

Cited by 11 | Viewed by 2998

Abstract

Bovine brucellosis is endemic in Rwanda; however, little information is available on seroprevalence and risk factors. Therefore, a cross-sectional study was conducted among cattle farmed at the wildlife-livestock-human interface (n = 1691) in five districts and one peri-urban district (n = [...] Read more.

Bovine brucellosis is endemic in Rwanda; however, little information is available on seroprevalence and risk factors. Therefore, a cross-sectional study was conducted among cattle farmed at the wildlife-livestock-human interface (n = 1691) in five districts and one peri-urban district (n = 216). Cattle were screened using the Rose Bengal test, then the results were confirmed by indirect enzyme-linked immunesorbent assay. Potential risk factors were determined with a questionnaire and analyzed for their association with seropositivity. In all districts, the animal and herd-level seroprevalence was 7.4% (141/1907) and 28.9% (61/212), respectively, 8.3% (141/1691) and 30.9% (61/198) at the interface, and 0.0% (0/216) in peri-urban areas. Among the potential risk factors, old age (≥5 years), cattle farmed close to wildlife, herds of cattle and small ruminants, history of abortions, and replacement animals were significantly associated with brucellosis (p < 0.05). Low awareness of zoonotic brucellosis, assisting calving without biosafety protection, drinking raw milk, and manual milking were each observed in more than 21.7% of cattle keepers whose herds were seropositive. This study confirmed brucellosis endemicity in cattle farmed close to wildlife in Rwanda, suggesting the need to focus control efforts in these areas. Educated farmers with a high awareness of zoonotic brucellosis had low bovine brucellosis seropositivity, which emphasizes the importance of education. Full article

(This article belongs to the Special Issue Updates on Brucellosis)

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12 pages, 1077 KiB

Open AccessArticle

Investigation of Brucella melitensis in Sable Antelope (Hippotragus niger) in South Africa

by Barbara Glover, Malcolm Macfarlane, Roy Bengis, Jacques O'Dell, Johan Steyl, Henriette van Heerden and Darrell Abernethy

Microorganisms 2020, 8(10), 1494; https://doi.org/10.3390/microorganisms8101494 - 29 Sep 2020

Cited by 7 | Viewed by 2597

Abstract

In this study, Brucella melitensis infection in sable antelope (Hippotragus niger) was investigated on two wildlife ranches in South Africa over a 12-year period in order to determine the origin of the outbreaks and the role of livestock in maintaining the [...] Read more.

In this study, Brucella melitensis infection in sable antelope (Hippotragus niger) was investigated on two wildlife ranches in South Africa over a 12-year period in order to determine the origin of the outbreaks and the role of livestock in maintaining the disease. Retrospective data were obtained from farm records and interviews as well as samples tested from different disease scenarios and clinical settings. On one ranch, 10 of 74 sable tested seropositive for B. melitensis in 2004 but were certified clear of infection after no further brucellosis cases were detected following repeated serological tests and culling over a five-year period. Recrudescence occurred in 2013 (7 of 187 brucellosis positives) and in 2014 (one positive), with persistent, latent infection being the most reasonable explanation. In a second case study, linked to the first one through a common vendor, 15 of 80 sable tested positive in 2016, some five years after the acquisition of the animals from a putative source. Brucella melitensis biovar 1 and/or 3 were isolated from each outbreak on both ranches. Both outbreaks resulted in substantial losses for the owners, arising from testing and culling and significant resource expenditure by the state. The study identified the diagnostic challenges for identifying and resolving disease outbreaks in wildlife, the persistence of B. melitensis in sable, the risks associated with animal movements, and the need for a wildlife-sensitive disease control scheme. Although the actual source of infection could not be identified, the investigation points away from local livestock as a source of ongoing infection while the persistent infection is consistent with the disease circulating within small, ranched populations and being spread through the keeping and trading of high-value animals. The implications of the study findings to disease control in wildlife are discussed. Full article

(This article belongs to the Special Issue Updates on Brucellosis)

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11 pages, 1829 KiB

Open AccessArticle

A Proof of Principle for the Detection of Viable Brucella spp. in Raw Milk by qPCR Targeting Bacteriophages

by Michaela Projahn, Jens A. Hammerl, Ralf Dieckmann and Sascha Al Dahouk

Microorganisms 2020, 8(9), 1326; https://doi.org/10.3390/microorganisms8091326 - 31 Aug 2020

Cited by 3 | Viewed by 2221

Abstract

Brucellosis is still a global health issue, and surveillance and control of this zoonotic disease in livestock remains a challenge. Human outbreaks are mainly linked to the consumption of unpasteurized dairy products. The detection of human pathogenic Brucella species in food of animal [...] Read more.

Brucellosis is still a global health issue, and surveillance and control of this zoonotic disease in livestock remains a challenge. Human outbreaks are mainly linked to the consumption of unpasteurized dairy products. The detection of human pathogenic Brucella species in food of animal origin is time-consuming and laborious. Bacteriophages are broadly applied to the typing of Brucella isolates from pure culture. Since phages intracellularly replicate to very high numbers, they can also be used as specific indicator organisms of their host bacteria. We developed a novel real-time PCR (qPCR) assay targeting the highly conserved helicase sequence harbored in all currently known Brucella-specific lytic phages. Quality and performance tests determined a limit of detection of <1 genomic copy/µL. In raw milk artificially contaminated with Brucella microti, Iz_v phages were reliably detected after 39 h of incubation, indicating the presence of viable bacteria. The qPCR assay showed high stability in the milk matrix and significantly shortened the time to diagnosis when compared to traditional culture-based techniques. Hence, our molecular assay is a reliable and sensitive method to analyze phage titers, may help to reduce the hands-on time needed for the screening of potentially contaminated food, and reveals infection risks without bacterial isolation. Full article

(This article belongs to the Special Issue Updates on Brucellosis)

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14 pages, 1963 KiB

Open AccessArticle

Seroprevalence and Molecular Identification of Brucella spp. in Camels in Egypt

by Aman Ullah Khan, Ashraf E. Sayour, Falk Melzer, Sherif Abdel Ghafar Elsayed El-Soally, Mandy C. Elschner, Waleed S. Shell, Amira A. Moawad, Shereen Aziz Mohamed, Ashraf Hendam, Uwe Roesler, Heinrich Neubauer and Hosny El-Adawy

Microorganisms 2020, 8(7), 1035; https://doi.org/10.3390/microorganisms8071035 - 13 Jul 2020

Cited by 20 | Viewed by 3836

Abstract

Brucellosis is one of the most important worldwide zoonoses of many countries including Egypt. Camel brucellosis has not gained much attention in Egypt yet. This study is focused on the three governorates with the highest camel populations and the largest camel markets in [...] Read more.

Brucellosis is one of the most important worldwide zoonoses of many countries including Egypt. Camel brucellosis has not gained much attention in Egypt yet. This study is focused on the three governorates with the highest camel populations and the largest camel markets in the country to determine the disease seroprevalence and identify the Brucella species in local camel holdings. In total, 381 serum samples were collected from male and female camels from Giza, Aswan, and Al-Bahr Al-Ahmar (the Red Sea) governorates. Samples were serologically examined using the Rose–Bengal plate test (RBPT), indirect ELISA (i-ELISA), competitive ELISA (c-ELISA) and complement fixation test (CFT). Brucella antibodies were detected in 59 (15.5%), 87 (22.8%), 77 (20.2%) and 118 (31.0%) of sera by RBPT, i-ELISA, c-ELISA and CFT, respectively. Using real-time PCR, Brucella DNA was amplified in 32 (8.4%) seropositive samples including Brucella abortus (25/32), Brucella suis (5/32) and Brucella melitensis (2/32), defining a complex epidemiological status. To the best of our knowledge, this is the first study reporting Brucella suis DNA in camel serum. The risk-associated factors including age, sex, breed and geographical distribution were statistically analyzed, showing non-significant association with seroprevalence. The results of this study will raise awareness for camel brucellosis and help develop effective control strategies. Full article

(This article belongs to the Special Issue Updates on Brucellosis)

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15 pages, 779 KiB

Open AccessPerspective

Proteomics of Brucella: Technologies and Their Applications for Basic Research and Medical Microbiology

by Gamal Wareth, Mathias W. Pletz, Heinrich Neubauer and Jayaseelan Murugaiyan

Microorganisms 2020, 8(5), 766; https://doi.org/10.3390/microorganisms8050766 - 20 May 2020

Cited by 13 | Viewed by 3683

Abstract

Brucellosis is a global zoonosis caused by Gram-negative, facultative intracellular bacteria of the genus Brucella (B.). Proteomics has been used to investigate a few B. melitensis and B. abortus strains, but data for other species and biovars are limited. Hence, a [...] Read more.

Brucellosis is a global zoonosis caused by Gram-negative, facultative intracellular bacteria of the genus Brucella (B.). Proteomics has been used to investigate a few B. melitensis and B. abortus strains, but data for other species and biovars are limited. Hence, a comprehensive analysis of proteomes will significantly contribute to understanding the enigmatic biology of brucellae. For direct identification and typing of Brucella, matrix-assisted laser desorption ionization—time of flight mass spectrometry (MALDI—TOF MS) has become a reliable tool for routine diagnosis due to its ease of handling, price and sensitivity highlighting the potential of proteome-based techniques. Proteome analysis will also help to overcome the historic but still notorious Brucella obstacles of infection medicine, the lack of safe and protective vaccines and sensitive serologic diagnostic tools by identifying the most efficient protein antigens. This perspective summarizes past and recent developments in Brucella proteomics with a focus on species identification and serodiagnosis. Future applications of proteomics in these fields are discussed. Full article

(This article belongs to the Special Issue Updates on Brucellosis)

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