Investigation on Brucella infection in farm animals in Saham, Sultanate of Oman with reference to human brucellosis outbreak

Abstract Background The objective of this study was to investigate Brucella infection in farm animals in Saham, Oman, with reference to a survey carried out by the Ministry of Agriculture & Fisheries (MAF) for Brucellosis during the period of May to July 2016 in Saham, following an outbreak of human brucellosis. We wanted to apply different serological, bacteriological and molecular tests in a time frame (phase 1, 2 & 3) with reference to the pivotal time of a human brucellosis outbreak to ascertain the status of the disease in Saham area where the MAF survey was conducted. Blood samples were collected from farm animals and sera were screened in parallel for Brucella antibodies using different serological tests. Results Using the RBT test, phase 1 sera showed seropositivity in sheep at 2.6%, (95% CI: 0.5–13.5%), in camel (5.9%, 1.1–27.0%), but not in sera from goats and cattle (0%). Using I-ELISA, seropositivity in goat was 3.1% (0.6–15.8%), with no positive sheep and cattle. Using c-ELISA for camel we found a seropositivity of 5.9% (1.1–27.0%). Furthermore, CFT seropositivity in goats was 21.9% (CI: 11.3–38.9), cattle and sheep sera were negative and camel was 5.9% (1.1–27.0%). In phase 2, the seropositivity in goats was 1.9% (1.4–2.6%), sheep 4.5% (3.5–5.8%), cattle 1.1%, (0.5–2.3%) and camels 18.2% (5.1–47.7%), Phase 3 sera were collected 6 months after the human brucellosis outbreak. With RBT, the seropositivity in goats was 3% (1.0–8.5%), sheep 2% (0.6–7.1%) cattle 1% (0.2–5.5%). With I-ELISA, goats & camels were negative, sheep were 3% (1.0–8.5%) and cattle 1% (0.2–5.5%). Moreover, B. melitensis was isolated from a bronchial lymph node of the RBT and I-ELISA seropositive cow and confirmed by Multiplex PCR and biochemical tests. Conclusion Using a retrospective study analysis of animal sera and following up after a human brucellosis outbreak, the present study showed a slight decrease in seropositivity of infected animals after the MAF implemented test and slaughter policy. The most interesting finding in this study was the isolation, identification and molecular characterization of Brucella melitensis in a cow (spillover), which is not a preferential host for Brucella melitensis.

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Incidental Laboratory Identification of Brucella melitensis in a case with Pancytopenia

International Journal of Current Microbiology and Applied Sciences ◽

10.20546/ijcmas.2021.1010.022 ◽

2021 ◽

Vol 10 (10) ◽

pp. 193-198

Author(s):

Mahalakshmi Kumaresan Lakshmi Shanmugam ◽

Ketan Priyadarshi Mahathi Gopalakrishnan ◽

Tamilarasu Kadhiravan Apurba Sankar Sastry

Keyword(s):

Bone Marrow ◽

Brucella Melitensis ◽

Public Health Problem ◽

Febrile Illness ◽

Human Brucellosis ◽

Automated Identification ◽

Serological Tests ◽

Test Culture ◽

Significant Public Health Problem ◽

Significant Public Health

Brucellosis is a bacterial zoonosis usually associated with exposure to infected animals or their products. Although a significant public health problem in India, exact prevalence and distribution are unknown owing to the imprecision of diagnosis and inadequacy of reporting and surveillance. Although the febrile illness is common, its manifestations are highly variable. Bone marrow suppression and consequent pancytopenia have been rarely reported. We present a case of 50 years old female diagnosed with human brucellosis associated with pancytopenia and non-specific clinical presentation, that was diagnosed incidentally on blood and bone-marrow culture. This was confirmed by serological tests like the standard agglutination test. Culture isolation using automated blood culture (e.g. BacT/ALERT), followed by identification using automated identification systems (e.g. MALDI-TOF and VITEK-2) help to reach accurate and timely diagnosis aiding in the management of the patient.

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Teknik Diagnostik Konvensional dan Lanjutan Untuk Pemeriksaan Mikrobiologi pada Infeksi Nosokomial di Indonesia

Jurnal Insan Cendekia ◽

10.35874/jic.v8i2.935 ◽

2021 ◽

Vol 8 (2) ◽

pp. 136-145

Author(s):

Maharani Pertiwi K. ◽

◽

Ayu Slatim Maifanda ◽

Amalia Ayu Febrianti ◽

Nabila Ina Zahra ◽

...

Keyword(s):

Nosocomial Infections ◽

Diagnostic Techniques ◽

Medical Laboratory ◽

Infectious Agents ◽

Biochemical Tests ◽

Serological Tests ◽

Molecular Tests ◽

Microbiological Examination ◽

Antigen Tests ◽

Pcr Techniques

Introduction: Nosocomial infections are infections caused by microbial such as bacteria, viruses, and fungi.that are acquired during the process of receiving health care. Diagnostic techniques for the examination of nosocomial infections play an important role in determining the accuracy of the infection of microorganisms causing infectious agents, so that the treatment given can be appropriate and minimize drug resistance. Purpose: This literature review is structured to provide an overview of diagnostic techniques fornosokomialinfection using conventional and advanced methods. Methods: The preparation of this review is based on the development of diagnostic techniques in the medical laboratory. Results: Conventional diagnostic techniques are generally carried out bymeans of culture on artificial media, macroscopic observations and biochemical tests. Further tests that can be applied are serological tests, antigen tests, and molecular tests such as PCR techniques. Conclusion: Conventional diagnostic techniques for microbiological examination of nosococomial infections require further tests to help establish a rapid and accurate diagnosis.

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Molecular detection of Brucella in milk using polymerase chain reaction

Czech Journal of Food Sciences ◽

10.17221/8318-cjfs ◽

2000 ◽

Vol 18 (No. 3) ◽

pp. 95-97 ◽

Cited By ~ 3

Author(s):

A. Mohsen

Keyword(s):

Direct Detection ◽

Brucella Melitensis ◽

Health Hazard ◽

Farm Animals ◽

Detection Methods ◽

Serological Tests ◽

Bacteriological Method ◽

Probe Assay ◽

Antigen Antibody ◽

Antibody Interactions

Brucellosis is a highly contagious disease affecting a wide variety of farm animals. It is also an important zoonosis, and man is often infected following contact with infected animals or the consumption of contaminated milk and milk products. At present, mainly bacteriological and serological detection methods are used. A bacteriological method takes days to weeks to grow the organism besides its health hazard. Serological tests are faster but antigen–antibody interactions can be faulted by non-specific interactions. A method for direct detection of Brucella melitensis in 1 ml of milk was developed on the basis of enzymatic treatment of milk components and subsequent PCR and line probe assay (LPA). After PCR, 3 × 104 CFU/ml sensitivity was obtained by agarose gel electrophoresis and LPA. The safety and sensitivity of LPA combined with its speed suggests the potential of this technique for diagnosis of brucellosis in milk rather than the time consuming classical methods.

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Serological and molecular investigation of human brucellosis in participants of Famenin brucellosis cohort study, Hamadan, Iran

Iranian Journal of Microbiology ◽

10.18502/ijm.v13i3.6394 ◽

2021 ◽

Author(s):

Maryam Adabi ◽

Manoochehr Karami ◽

Fariba Keramat ◽

Mohammad Yousef Alikhani ◽

Somaye Bakhtiari

Keyword(s):

Target Gene ◽

Brucella Melitensis ◽

Serological Test ◽

Human Brucellosis ◽

Intracellular Pathogen ◽

Pcr Assay ◽

Specific Primers ◽

Serological Tests ◽

Molecular Investigation ◽

Investigation Technique

Background and Objectives: Brucella is an intracellular pathogen that causes brucellosis in humans and animals. This study aimed to assess the results of brucellosis seroprevalence among participants of the Famenin brucellosis cohort with molecular investigation technique and determine Brucella-approved species. Materials and Methods: Following the first phase of the Famenin brucellosis cohort in 2016 which investigated the seroprevalence of brucellosis among 2367 participants in Famenin city, a total of 575 people including all seropositive and some seronegative people were examined again by wright serological tests in 2019. The PCR assay was accomplished on all cases that have wright titers ≥ 1/20 for tracing Brucella DNA using BCSP31 target gene and IS711 locus. Results: Out of 575 studied cases, 145 people had wright titers ≥ 1/20. The PCR reactions of these 145 blood samples were positive in 63/145 (43.44%) tested samples using primers (B4/B5) for Brucella genus detection. In the second PCR assay using specific-primers for Brucella abortus and Brucella melitensis, 18/63 (28.57%) of the samples were diagnosed as B. abortus, and 18/63 (28.57%) were diagnosed as B. melitensis. Conclusion: In this study, using the selected specific genes for the diagnosis of Brucella in the genus and species levels, the PCR technique was evaluated as a promising method for the rapid and safe detection of brucellosis besides the serological test for more accurate detection of brucellosis especially in cases that are not definitive.

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Fungal Diagnostics

Tutorial Topics in Infection for the Combined Infection Training Programme ◽

10.1093/oso/9780198801740.003.0017 ◽

2019 ◽

Author(s):

Shila Seaton ◽

Rohini J. Manuel

Keyword(s):

Fungal Pathogen ◽

Fungal Pathogens ◽

Histopathological Examination ◽

Clinical Specimen ◽

Fungal Disease ◽

Definitive Diagnosis ◽

Biochemical Tests ◽

Serological Tests ◽

Molecular Tests ◽

Circulating Antigens

The field of fungal diagnostics encompasses tests that are performed to help diagnose fungal disease, guide its management, and or monitor the effectiveness of its treatment. For some superficial skin and yeast infections, a clinical examination of the patient combined with microscopic examination of the sample may be sufficient to determine that fungal disease is present, even if the specific fungal pathogen is not identified. For deep- seated and systemic infections, a combination of diagnostic tests may be required in order to obtain a definitive diagnosis. These include microscopy to detect fungal elements, culture, detection of circulating antigens and antibodies, and molecular tests. More recently, molecular and proteomic approaches have increasingly dominated the conventional identification of pathogenic yeasts and, to some extent, filamentous fungi, since traditional methods are time consuming. More importantly, conventional methodologies have failed to identify common organisms that display uncharacteristic profiles, or fungal pathogens that are rarely encountered. The ‘gold standard’ for the definitive diagnosis of fungal disease is histology or culture of the fungal pathogen from a clinical specimen. A specimen will routinely be inoculated onto several different types of media, and then incubated at specific conditions and temperatures for up to twenty-one days. Media plates will be examined periodically for growth, and staff will try to identify the fungus using both macroscopic and microscopic morphologies. The few biochemical tests available, e.g. the urease test, can be helpful in identification, most often for yeast species. Microscopy of fungal isolates, histopathological examination of tissue, and fungal specific stains play fundamental roles in the diagnosis of infection for the variety of fungi that cause disease. The most common stain for identifying fungal elements from a cultured isolate is lactophenol fuschin/aniline blue stain. Figure 10.1 depicts the fruiting body (conidiophore) of Aspergillus fumigatus species complex, the most prevalent fungal species responsible for invasive aspergillosis (IA) in severely immunocompromised individuals. Figure 10.2 illustrates the phenotype of a three-day old colony. Serological tests are beneficial when non-culture based diagnosis of fungal disease is required. Complement fixation is predominantly used to diagnose endemic mycoses, e.g. coccidioidomycosis, blastomycosis, and histoplasmosis.

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Molecular characteristics of Brucella melitensis isolates from humans in Qinghai Province, China

Infectious Diseases of Poverty ◽

10.1186/s40249-021-00829-0 ◽

2021 ◽

Vol 10 (1) ◽

Author(s):

Zhi-Jun Zhao ◽

Ji-Quan Li ◽

Li Ma ◽

Hong-Mei Xue ◽

Xu-Xin Yang ◽

...

Keyword(s):

Tandem Repeats ◽

Brucella Melitensis ◽

Draft Genome ◽

Geographic Origin ◽

Variable Number ◽

Human Brucellosis ◽

Molecular Characteristics ◽

Whole Genome ◽

Qinghai Province ◽

Variable Number Tandem Repeats

Abstract Background The prevalence of human brucellosis in Qinghai Province of China has been increasing rapidly, with confirmed cases distributed across 31 counties. However, the epidemiology of brucellosis transmission has not been fully elucidated. To characterize the infecting strains isolated from humans, multiple-locus variable-number tandem repeats analysis (MLVA) and whole-genome single-nucleotide polymorphism (SNP)-based approaches were employed. Methods Strains were isolated from two males blood cultures that were confirmed Brucella melitensis positive following biotyping and MLVA. Genomic DNA was extracted from these two strains, and whole-genome sequencing was performed. Next, SNP-based phylogenetic analysis was performed to compare the two strains to 94 B. melitensis strains (complete genome and draft genome) retrieved from online databases. Results The two Brucella isolates were identified as B. melitensis biovar 3 (QH2019001 and QH2019005) following conventional biotyping and were found to have differences in their variable number tandem repeats (VNTRs) using MLVA-16. Phylogenetic examination assigned the 96 strains to five genotype groups, with QH2019001 and QH2019005 assigned to the same group, but different subgroups. Moreover, the QH2019005 strain was assigned to a new subgenotype, IIj, within genotype II. These findings were then combined to determine the geographic origin of the two Brucella strains. Conclusions Utilizing a whole-genome SNP-based approach enabled differences between the two B. melitensis strains to be more clearly resolved, and facilitated the elucidation of their different evolutionary histories. This approach also revealed that QH2019005 is a member of a new subgenotype (IIj) with an ancient origin in the eastern Mediterranean Sea.

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Systematic Review into Diagnostics for Post-Kala-Azar Dermal Leishmaniasis (PKDL)

Journal of Tropical Medicine ◽

10.1155/2013/150746 ◽

2013 ◽

Vol 2013 ◽

pp. 1-8 ◽

Cited By ~ 16

Author(s):

Emily R. Adams ◽

Inge Versteeg ◽

Mariska M. G. Leeflang

Keyword(s):

Diagnostic Accuracy ◽

Clinical Symptoms ◽

Statistical Modelling ◽

Study Quality ◽

Serological Tests ◽

Molecular Tests ◽

Kala Azar ◽

Valid Estimate ◽

Low Sensitivity ◽

Control Designs

Identification of post-kala-azar dermal leishmaniasis (PKDL) is important due to the long and toxic treatment and the fact that PKDL patients may serve as a reservoir for visceral leishmaniasis (VL). We summarized the published literature about the accuracy of diagnostic tests for PKDL. We searched Medline for eligible studies investigating the diagnostic accuracy of any test for PKDL. Study quality was assessed using QUADAS-2. Data were extracted from 21 articles including 43 separate studies. Twenty-seven studies evaluated serological tests (rK39 dipstick, ELISA, DAT, and leishmanin tests), six studies molecular tests, eight microscopy, and two cultures. Only a few of these studies reported a valid estimate of diagnostic accuracy, as most were case-control designs or used a reference standard with low sensitivity. The included studies were very heterogeneous, for example, due to a large variety of reference standards used. Hence, no summary estimates of sensitivity or specificity could be made. We recommend well-designed diagnostic accuracy trials that evaluate, side-by-side, all currently available diagnostics, including clinical symptoms, serological, antigen, molecular, and parasitological tests and possible use of statistical modelling to evaluate diagnostics when there is no suitable gold standard.

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Comparison of multiplex real-time polymerase chain reaction with serological tests and culture for diagnosing human brucellosis

Journal of Infection and Public Health ◽

10.1016/j.jiph.2018.11.008 ◽

2019 ◽

Vol 12 (3) ◽

pp. 337-342 ◽

Cited By ~ 7

Author(s):

Tuba Dal ◽

Soner Sertan Kara ◽

Aytekin Cikman ◽

Cigdem Eda Balkan ◽

Ziya Cibali Acıkgoz ◽

...

Keyword(s):

Polymerase Chain Reaction ◽

Real Time ◽

Human Brucellosis ◽

Chain Reaction ◽

Serological Tests ◽

Polymerase Chain

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The first outbreak of brucellosis in the region of Sabac

Vojnosanitetski pregled ◽

10.2298/vsp1008634m ◽

2010 ◽

Vol 67 (8) ◽

pp. 634-637 ◽

Cited By ~ 1

Author(s):

Ljiljana Markovic-Denic ◽

Vesna Skodric-Trifunovic ◽

Vladimir Zugic ◽

Dragana Radojcic ◽

Goran Stevanovic

Keyword(s):

Confidence Interval ◽

Animal Species ◽

Epidemiological Methods ◽

Human Brucellosis ◽

Reference Laboratory ◽

Serological Tests ◽

Military Medical Academy ◽

Medical Academy ◽

Laboratory Confirmation ◽

The Military

Background/Aim. In Serbia brucellosis is a primary disease of the animals in the southern parts of the country. The aim of this study was to describe the first outbreak of human and animal brucellosis in the region of Sabac, Serbia. Methods. An epidemiological investigation was conducted to identify a source of outbreak and the ways of transmission of brucellosis infection in human population. A descriptive and analytical epidemiological methods (cohort study) were used. Additional data included monthly reports of the infectious diseases from the Institutes of Public Health and data from the Veterinary Specialistic Institute in Sabac. The serological tests for human brucellosis cases were performed in the Laboratory of the Military Medical Academy; laboratory confirmation of animal brucellosis cases was obtained from the reference laboratory of the Faculty of Veterinary Medicine, Belgrade. Results. Twelve cases of brucellosis were recorded from February 9 to September 1, 2004. Total attack rate was 8.1% (7.5% of males, 14.2% of females). Relative risk (RR) of milk consumption was 8.9 (95% confidence interval: 1.63-13.38), and RR for direct contact with animals was 14 (95% confidence interval: 3.5-55.6). The prevalence of seropositive animals in 33 villages of the Macva region accounted for 0.8%. Regarding animal species, sheep were predominant - 264 (95.7%). Out of a total number of seropositive animals, ELISA results were positive in 228 (88.7%) of them. Conclusion. As contact epidemics generally last longer, it is probable that the implemented measures of outbreak control did reduce the length of their duration.

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Seroprevalance of Toxoplasmosis in sheep and goat: Iraq/ Sulaimania

The Iraqi Journal of Veterinary Medicine ◽

10.30539/iraqijvm.v35i1.599 ◽

2011 ◽

Vol 35 (1) ◽

pp. 16-24

Author(s):

Lazem H. Al-Taie

Keyword(s):

Age Groups ◽

Neural System ◽

Farm Animals ◽

Economic Losses ◽

Age Group ◽

Serum Samples ◽

Serological Tests ◽

Back Ground ◽

Qualitative Determination

Back ground: Toxoplasmosis is an important zoonosis that causes economic losses in animal herds due to abortion and stillbirth as well as changes in the reproductive and neural system of susceptible animals . Objective: The aims of the present study is to determination the prevalence of T. gondii in farm animals ( sheep& goat)of both genders and different ages in Sulaimani province by using two serological tests (ELISA and LAT). Methods: Blood samples were collected from farm animals ,142 sheep and 46 goats , of different sexes and ages. Tow different serological tests ,ELISA and LAT for qualitative determination of T. gondii antibody titer in sheep and goats serum samples. Results: The prevalence rate in sheep was 73 (51.7 %) and 82 (57 %) , and 21 (54.6 %) and 25 (54.35 %) in goats ,by ELISA and LAT respectively. The prevalence of toxoplasmosis was highest in age group 7-9 (66.6%) in sheep in compares’ with other age groups. There was no significant differences between both spp.and tow test. Conclusion: Statistical results show no significant differences between both tests (ELISA &LAT) at (P ≥ 0.05).The prevalence of toxoplasmosis was increased proportionally with the age of animals, while gender has no effect on the prevalent rate .

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