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An overview of the epidemiology and epizootology of brucellosis in selected countries of Central and Southeast Europe.

The objective of this paper is to give an overview of the epidemiologic and epizootic status of brucellosis in selected countries of Central and Southeast Europe (Balkan region). Based on dimension of the disease problem, there is a need to establish collaboration in the eradication and prevention of brucellosis between all countries in the region. Although there were no readily accessible data concerning epidemiology and epizootology of brucellosis in these countries, the limited official and published data were analyzed. The incidence of brucellosis caused by Brucella melitensis in sheep, goats and humans is a very significant problem in Macedonia and Greece. In Greece, cattle are also affected either by B. melitensis or B. abortus. The disease is an endemic problem in some regions of Yugoslavia and includes B. suis biovar 2 in pigs and in Croatia, B. melitensis in sheep, goats and human is found occasionally. No problem appears to exist with brucellosis in Bulgaria. Financial well-supported brucellosis control programs of the European Union that will include all countries, regardless of the magnitude of brucellosis incidence, are needed for eradication and control of brucellosis.

Animals↗

Field investigations of brucellosis in cattle and small ruminants in Syria, 1990-1996.

The authors present the epidemiological status of brucellosis in cattle and small ruminants in Syria from 1990 to 1996, based on laboratory findings at the Brucellosis Centre, Damascus. Initial investigations using the Rose Bengal plate test, the complement fixation test and a miniaturised variant of the slow agglutination test were conducted throughout the country in 1990 and 1991, revealing an overall herd seroprevalence rate of 3.14% in cattle herds and 2.94% in small ruminant flocks. Although partially biased by previous vaccination of young female cattle with S19 vaccine, these figures indicate that brucellosis in cattle is widespread, particularly in the urban governorates (provinces) of Damascus, Aleppo and Suwaydah. Brucellosis seroprevalence in sheep and goats was relatively high in the governorates of Damascus, Aleppo and Dara'a. The results of a second series of investigations, performed between 1992 and 1996, show that herd seroprevalence in cattle decreased steadily from 17.48% in 1992, to 2.59% in 1996, in the Government-owned farms, while seroprevalence increased in the private sector during the same period. The difference may be explained by the restriction of brucellosis vaccination to public farms (although this was far from systematic), combined with partial application of a 'test-and-slaughter' policy. In sheep and goats, brucellosis seroprevalence fluctuated in the two sectors, but was higher in the private sector where husbandry is principally extensive. Bacteriological investigations led to the isolation of Brucella melitensis biovars 2 and 3 in sheep and B. abortus biovar 9 in cattle. Although no specific methodology was employed, particularly with regard to sampling, this study is significant as the first international report of the distribution of brucellosis in Syria. Further, well-structured studies are required, the results of which could be used to plan an appropriate national control programme for brucellosis.

Abortion, Veterinary↗

TLR4 polymorphism in Iranian patients with brucellosis.

OBJECTIVE: Brucellosis is a zoonosis of both public health and economic significance in most developing countries. Polymorphisms in Toll-like receptor-4 (TLR4) have been reported to be associated with a blunted immune response to microbial pathogens. Information regarding any association between genetic variation of TLR4 and susceptibility to brucellosis is not available in the literatures. The main purpose of this research is to evaluate the role of polymorphic alleles of TLR4 gene in susceptibility to brucellosis. MATERIALS AND METHODS: In this case-control study, 198 patients with brucellosis and 111 healthy volunteers matched for sex, age and geographic area were evaluated by genotyping for polymorphism in TLR4 gene (Asp299Gly) using amplification refractory mutation system (ARMS)-PCR method. RESULTS: Allele 896G was more prevalent in patients with brucellosis compared to healthy controls (33.6% vs. 20.7%, P=0.000003). Also the frequency of G allele of TLR4 gene was significantly higher in male patients with brucellosis compared to the same sex in control group (36% vs. 21.7%, P=0.00005). Multiple logistic regression analysis demonstrated that male patients heterozygous at allele G gene had a significantly higher risk for brucellosis with an odds ratio of OR 2.89, 95% CI: 1.79-4.69, P<0.0001). CONCLUSION: This study is the first to show an association between genetic polymorphism in TLR4 gene and susceptibility to brucellosis.

Adolescent↗

Specific IgE response in patients with brucellosis.

In the search to find discriminative serological markers to differentiate between patients with acute brucellosis and those with chronic brucellosis, an enzyme-linked immunosorbent assay (ELISA) was used to determine and compare the brucella-specific IgE response in 80 sera from patients with acute brucellosis, 37 sera from patients with chronic brucellosis, 26 sera from patients with positive blood cultures for bacteria other than brucella and 51 sera from healthy controls. The IgE findings were compared to brucella-specific IgG, IgM, IgA and IgG1-4 demonstrated by ELISA, and to microagglutination test (MAT) results. Elevated (positive) antibrucella IgE titres were detected in 89 and 81% of sera from patients with acute and chronic brucellosis respectively. The predominant antibodies found in patients with acute brucellosis were of the IgG, IgM, IgA, IgE, IgG1 and IgG3 types while in chronic brucellosis IgG, IgA, IgE and IgG4 were found. Although IgE can be detected in patients with brucellosis, it does not discriminate between the acute and chronic stages of the disease.

Acute Disease↗

From the discovery of the Malta fever's agent to the discovery of a marine mammal reservoir, brucellosis has continuously been a re-emerging zoonosis.

Brucellosis is not a sustainable disease in humans. The source of human infection always resides in domestic or wild animal reservoirs. The routes of infection are multiple: food-borne, occupational or recreational, linked to travel and even to bioterrorism. New Brucella strains or species may emerge and existing Brucella species adapt to changing social, cultural, travel and agricultural environment. Brucella melitensis is the most important zoonotic agent, followed by Brucella abortus and Brucella suis. This correlates with the fact that worldwide, the control of bovine brucellosis (due to B. abortus) has been achieved to a greater extent than the control of sheep and goat brucellosis (due to B. melitensis), these latter species being the most important domestic animals in many developing countries. The long duration and high cost of treatment of human brucellosis reduces the efficacy of the therapy. There is no human vaccine for brucellosis and the occurrence of brucellosis is directly linked to the status of animal brucellosis in a region. In this context, the Word Health Organization has defined the development of a human vaccine, besides the implementation of control and eradication programs in animals, as a high priority. The pathogenicity for humans of B. suis biovars 1, 3 and 4 is well established, whereas B. suis biovar 2 seems to be less pathogenic. Indeed, although hunters and pig farmers have repeatably experienced infectious contact with B. suis biovar 2 (found in wild boar and outdoor-rearing pigs in Europe), isolation of B. suis biovar 2 from human samples have only been seldom reported. Marine mammal brucellosis, due to two new proposed Brucella species i.e. B. cetaceae and B. pinnipediae, represents a new zoonotic threat but the pathogenicity for humans of the different Brucella species found in cetaceans and pinnipeds still has to be clearly established.

Animals↗

Increased serum levels of interferon-gamma and interleukin-12 during human brucellosis.

Brucellosis is a zoonotic disease that often becomes chronic with a high rate of recurrence. To understand the cytokines induced during this infection we determined the levels of interleukin (IL)-1beta, IL-2, IL-4, IL-8, IL-12, tumor necrosis factor-of (TNF-alpha) and interferon-gamma (IFN-gamma) in serum of patients with brucellosis and compared with those without brucellosis and controls. Control sera were taken from healthy persons residing in an area that was not endemic for brucellosis. The levels of IL-1, IL-2, IL-4, and TNF-alpha were not detectable in all the sera from patients. There was no difference in the level of IL-8 in patients with brucellosis and those without this disease. However, IL-8 was significantly higher in these two groups of patients compared with the controls. Significantly higher levels of IL-12 and IFN-alpha were found in the serum of patients with brucellosis compared with patients without brucellosis and controls. These data indicate that there is induction of Thl type cytokines during human brucellosis.

Adolescent↗

Brucellosis as a world problem.

Brucellosis is one of the most widespread and economically the most ravaging of zoonoses. The occurrence of the acute, often incapacitating infection in man caused by Brucella melitensis usually coincides with occurrence of the infection in sheep and goats. Well-known foci of this infection have been identified in the Mediterranean basin, Central Asia and Latin America. Recent investigations in parts of Africa and India have shown that the infection is much more widespread than was previously suspected and that more foci remain to be discovered by the application of laboratory methods in epidemiological investigations. Bovine infection has a wider distribution and is more important than infection of sheep and goats as far as economic losses are concerned. Although the infection has been reduced by control measures to a low level of incidence in some countries of Europe and North America, its incidence in other parts of the world has actually increased because of emphasis on increased animal production and aggregation under poor hygienic conditions. This is particularly the case with dairy production units which have developed around rapidly growing urban centres in many developing countries. Although human infection with B. abortus may be mild, it can cause troublesome and intractable illness. The economic ravages of bovine brucellosis are tremendous and the losses in Latin America and the USA alone have been estimated by their respective governments to be 700 million dollars annually. The world figure must be truly staggering. Swine brucellosis is largely a problem of the Americas although a few foci have recently been discovered in other parts of the world, notably in Europe and Southern Asia. In the Americas, swine infection is second only to bovine brucellosis in its economic importance. A large number of human cases of B. suis infection are detected every year in the endemic areas. Other forms of brucellosis, e.g. B. canis in dogs and B. suis, biotype 4 in reindeer, have a limited distribution and are responsible for only sporadic human infections. B. ovis of sheep is more widespread but is generally confined to sheep. Annexes 1 and 2 show the reported incidence of brucellosis in man and animals. However, the officially reported data are generally incomplete and the actual incidence must in most cases be much higher than is shown in these tables. FAO and WHO have been assisting countries in which brucellosis is endemic by organizing control measures, providing training and reference materials, and by focusing coordinated research on problems which arise in these programmes. Some of the highlights of this work are outlined in the following paragraphs.

Animals↗

The epidemiological situation of brucellosis in Africa.

Brucellosis, as a geomedical literature survey has shown, is spread to a higher of lesser degree of incidence virtually over the whole African continent. Its epidemiological spectrum of host animals is especially rich amongst the domestic and wild animal species. All domestic animal species and 21 of 26 herbivorous and 5 of 12 carnivorous wild animal species screened yielded positive brucella antibody serotitres. Thus self-maintaining natural foci independent of the domestic stock have firmly been established. The mapping of the epidemiological situation must be based on survey data which should have been planned, exactly performed, analyzed and reported according to statistical rules. Unfortunately Africa is especially badly covered in the world literature in the field of brucellosis. Representing itself 32.0% (49/153) of the countries of the world, their Medical and Veterinary Departments contributed only 2.1% of the computerized world literature on brucellosis in the period 1969-74. This picture can be considerably changed by including the non-computerized "grey literature" of Department and Laboratory Annual Reports into the screening process. By introducing objective, mathematical criteria for determining the degree of incidence from the available data, the following results were obtained from the literature survey: brucellosis is known or suspected to exist in 40 of the 49 African countries (82%). In 20 (41%) of these countries it represents a major problem, in 10 (20.4%) a moderate problem in the other 10 countries a minor problem to human health and economy. Following the rule that brucellosis can be expected to exist everywhere in the world (with the exception of a few islands) where cattle, sheep and/or goats are kept unless it has been eliminated by vigourous measures of control, it may be expected that the remaining 9 African countries not yet reporting also harbour this chronic, zoonotic disease. 35 of the 40 countries reporting have found brucellosis regularly, 1 exceptionally among domestic animals, 5 among game animals. Only 17 of these 35 (48.5%) have scheduled the disease, even if its incidence has been reported to be high. Only 32 of the 49 countries report on human brucellosis. The main reservoirs of disease and risks of infection vary from place to place with the different main kinds of domestic stock, the animal husbandry and human consumption methods and habits. Thus mainly the cattle herds are the source of infection for those countries where large numbers of cattle are kept and the drinking of raw milk is the custom; however, there is no strict correlation between these two factors. Goats and sheep fill the epidemiological place in all those countries where the lack of marketable milk curtails its consumption; the small ruminants are kept as the main source of meat supply and in close contact with the farmers family within the compound or house.

Africa↗

Human brucellosis in a nonendemic country: a report from Germany, 2002 and 2003.

Human brucellosis has become a rare disease in Germany since the eradication of bovine and ovine/caprine brucellosis in this country. Therefore, most physicians are unfamiliar with the illnesses clinical presentation, diagnostic tools, and therapeutic strategies. This retrospective study was carried out to evaluate the epidemiological, clinical, and laboratory features of human brucellosis in Germany in the years 2002 and 2003. Thirty-one bacterial isolates from 30 patients sent to the German national reference laboratory were characterized using the genus-specific bcsp31 real-time PCR, the species-specific AMOS-PCR, and standard microbiological methods for the detection and identification of Brucella spp. The medical records of all patients with bacteriologically confirmed brucellosis were evaluated. All 31 isolates proved to be Brucella (30 Brucella melitensis and 1 Brucella suis). Most of the brucellosis patients were infected in endemic countries while visiting friends and relatives during their summer holidays. One case of laboratory-acquired infection was identified. Brucellosis was transmitted mainly by the consumption of contaminated unpasteurized milk or cheese from goats and sheep. The patients presented primarily with flu-like symptoms, i.e. fever, chills, sweating, headaches, arthralgia, and myalgia. In most cases, however, symptoms and signs of focal complications, e.g. spondylitis, endocarditis, and meningoencephalitis, predominated. The rate of complications was much higher than that in endemic countries, presumably as a result of diagnostic delay due to a low index of suspicion. In summary, physicians in nonendemic countries such as Germany must be aware of brucellosis being a possible cause of fever of unknown origin in immigrants and tourists travelling from endemic countries.

Adolescent↗

Pulmonary involvement in brucellosis.

OBJECTIVES: Pulmonary involvement is a rare manifestation of brucellosis. The aim of this study was to determine the incidence and forms of pulmonary involvement in the course of brucellosis. METHODS: A prospective study was carried out in 110 patients with brucellosis. All the patients were evaluated with their pulmonary symptoms, physical examination and chest radiography. If pulmonary pathologic findings were present, patients underwent additional diagnostic evaluations including computerized tomography of the thorax and pulmonary function tests. RESULTS: From 110 patients, 11 (six females and five males) were diagnosed as pulmonary brucellosis. Eight of 11 patients had pulmonary symptoms including cough, sputum and dyspnoea. Radiologic findings were parenchymal nodules, lobar pneumonia, paratracheal lymphadenopathy and pleural effusion. At the end of the treatment of brucellosis, clinical findings of pulmonary involvement were recovered in all patients except four dyspnoeic patients who had coexisting COPD. Radiological findings were normal in three and improved in four patients after 6 months of the treatment. CONCLUSIONS: Pulmonary involvement is a rare event in the course of brucellosis. But especially in endemic regions, brucellosis should never be forgotten as a causative agent in patients with pulmonary symptoms.

Adult↗

Diagnosis of human brucellosis with ELISA.

ELISA tests for total (IgG + IgM + IgA), IgG, and IgM anti-Brucella antibodies, which utilised only commercially available reagents, were used to diagnose human brucellosis. Assays for total antibodies in sera from 22 patients with confirmed acute brucellosis, 1 patient with probable acute brucellosis, and 3 patients with probable chronic brucellosis gave readings that were more than double those found in hundreds of control sera. All sera from patients with acute and chronic brucellosis had significantly elevated IgG levels. Although there were a few acute patients with IgM levels only slightly higher than those of some controls, most patients with acute disease could readily be differentiated from both the non-brucellosis patients and patients with chronic brucellosis by measuring macroglobulins. Both the IgG and IgM levels of sera from acute patients persisted for at least 8 months. The results of this show that ELISA is an excellent method for screening large populations for Brucella antibodies and for differentiation between the acute and chronic phases of the disease.

Acute Disease↗

Importance of screening household members of acute brucellosis cases in endemic areas.

Isolated reports of brucellosis among family members have been documented. The aim of this study is to determine if active serological screening of the households' members of acute brucellosis cases will detect additional unrecognized cases. From May 2000 to October 2001, patients with acute brucellosis were enrolled and their household members were serologically screened for brucellosis using the Standard Agglutination Test (SAT). Fifty-five index cases with acute brucellosis and 404 household members were enrolled. The majority of index cases (48%) were young adults, and 79% were illiterate. Ownership of animals and ingestion of unpasteurized raw milk were reported by 45 and 75% of the index cases respectively. Of the 55 families screened, 23 (42%) had two family members or more with serological evidence of brucellosis and 32 (58%) had only the index case. Households of > or = 5 members and a history of raw-milk ingestion by family members were risk factors associated with the seropositives (P < 0.05). Of the 404 household members screened, 53 (13%) were seropositive; of these 39 (74%) were symptomatic, and 9 (35%) had brucella bacteraemia. Symptomatic seropositives tended to have bacteraemia and higher brucella antibody titres compared to asymptomatic seropositives (P < or = 0.05). Screening family members of an index case of acute brucellosis will detect additional cases.

Acute Disease↗

Rapid diagnosis of human brucellosis by peripheral-blood PCR assay.

A single-step PCR assay with genus-specific primers for the amplification of a 223-bp region of the sequence encoding a 31-kDa immunogenetic Brucella abortus protein (BCSP31) was used for the rapid diagnosis of human brucellosis. We examined peripheral blood from 47 patients, with a total of 50 cases of brucellosis, and a group of 60 control subjects, composed of patients with febrile syndromes of several etiologies other than brucellosis, asymptomatic subjects seropositive for Brucella antibodies, and healthy subjects. Diagnosis of brucellosis was established in 35 cases (70%) by isolation of Brucella in blood culture and in the other 15 cases (30%) by clinical and serological means. The sensitivity of our PCR assay was 100%, since it correctly identified all 50 cases of brucellosis, regardless of the duration of the disease, the positivity of the blood culture, or the presence of focal forms. The specificity of the test was 98.3%, and the only false-positive result was for a patient who had had brucellosis 2 months before and possibly had a self-limited relapse. In those patients who relapsed, the results of our PCR assay were positive for both the initial infection and the relapse, becoming negative once the relapse treatment was completed and remaining negative in the follow-up tests at 2, 4, and 6 months. In conclusion, these results suggest that the PCR assay is rapid and easy to perform and highly sensitive and specific, and it may therefore be considered a useful tool for diagnosis of human brucellosis.

Adolescent↗

Ocular complications associated with brucellosis in an endemic area.

PURPOSE: To report the ocular manifestations associated with brucellosis in an endemic area. METHODS: We prospectively evaluated 147 patients with the diagnosis of brucellosis between May 1996 to May 2000 and recorded the ocular and systemic findings. The diagnosis was based on clinical findings, positive serological and bacteriological tests (Brucella agglutination test: over 1/160 titer, blood culture). RESULTS: Thirty-eight patients (26.0%) with brucellosis had ocular complications: conjunctivitis in 26 (17.7%), anterior uveitis in six (4.1%), posterior uveitis in one (0.7%), dacryoadenitis in two (1.4%), episcleritis in three (2.1%). Three of the seven patients with uveitis had spondylitis associated with brucellosis. Osteoarticular complications in brucellosis were more frequent in the patients with ocular involvement though the difference was not statistically significant compared with patients without ocular involvement. CONCLUSIONS: Ocular manifestations are frequent in brucellosis so an ophthalmologic examination should be routinely performed in patients with brucellosis in endemic areas.

Adolescent↗

Effect of the treatment of brucellosis on leukocyte superoxide dismutase activity and plasma nitric oxide level.

BACKGROUND: The mechanisms by which brucellae evade intracellular killing by polymorphonuclear leukocytes are incompletely understood. In this study, we evaluated changes of leukocyte superoxide dismutase (SOD) activity and plasma total nitrite as an indicator of nitric oxide (NO) levels during brucellosis therapy. METHODS: Thirty-two patients with acute brucellosis, 27 patients with chronic brucellosis and 30 healthy controls were included in the study. Patients with acute brucellosis were tested for leukocyte SOD activity and plasma total nitrite levels before, during (21st day), and at the end (45th day) of the combined therapy of rifampicin and doxycycline. The same parameters were also investigated in chronic cases and controls. RESULTS: The SOD activities were lower in patients with acute brucellosis before therapy compared with those 21 and 45 days after starting therapy (P < 0.001). In contrast, total nitrite levels did not change significantly (P > 0.05). CONCLUSIONS: In the present study, leukocyte SOD activity was found to be decreased in patients with acute brucellosis. Enzyme activity was increased by treatment, finally reaching the activity of healthy controls. Using an antioxidant agent in addition to classical antimicrobial therapy for acute brucellosis might be a therapeutic approach.

Acute Disease↗

Human brucellosis: review of an under-diagnosed animal transmitted disease.

Human brucellosis is an important animal transmitted disease of man. Although, the cases have been recorded all over the world, the prevalence is higher in developing countries. Lack of sufficient knowledge about the disease among the physicians, its under-diagnosis or misdiagnosis and absence of effective prevention and management strategies are attributed to the widespread of the disease. Increase in the occurrence of animal brucellosis has also resulted indirectly in an increase in the prevalence of human infection. Absence of characteristic clinical symptoms, chronic nature of the infection and difficulty in isolation of the causal agent from the patients make the diagnosis of the disease more difficult. The serological tests employed for diagnosing human brucellosis vary in terms of their sensitivity and specificity. Therefore, a combination of serological tests is desirable. Currently no vaccine is available against human brucellosis, which could check the spread of the disease effectively. It is suggested that clinicians investigate the cases of pyrexia of unknown origin (PUO) for brucellosis. It is desirable that specimens from cases of tuberculosis, typhoid, rheumatoid arthritis, urogenital infections, kala-azar, cirrhosis, bacterial endocarditis, leukemia and filariasis should also be screened for brucellosis in man. The cases of meningitis of unestablished etiology as the cases of human brucellosis are often misdiagnosed as cases of typhoid or tuberculosis.

Animals↗

Human brucellosis in the Mid-West 2002-3.

Human brucellosis remains a serious public health issue in Ireland. Clinical notifications in the Mid-Western Area (HSE-MWA) underestimate the burden of illness and attendant morbidity in the region. The diagnosis of acute and chronic human brucellosis depends on the clinical evidence and the results from laboratory serological testing or culture on rare occasion. This study examined the clinical evidence behind locally defined serological "positives" in the HSE-MWA from 2002 to 2003. Ninety cases were detected in 2002 and 31 in 2003. While sampling bias is likely to be present, aspects of brucellosis in Ireland were confirmed. Middle-aged males were most commonly affected. The majority of cases were linked to farming or veterinary practice. Symptoms such as sweats, fever and weight loss were commonly associated with acute brucellosis infection while malaise was common in acute and chronic brucellosis. A clear definition of what is notifiable is needed. Surveillance systems must appreciate the importance of both clinical and laboratory evidence to classify confirmed or probable brucellosis as paired sera were not common. Public health authorities must follow-up the clinical aspects for accurate national statistics. General practitioners in the Mid-West appear to be vigilant regarding brucellosis in their patients. Regional zoonoses committees are useful in monitoring disease prevalence in human and animal populations without compromising confidentiality.

Adolescent↗

[Comparison of the serological tests used for the laboratory diagnosis of brucellosis].

Brucellosis which is an important public health problem is a zoonotic disease that causes economic loss, and seen all over the world as well as in our country. The aim of this study was to compare the results of Rose Bengal test, standard tube agglutination test (STA), Coombs tube agglutination test, Rivanol tube agglutination test and ELISA (IgA, IgG and IgM) method, in patients who were suspected to have brucellosis. Blood and serum samples collected from 77 patients were included to the study. Blood cultures were performed from all of the patients and Brucella spp were isolated in 35 (45.4%) of them. At least one of the serological tests for Brucella was found positive in 62 samples (80.5%). When culture positivity accepted as the gold standard, the sensitivities of the tests were calculated as follows; 100% for Rose Bengal, 94.3% for STA, 97.1% for ELISA IgG, 94.3 for ELISA IgA and 71.4 for ELISA IgM. No prezone phenomenon was observed in the samples. Brucella Coombs tube agglutination test did not increase the chance of diagnosis of brucellosis, unlike the results of previous studies. We found that Rose Bengal and STA tests were still efficient methods for brucellosis serodiagnosis. The results of rivanol tube agglutination test were similar to STA test, and this test can be used especially in patients with brucellosis relapse. ELISA test detecting IgA and IgG antibody titers together found to be effective methods increasing the chance of brucellosis diagnosis, and could be helpful for follow up of brucellosis.

Agglutination Tests↗