Unpasteurized Dairy Products Research Articles

Disseminated, fatal reactivation of bovine tuberculosis in a patient treated with adalimumab: a case report and review of the literature.

Tumor necrosis factor inhibitors (TNFi) are known to increase the risk of tuberculosis (TB) reactivation, though cases involving Mycobacterium bovis are rarely reported. We describe a case of disseminated TB with M. bovis in a 78-year-old woman with a negative Interferon-Gamma-Release Assay (IGRA), taking adalimumab due to rheumatoid polyarthritis, which resulted in a fatal outcome. The atypical clinical and histopathological features were initially interpreted as sarcoidosis. The case occurred in Switzerland, an officially bovine tuberculosis-free country. The whole genome sequence of the patient's cultured M. bovis isolate was identified as belonging to the animal lineage La1.2, the main genotype in continental Europe, but showed significant genetic distance from previously sequenced Swiss cattle strains. In a literature review, four cases of bovine tuberculosis reactivation under TNFi treatment were identified, with pulmonal, oral and intestinal manifestations. Similar to our patient, two cases presented a negative IGRA before TNFi initiation, which later converted to positive upon symptomatic presentation of M. bovis infection. This case highlights the diagnostic challenges of TB in immunosuppressed patients, the limited sensitivity of IGRA, and the importance of considering TB reactivation even in regions declared free of bovine tuberculosis. Detailed patient histories, including potential exposure to unpasteurized dairy products, are essential for guiding preventive TB treatment before TNFi initiation.

Delayed diagnosis of persistent Q fever: a case series from China

BackgroundQ fever, caused by the zoonotic pathogen Coxiella burnetii, exhibits a worldwide prevalence. In China, Q fever is not recognized as a notifiable disease, and the disease is overlooked and underestimated in clinical practice, leading to diagnostic challenges.Case presentationWe present a case series of three patients diagnosed with persistent Q fever between 2022 and 2023. The average age of our three cases was 63.33 years old, consisting of two males and one female. The medical history of the individuals included previous valve replacement, aneurysm followed by aortic stent-graft placement and prosthetic hip joint replacement. At the onset of the disease, only one case exhibited acute fever, while the remaining two cases were devoid of any acute symptoms. The etiology was initially overlooked until metagenomic next-generation sequencing test identified Coxiella burnetii from the blood or biopsy samples. Delayed diagnosis was noted, with a duration ranging from three months to one year between the onset of the disease and its confirmation. The epidemiological history uncovered that none of the three cases had direct exposure to domestic animals or consumption of unpasteurized dairy products. Case 1 and 2 resided in urban areas, while Case 3 was a rural resident engaged in farming. All patients received combination therapy of doxycycline and hydroxychloroquine, and no recurrence of the disease was observed during the follow-up period.ConclusionQ fever is rarely diagnosed and reported in clinical practice in our country. We should be aware of persistent Q fever in high-risk population, even with unremarkable exposure history. Metagenomic next-generation sequencing holds great potential as a diagnostic tool for identifying rare and fastidious pathogens such as Coxiella burnetii.

Epidemiological study of brucellosis (Malta fever) in the city of Torbat Jam from 2017 to 2022

Introduction: Brucellosis, a globally prevalent disease, affects both humans and animals, with notable significance in Iran. While the severity and mortality of this disease are rare, it poses a substantial economic burden due to reduced productivity and livestock abortion. Torbat Jam city has a high prevalence of brucellosis in the region, necessitating an investigation into its epidemiological aspects. Objectives: The present study aimed to investigate the epidemiology of brucellosis in Torbat Jam city. Materials and Methods: This study examined 5366 cases of brucellosis in the city of Torbat Jam from 2017 to 2022. A comprehensive checklist encompassing variables, such as age, gender, place of residence, occupation, season, history of contact with infected animals, consumption of unpasteurized dairy products, and vaccination history was used for data collection. The t-test was used to compare the means of two groups, while the chi-square test was employed to assess the relationship between categorical variables and grouped numerical variables. A significance level of 0.05 was considered in this study. Results: A total of 5366 cases of Malta fever were analyzed in this study, with male patients comprising 50.40% and female patients accounting for 49.60% of the cases. The mean age of the patients during the study period was 33.18±18.81 years. Brucellosis onset occurred at an average age of 35.39±18.03 years in males and 30.96±19.01 years in females, with a statistically significant difference (P value <0.01). The youngest reported case of brucellosis was 1 year old, while the oldest was 101 years old. The highest disease occurrence (60.8%) was observed in the age group of 15-50 years, with a higher prevalence observed in both genders within this age group. Conclusion: The city of Torbat Jam is considered a high-risk area in terms of brucellosis prevalence. Factors such as rural living, contact with animals and consumption of unpasteurized dairy products significantly contribute to the disease’s spread in these regions. The results underscore the insufficient public awareness regarding brucellosis transmission and prevention, highlighting the urgent need for educational programs to enhance knowledge and awareness among individuals in these areas.

Exposure of cattle to tick-borne encephalitis virus in the historical endemic zone in north-eastern France

BackgroundTick-borne encephalitis (TBE) is a severe human neuroinfection caused by TBE virus (TBEV). TBEV is transmitted by tick bites and by the consumption of unpasteurized dairy products from infected asymptomatic ruminants. In France, several food-borne transmission events have been reported since 2020, raising the question of the level of exposure of domestic ungulates to TBEV. In this study, our objectives were (i) to estimate TBEV seroprevalence and quantify antibodies titres in cattle in the historical endemic area of TBEV in France using the micro virus neutralisation test (MNT) and (ii) to compare the performance of two veterinary cELISA kits with MNT for detecting anti-TBEV antibodies in cattle in various epidemiological contexts. A total of 344 cattle sera from four grid cells of 100 km² in Alsace-Lorraine (endemic region) and 84 from western France, assumed to be TBEV-free, were investigated.ResultsIn Alsace-Lorraine, cattle were exposed to the virus with an overall estimated seroprevalence of 57.6% (95% CI: 52.1–62.8%, n = 344), varying locally from 29.9% (95% CI: 21.0–40.0%) to 92.1% (95% CI: 84.5–96.8%). Seroprevalence did not increase with age, with one- to three-year-old cattle being as highly exposed as older ones, suggesting a short-life duration of antibodies. The proportion of sera with MNT titres lower than 1:40 per grid cell decreased with increased seroprevalence. Both cELISA kits showed high specificity (> 90%) and low sensitivity (less than 78.1%) compared with MNT. Sensitivity was lower for sera with neutralising antibodies titres below 1:40, suggesting that sensitivity of these tests varied with local virus circulation intensity.ConclusionsOur results highlight that cattle were highly exposed to TBEV. Screening strategy and serological tests should be carefully chosen according to the purpose of the serological study and with regard to the limitations of each method.

Estimation of Total Proteins and Minerals Content of Bovine Brucellosis and Anaplasmosis in Some Local Dairy Farms in Kosti, Sudan

Bovine brucellosis, caused by Brucella abortus, is a significant bacterial infectious zoonotic disease not only poses a threat to animal health but also presents economic challenges and public health concerns due to its potential transmission to humans. Humans can contract the disease through direct contact with infected animals or consumption of unpasteurized dairy products. Bovine anaplasmosis is a haemolytic disease of cattle caused by the bacterium Anaplasma marginale which can cause adult mortality, abortion, weight loss, and a reduction in performance. This paper described the results obtained from the effects of brucellosis and anaplasmosis on serum proteins and the level of the macro elements (Calcium, Phosphorus, Potassium, Sodium, and Magnesium). These minerals play an essential role in animal metabolism. Out of fourteen Bovine serum samples, five samples were found to be positive for Brucella (35.72%) and four samples were found to be positive for Anaplasma (28.57%), the remaining samples were negative. Total protein (Tb) and (Alb) levels were significantly increased (P<0.01) in both brucellosis and anaplasmosis cattle compared with controls. On the other hand Globulin (Glob) was significantly decreased (P<0.01) in brucellosis cattle compared to the controls, also the present study showed that, concentration of Na, Ca, and Mg minerals were significantly increased in case of Brucellosis compared to the controls and significantly low (P<0.05) in potassium level, while animals infected with anaplasmosis showed an increased level of Na and K and decreased level in Mg. In summary, comprehension of laboratory medicine parameters is crucial for monitoring animal health and diagnosing diseases like brucellosis and anaplasmosis. These parameters are essential for implementing effective control measures and ensuring livestock well-being. The objective of this study was to estimate the total proteins and minerals content of Bovine Brucellosis and Anaplasmosis in ...

(245) Epididymo-Orchitis as the First Presentation of Brucellosis in an Endemic Area

Abstract Introduction Brucellosis, which is also called Mediterranean or Malta fever, is an endemic enzootic disease and can involve various organ systems. It is commonly endemic in the Mediterranean countries and the Middle East. Epididymo-orchitis is a focal form of human brucellosis. Brucella species cause granulomatous orchitis usually presenting as an acute or chronic unilateral swelling of the testis. Patients rarely present to the clinicians with acute scrotum due to Brucellosis Epididymo-Orchitis (BEO) as an initial finding. Objective Our aim is to estimate the prevalence of the BEO among the cases diagnosed with Epididymoorchitis in our hospital over the last four years. Methods Retrospective study from 2015–2018 involving all cases diagnosed with Epididymoorchitis in our hospital. A total of 92 cases of Epididymoorchitis were diagnosed during this period. These cases presented to ER complaining of severe unilateral scrotal pain, swelling, fever and sweating. Local examination showed swollen enlarged testis and epididymis with marked tenderness. Brucella serologic test for both B abortus and B melitensis were requested routinely for all cases of epididymo-orchitis. Scrotal Doppler ultrasonographic examination was requested at initial diagnosis and after 2 weeks to exclude abscess formation. Results Out of 92 cases, 8 were diagnosed to have brucellosis (8.7%). The median age of patients was 32 years (range, 18–41 years). All patients had positive history of consuming unpasteurized dairy products, which is a risk factor for brucellosis. The diagnosis of brucellosis was made via positive serological testing results (Brucella abortus and melitensis). One case showed testicular abscess formation. All cases were given combination of tetracyclin and rifampicin for 6–12 weeks. Two cases required prolonged hospital stay due to testicular abscess formation in one and development of Brucellosis spondylitis in the other. Gentamycin was added for one week in such cases. Conclusions In endemic regions for brucellosis, we have to consider it in the workup of all cases of epididiymo-orchitis. The diagnosis is easy by positive serology tests. Positive cases need special antibiotic combination protocol for longer time and close monitoring for development of local complications or other systemic brucellosis manifestations Disclosure No.

Ayurvedic Insights into Re-Emerging Human Brucellosis

Brucellosis is a bacterial infection caused by Brucella melintensis. It is one of the most widespread endemic zoonotic diseases present globally, more prevalent in developing countries. It is transmitted to humans either through direct contact with infected animal tissues or fluids or indirectly by ingesting contaminated meat and unpasteurized dairy products. The route of transmission includes mainly mucocutaneous. Farmers, butchers, hunters, and veterinarians are mainly at higher risk of contracting this condition. Mostly the affected individuals exhibit flu-like symptoms, remittent fever, malaise, arthralgia, and weight loss which lasts from a few days to several years. The condition is often misdiagnosed due to nonspecific clinical symptoms, leading to inadequate treatment and prolonged illness leading to severe complications like endocarditis. Currently, there is no vaccine for human brucellosis. The modern management includes administration of antibiotics. The gold standard diagnostic tool is direct bacteriological testing. Ayurvedic understanding of endemic diseases highlights the unique concept of Janapathhodhwamsa proposed by Acharyas. They considered Vayu, Jala, Kala, and Desa as common factors in manifestation of disease. Any alteration in these factors can be understood as imbalance and disease outbreak emerged due to urbanization, lack of hygienic measures in animal husbandry, etc. remaining a public health hazard. Brucellosis is known by its undulant nature which closely resembles the Vishama Jwara in classics which is a Sannipatika type of Jwara. Amapachana, Shodhana, and Samana line of management followed by Rasayana, Satvritta Palana as a preventive line can be adopted to manage Brucellosis.

Isolation, Identification and Detection of Campylobacter Fetus from Dairy & Poultry Products and Development of PCR

Campylobacters spp. are mainly responsible for gastrointestinal infections. The main source of infection is the consumption of contaminated or undercooked meat (especially poultry products), unpasteurized dairy products and untreated water. It is a type of zoonotic disease where it spreads through feces and mouth. Campylobacter species are widespread among warm-blooded animals, so they are found in food animals such as poultry, cattle, pigs, sheep and ostriches. Therefore, animal samples were used in this study. The most common causes of this infection are Campylobacter jejuni (C.jejuni), Campylobacter coli (C.coli) and Campylobacter fetus (C. fetus). Molecular techniques such as multiplex polymerase chain reaction (mPCR) have been optimized to detect these species. Campylobacter fetus was initially divided into two subspecies to reconcile significant epidemiological, clinical, and biochemical differences between the two similar taxa, which were later shown to be highly related at the genomic level. Campylobacter fetus occurs mainly in the intestines of cattle and sheep and can cause spontaneous abortion in both animals.

Exploring in vivo and in vitro infection models in brucellosis research: A mini-review

Brucellosis is a serious disease that affects both animals and humans. It is caused by consuming unpasteurized dairy products that are contaminated with the Brucella bacteria. To study the pathobiology of this disease and develop preventive strategies, researchers rely on in vivo and in vitro models. A systematic literature search was conducted in January 2024, which revealed 38 studies that used these models in the previous four years. Mice were the most commonly used model for studying the disease's virulence genes, immune responses, vaccination, and treatment testing. Out of the 38 articles discussing infection models in brucellae, 6 used only in vivo models, 9 used only in vitro models, and 24 used both models. In addition, there were 32 studies with in vitro experiments, most of which utilized macrophages to study intracellular survival mechanisms and host-pathogen interactions. The studies mainly focused on B. abortus, as it had a significant impact on public and livestock health. Both in vivo and in vitro models were used to understand comprehensive intracellular mechanisms, immune responses, and treatment evaluations. However, there were several challenges in using these models, such as ethical concerns and host pathogen-specific immune responses. While both models provided important insights, the final selection choice of the model mostly depended on the research objectives, pathogen type, and availability of resources. Nevertheless, validation and understanding of these models are important to predict responses in the natural hosts

Characterization of the adaptive cellular and humoral immune responses to persistent colonization of Brucella abortus strain RB51 in a Jersey cow.

Brucella abortus strain RB51 is the commercial cattle vaccine used in the United States (US) and many parts of the world against bovine brucellosis. RB51 was licensed for use in 1996, and it has been shown to be safe and efficacious in cattle, eliciting humoral and cellular responses in calves and adult animals. In 2017, an epidemiological trace-back investigation performed by the Centers for Disease Control and Prevention (CDC) identified human cases of brucellosis caused by infection with RB51. These infections resulted from the consumption of unpasteurized dairy products, which were traced back to otherwise healthy animals that were shedding RB51 in their milk. At the current time, six adult Jersey cows have been identified in the U.S. that are shedding RB51 in milk. One of the RB51 shedding cattle was obtained and housed at the National Animal Disease Center (NADC) for further study. Improved understanding of host cellular and humoral immune responses to RB51 in persistently colonized cattle may be achieved by the characterization of responses in shedding animals. We hypothesized, based on the lack of RB51 clearance, that the RB51 shedder animal has a diminished adaptive cellular immune response to RB51. Our data demonstrate that in the presence of persistent RB51 infection, there is a lack of peripheral anti-RB51 CD4+ T cell responses and a concurrently high anti-RB51 IgG humoral response. By understanding the mechanisms that result in RB51 persistence, the development of improved interventions or vaccinations for brucellosis may be facilitated, which would provide public health benefits, including reducing the risks associated with the consumption of non-pasteurized milk products.

Anadoluda Süt Ürünleri ve Pediatrik Bruselloz

Introduction: The purpose of the study is to evaluate the follow-up and treatment outcomes of pediatric brucellosis cases who presented to our participating centers during the term and were diagnosed with brucellosis. Methods: The study, conducted with the support of participating centers in the Central Anatolia region, was completed in about 2 years. The results of patients diagnosed with brucellosis between 2018 and 2021 were retrospectively scanned. During this screening, patients with missing results in the system were excluded from the study. Results: According to the data collected from the participating centers, we included a total of 68 patients in our study who met our criteria after excluding those with incomplete information. These patients were between the ages of 5 and 17, with a mean age of 8.7±3.1 years. Our analysis of the patients' demographics revealed that they mostly resided in rural areas where unpasteurized milk and dairy products were commonly consumed. Although there were some patients from higher socioeconomic backgrounds who purchased organic products from local markets, the vast majority (90%) had a history of consuming unpasteurized milk and its by-products. In addition, a significant proportion (50%) of patients from rural areas had a family history of brucellosis. The most frequently reported symptom among patients was joint pain, which was present in 83% of cases. Elevated liver enzymes were the most common laboratory finding, affecting 32% of patients, while thrombocytopenia was the least common. The preferred treatment regimen for patients was doxycycline and rifampicin. Although aminoglycoside treatment was initially favored for inpatients (80%), it was not commonly prescribed for outpatients. Our analysis of the data revealed that only two patients experienced a relapse of the disease. While the overall treatment outcomes were successful, there was no statistically significant association between treatment preferences and disease recurrence. Conclusion: Every year, thousands of children in our country visit pediatric clinics complaining about joint pain. These patients are usually examined for extensive investigations. It should be remembered that with suspicious patient history and demography, brucellosis may be the answer under the symptoms, accompanied by elevated liver enzymes and cytopenia.

Sacroileitis Developing in Irregularly Treated Brusellosis

Brucellosis; Fever, also known as corrugated fever, is a zoonotic disease charactarized by chills and muscle aches. It can be seen endemic in many countries including our country. Consumption of unpasteurized dairy products is the most common mode of transmission. Diagnosis is made by culture of the organism and increase in antibody titer in serum samples. Complications affecting many systems, mostly osteoarticular, are seen in the disease. In our case, a case of sacroileitis developed as a result of irregular treatment in a patient who was engaged in animal husbandry and was diagnosed with brucellosis is presented. In our case, it was started that osteoarticular involvement could develop under treatment and the importance of regular treatment at correct doses was emphasized.

Evaluation of Prevalence of Hepatitis E Clinical Markers among Donors in Estonia.

Hepatitis E virus (HEV) is now considered the most common cause of acute hepatitis worldwide. There are no published data about the prevalence of antibodies to HEV and RNA in donor sera in Estonia, and this precludes planning measures for preventing HEV proliferation through blood transfusion services. Here, were report data from an analysis of 1002 sera on the prevalence of anti-HEV IgG and IgM and the viral RNA. The antibodies were found in 48 donor sera (4.8%); of these, 40 (4%) harbored anti-HEV IgG, 15 (1.5%) contained anti-HEV IgM, and 7 donors had anti-HEV antibodies of both classes simultaneously. HEV RNA was not detected in any blood serum. Statistical associations of infection risk factors (gender, age, travel in the last six months, contact with pigs and/or wild boars in the last six months, consumption of thermally unprocessed/raw pork or boar meat, raw/unfiltered tap water or water from natural sources, unpasteurized farm dairy products, and unwashed berries and/or vegetables) were assessed. None of the listed factors were found to be associated with a higher or lower risk of anti-HEV antibody presence. At the same time, an increasing share of anti-HEV IgG carriers with age was found. The absence of HEV RNA in the analyzed donor plasma samples proves that HEV acute infection prevalence in Estonia does not exceed the average level of European countries. There is no urgent necessity to enter a requirement for a total screening of blood plasma for HEV RNA prevalence in Estonia.

SAT609 Panhypopituitarism Secondary To Pituitary Abscess In A Patient With Neurobrucellosis

Abstract Disclosure: G. De la Peña Sosa: None. A. Cabello Hernández: None. M. Gómez Sámano: None. F. Gómez Pérez: None. Background: Pituitary abscess is a rare but potentially life-threatening disease that could lead to panhypopituitarism. It is important to suspect the diagnosis, especially in regions of the world with endemic infectious diseases. Clinical Case: A 55-year, farmer, with a history of consumption of unpasteurized dairy products, began in February 2023 with headache, impaired consciousness and fever. Initial tests were consistent with neuroinfection: lumbar puncture with cerebrospinal fluid (CSF) with elevated proteins (>200 mg/dL, n <45 mg/dL), decreased glucose (14 mg/dL with a serum glucose of 76 mg/dl), pleocytosis (800 mm3, n <5 mm3). Head CT-scan did not report alterations. Antibiotic treatment was started due to diagnosis of acute meningitis. Later Brain MRI was performed showing ventriculitis; the pituitary gland was heterogeneous with the presence of an 8x8mm abscess.Pituitary axis hormones were evaluated, showing results compatible with panhypopituitarism with central hypothyroidism (normal serum TSH 1.05 mIU/L (n 0.3-5 mIU/L), decreased free T4 0.57 ng/dL (n 0.63-1.34 nd/dL), decreased total T3 0.47 ng/mL (n 0.64-1.81 ng/mL)), central hypocortisolism (decreased morning cortisol levels 5.72 mcg/dL (n 6.7-22.6 mcg/dL) and ACTH 20 pg/mL (n 10-50 pg/ml)), central hypogonadism (decreased LH <0.20 mUI/mL (n 1.24-862 mUI/mL), decreased FSH 0.37 mUI/mL (n 1.27-19.26 mUI/mL), decreased testosterone 0.24 ng/mL (n 1.75-7.81 ng/mL)), growth hormone deficiency (decreased IGF-1 9.99 ng/mL (n 61-206 ng/mL)). Hormone replacement treatment with hydrocortisone and levothyroxine was started. Due to the epidemiological background, Rose Bengal test for Brucella spp. was performed, with positive results (n negative) and 2-mercaptoethanol Brucella agglutination test was also positive >1:20 (n <1:20). Diagnosis of neurobrucellosis was made and treatment with doxycycline, rifampin and ceftriaxone was started. Neurosurgery service suggested conservative management with antibiotics. Patient showed improvement in his neurological status and infectious symptoms disappeared. Subsequent pituitary MRI showed pituitary intraglandular abscess with a 15% decrease.The patient was discharged six weeks later with prednisone, levothyroxine, somatropin and testosterone, as well as doxycycline and rifampin for another 4 months. Conclusion: Pituitary abscess should be considered in a differential diagnosis of patients with neuro infection symptoms, panhypopituitarism and heterogenous image in the MRI. Opportune medical management may result in a lower mortality and a higher recovery of pituitary hormone function. Reference: Karagiannis, Apostolos K A et al. “Pituitary abscess: a case report and review of the literature.” Endocrinology, diabetes & metabolism case reports vol. 2016 (2016): 160014. doi:10.1530/EDM-16-0014 Presentation: Saturday, June 17, 2023

Retrospective Evaluation of Patients with Brucellosis: Five Years of Experience

Objective: We aimed to evaluate the epidemiological, clinical features, laboratory findings, and treatment regimens of patients with brucellosis in the Tokat region. Methods: In this cross-sectional study, the data of 236 adult patients followed with brucellosis in Tokat Gaziosmanpaşa University Medical Faculty Hospital between January 2016 and December 2021 were evaluated retrospectively. Diagnostic criteria of brucellosis were determined as culture positivity, or positivity in the Wright test at ≥ 1/160 titration, or a 4-fold increase in titer of the Wright test repeated at 2 – 3 week intervals. Demographic data, symptoms, physical examination findings, and laboratory results of the patients were evaluated. Results: 72% (n=170) of the patients were male, and the mean age was 44.47±16.64. 53% (n=125) were engaged in animal husbandry, 21.6% (n=51) consumed unpasteurized dairy products, and 13.1% (n=31) had a family member with brucellosis. Seven (2.9%) patients with a prediagnosis of Crimean-Congo hemorrhagic fever (CCHF) were diagnosed with acute brucellosis. The most common symptoms were muscle-joint pain (70.3%), fever (50.8%), and sweating (43.2%). Spondylodiscitis was observed in 13.6%, epididymal-orchitis in 8.2%, and neurobrucellosis in 0.4% of the patients. Blood culture positivity was detected in 7.2% of the cases. 91.5% had elevated C-reactive protein (CRP) levels, and 48.3% had elevated erythrocyte sedimentation rates. The most common hematological findings were anemia (28%), leukopenia (10.6%), and thrombocytopenia (8.5%). As a therapy doxycycline + rifampicin combination was started at 36%, doxycycline + gentamicin to 5.8%, doxycycline + streptomycin to 5.4%, and doxycycline + rifampicin + gentamicin to 29.7% of the patients. Alternative treatment regimens were given to 22.9% (n=54) of the patients. Conclusion: Brucellosis is still a critical zoonotic disease in our region. It should be kept in mind when diagnosing patients with fever, muscle-joint pain, and sweating, with epidemiological histories, have elevated acute phase reactants and cytopenia. It should also be remembered that brucellosis can be confused with CCHF in endemic areas.

Rapid and Specific Detection of B. melitensis Targeting BMEI1661 Gene Using Loop-mediated Isothermal Amplification (LAMP) Combined With Lateral Flow immunoassay (LFIA).

B. melitensis is the most pathogenic zoonotic species of Brucella transmitted to animals through fetal secretions, placenta, and vaginal discharges of infected animals and humans by ingesting unpasteurized milk, dairy products, and raw meat. Early detection of B. melitensis is essential for timely intervention and control of the disease. The gold standard diagnostic methods, such as culture, are time-consuming and may take several weeks aiding to the disease spread. Loop-mediated isothermal amplification assay (LAMP) is widely used to detect infectious pathogens. LAMP can be utilized as a rapid point-of-care test, but has lower specificity which can be enhanced by combining this test with lateral flow immunoassay. No point-of-care test is available for detecting Brucella melitensis in clinical samples. Herein, we developed a LAMP coupled with lateral flow immunoassay (LFIA) for the specific detection of B. melitensis. The sensitivity of LAMP-LFIA was found to be 12.1fg of genomic DNA isolated from the organism, which is 100-fold more sensitive to conventional PCR and equally sensitive to Real-time (RT-PCR). Moreover, the assay demonstrated high specificity when tested against other Brucella and non-Brucella species. The infective dose of B. melitensis is relatively low for humans, which may remain undetected by conventional PCR, but will be detected using the new technique.

Detection of Toxoplasma gondii Infection in Small Ruminants: Old Problems, and Current Solutions.

Toxoplasmosis is a parasitic zoonosis of veterinary importance, with implications for public health. Toxoplasma gondii infection causes abortion or congenital disease in small ruminants. Moreover, the consumption of infected meat, cured meat products, or unpasteurized milk and dairy products can facilitate zoonotic transmission. Serological studies conducted in various European countries have shown the high seroprevalence of specific anti-T. gondii antibodies in sheep and goats related to the presence of oocysts in the environment, as well as climatic conditions. This article presents the current status of the detection possibilities for T. gondii infection in small ruminants and their milk. Serological testing is considered the most practical method for diagnosing toxoplasmosis; therefore, many studies have shown that recombinant antigens as single proteins, mixtures of various antigens, or chimeric proteins can be successfully used as an alternative to Toxoplasma lysate antigens (TLA). Several assays based on DNA amplification have been developed as alternative diagnostic methods, which are especially useful when serodiagnosis is not possible, e.g., the detection of intrauterine T. gondii infection when the fetus is not immunocompetent. These techniques employ multicopy sequences highly conserved among different strains of T. gondii in conventional, nested, competitive, and quantitative reverse transcriptase-PCR.

Seroprevalence of <i>Brucella</i> infection in cattle and small ruminants in South Omo zone, southern Ethiopia

A cross-sectional study was conducted in selected districts of the South Omo Zone to estimate the seroprevalence of brucellosis and its associated risk factors. Additionally, the knowledge, attitude, and practices (KAP) of livestock keepers about the disease were also assessed. A total of 1349 sera samples were collected from 450 cattle and 899 small ruminants (450 goats and 449 sheep) kept under an extensive production system. Rose Bengal Plate Test was used for screening and ELISA as a confirmatory test for the detection of antibodies against Brucella species. Based on confirmatory tests, the overall seroprevalence of brucellosis was 2.2 % (95% CI: 1.1 – 4.1%) in cattle, 2.0% (95% CI: 0.9, 3.8%) in goats and 1.3% (95% CI: 0.5, 2.9%) in sheep with higher seropositivity in cattle compared to small ruminants. Seropositivity did not vary significantly (p> 0.05) with agroecology, age, and sex groups in cattle. However, a higher seroprevalence of 2.7% was detected in male cattle compared to 1.6% for females. Seroprevalence was higher in small ruminants from the lowland agroecology (3.0%) than those from the midlands (0.8%). Study respondents had a low level of knowledge on brucellosis with only 43% of them having some information about the disease. Most of the respondents have routinely engaged in practices that can expose them to infections such as assisted delivery (65%), contact with after-birth materials (50%), and handling aborted materials (15%) without any protection. High consumption of unpasteurized dairy products (93%) such as raw whole milk and traditionally fermented dairy product is also common. This study provides information on the occurrence f brucellosis in major livestock species kept at cross-border marginal areas with limited disease information. The existence of various exposure practices implies the need for creating awareness among livestock keepers on general disease transmission and its zoonotic role.

The thick-borne encephalitis caused by alimentary transmission - case series report.

Tick-borne encephalitis (TBE) is a central nervous system zoonotic disease transmitted by ticks. Tick-borne encephalitis virus (TBEV) is one of the main causes of lymphocytic meningitis in the areas of its endemic occurrence. A mode of transmission of TBEV which is rarely observed in clinical practice is an alimentary transmission through consumption of unpasteurised dairy products from infected animals. The following article contains detailed description of the clinical course of TBE among five family members, for whom the occurrence of TBE was temporarily associated with the consumption of unpasteurised goat's milk from the same source. The epidemiological outbreak presented in this article is the fifth ever described case of the milk-borne TBE in Poland. More so, the clinical course of the disease has shown differences from the typical course characterised so far in the literature. Clinical cases of TBE described in this study were similar to infections caused by tick bites in humans. The following article discusses available methods of preventing TBE, with emphasis on alimentary transmission of TBEV, since possibility of serious detrimental long-term neurological complications resulting from TBE was stressed in prior literature.

Severe Streptococcus equi Subspecies zooepidemicus Outbreak from Unpasteurized Dairy Product Consumption, Italy

During November 2021-May 2022, we identified 37 clinical cases of Streptococcus equi subspecies zooepidemicus infections in central Italy. Epidemiologic investigations and whole-genome sequencing showed unpasteurized fresh dairy products were the outbreak source. Early diagnosis by using sequencing technology prevented the spread of life-threatening S. equi subsp. zooepidemicus infections.