Animal Brucellosis Research Articles

Some Risk Factors Associated with Animal Brucellosis in Aswan Governorate, Egypt

Some Risk Factors Associated with Animal Brucellosis in Aswan Governorate, Egypt

Fluorescence Polarization Assay for Infection Diagnostics: A Review

Rapid and specific diagnosis is necessary for both the treatment and prevention of infectious diseases. Bacteria and viruses that enter the bloodstream can trigger a strong immune response in infected animals and humans. The fluorescence polarization assay (FPA) is a rapid and accurate method for detecting specific antibodies in the blood that are produced in response to infection. One of the first examples of FPA is the non-competitive test for detecting brucellosis in animals, which was followed by the development of other protocols for detecting various infections. Fluorescently labeled polysaccharides (in the case of brucellosis and salmonellosis) or specific peptides (in the case of tuberculosis and salmonellosis, etc.) can be used as biorecognition elements for detecting infections. The availability of new laboratory equipment and mobile devices for fluorescence polarization measurements outside the laboratory has stimulated the development of new fluorescence polarization assays (FPAs) and the emergence of commercial kits on the market for the detection of brucellosis, tuberculosis, and equine infectious anemia viruses. It has been shown that, in addition to antibodies, the FPA method can detect both viruses and nucleic acids. The development of more specific and sensitive biomarkers is essential for the diagnosis of infections and therapy monitoring. This review summarizes studies published between 2003 and 2023 that focus on the detection of infections using FPA. Furthermore, it demonstrates the potential for using new biorecognition elements (e.g., aptamers, proteins, peptides) and the combined use of FPA with new technologies, such as PCR and CRISPR/Cas12a systems, for detecting various infectious agents.

Molecular Prevalence, Phylogenetic Analysis, and PCR-based Detection of Brucella melitensis in Humans and Cattle in Southern Khyber Pakhtunkhwa, Pakistan

Brucellosis is a bacterial disease due to Brucella melitensis, considered a zoonotic agent affecting humans and animals, especially in areas with high disease occurrence, south Khyber Pakhtunkhwa, Pakistan. This work was designed to evaluate the molecular rate of B. melitensis in humans and cattle species and also to perform a phylogenetic analysis between both species. A cross-sectional survey involving 800 participants, including 600 cattle and 200 human participants, underwent blood sample collection with conventional PCR and IS711 locus PCR amplification and Sanger sequencing. The findings detected Brucella melitensis in 37 of the 800 samples, with a molecular prevalence of 3.1% in cattle and 9% in humans. The molecular trees play a role in zoonotic transmission and point to the necessity of a further unified approach toward the management of brucellosis in both humans and animals. This is further backed by the use of 95% C.I for the prevalence rates making the results statistically robust. This research shows that using the IS711 insertion sequence is an efficient and selective method for identifying Brucella species.

Genomic analysis of Brucella isolates from animals and humans, Türkiye, 2010 to 2020.

BackgroundBrucellosis is a bacterial zoonosis causing severe illness in humans and animals and leading to economic losses in the livestock production in Türkiye and other endemic countries.AimWe aimed at investigating genomic differences of Brucella isolates from animals and humans in Türkiye.MethodsWe used whole genome sequencing (WGS) to assess the genetic diversity of Brucella isolates from 41 provinces in Türkiye and compared with isolates from other countries. We applied allele-based typing and core genome single nucleotide polymorphism (cgSNP) determination.ResultsOf the 106 Turkish Brucella isolates included, 57 were B. abortus and 49 were B. melitensis. One B. melitensis and two B. abortus isolates were identified as vaccine strains. Most (n = 55) B. abortus isolates clustered in three major branches, with no spatial discernible pattern. Of the B. melitensis isolates, 48 were assigned to the Eastern Mediterranean lineage with no discernible patterns between host species, location and sampling date. The Turkish isolates clustered with isolates from neighbouring countries such as Greece and Syria, but some also with isolates from human patients in European countries, like Germany, Norway and Sweden, suggesting that the source may be travel-related.ConclusionSeveral B. melitensis and B. abortus lineages are circulating in Türkiye. To decrease the prevalence and prevent brucellosis in animals and humans, stricter control measures are needed, particularly in areas where humans and animals have close contact. Furthermore, illegal transportation of animals across borders should be more closely controlled and regulated.

Epidemiological Investigation of Animal Brucellosis in Domestic Ruminants in Greece from 2015 to 2022 and Genetic Characterization of Prevalent Strains.

Brucellosis is one of the most important zoonotic diseases in Greece, causing a significant burden on both human and animal vitality as well as economic loss. The present study was conducted from 2015 to 2022 on 711,415 serum samples by determining the seroepidemiology of Brucellosis among livestock in 24 geographical areas in Greece using the Rose Bengal Test (RBT) and the complement fixation test (CFT) and further performing genetic analysis of Brucella spp. by species-specific real-time PCR and MLVA Brucella analysis. A total of 3086 serum samples from goats, sheep, and cattle showed positive results using the RBT and CFT, and only strongly positive samples (n = 800) were preserved in the Βlood Bank of the Veterinary Laboratory of Brucellosis. From these, 212 sera samples were randomly selected for molecular and genetic analysis. The results indicated that the incidence rate of Brucellosis is higher in cattle herds in comparison with other animal species. Overall, 48 samples tested positive by real-time PCR, of which forty-seven of them were B. abortus and one was B. melitensis. Genetic analysis of two B. abortus samples revealed a common pattern, indicating two Bruce04, two Bruce18, four Bruce07, two Bruce09, three Bruce16, and four Bruce30 for both samples, which, interestingly, were not identical with the known genotypes in the public MLVA Brucella database. Our findings substantiate that animal Brucellosis remains a health issue in Greece, with a stable but apparent incidence rate, and further investigation is needed to fully characterize the newly identified Brucella strains in Greece.

Applying a Fluorescence Polarization Assay for Detection of Brucellosis in Animals Using the Fluorescently Labeled Synthetic Oligosaccharides as Biosensing Tracer.

Brucellosis in animals is an infectious disease caused by bacteria of the genus Brucella. Known methods for diagnosing brucellosis face some challenges, due to the difficulties in isolating and standardizing the natural brucellosis antigen. In this work, we investigated the possibility of using the fluorescence polarization assay (FPA) with synthetic glycoconjugate biosensing tracers to detect antibodies against Brucella as a new methodology for diagnosing brucellosis. Based on the received results, the synthetic fluorescein-labeled trisaccharide tracer is most effective for Brucellosis detection. This tracer is structurally related to the immune determinant fragment of the Brucella LPS buildup of N-formyl-d-perosamine units, connected via α-(1→3)-linkage at the non-reducing end and α-(1→2)-linkage at the reducing end. The sensitivity and specificity in the case of the use of trisaccharide tracer 3b were 71% and 100% (Yuden's method) and 87% and 88% (Euclidean method), respectively, which is comparable with the diagnostic efficiency of traditionally used serological methods, such as the agglutination test (AT), complement fixation test (CFT), and Rose Bengal test (RBT). Given the known advantages of FPA (e.g., speed, compactness of the equipment, and standard reagents) and the increased specificity of the developed test system, it would be appropriate to consider its widespread use for the diagnosis of brucellosis in animals, including rapid testing in the field.

Obtaining and Characterization of Experimental Diagnostic Brucellosis Rabbit Sera for the Development of a National Standard for Brucellosis Serum

Relevance. The production of brucellosis diagnosticums is currently being established in the medical and veterinary practice of the Republic of Uzbekistan. However, due to the lack of a national standard serum in the country and the high cost of analog standard serums on the market, problems may arise in the production of diagnostic tools. In addition, the lack of national standard serums causes problems in quality control of commercial and local antigens, hinders the development of local drugs – brucellosis diagnostics, which reduces the effectiveness of serological diagnosis of brucellosis in humans and animals. Given the above, it becomes necessary to develop and produce a standard serum with a high level of safety, which will improve the methods of laboratory diagnosis of various forms of brucellosis in patients and animals to identify hidden foci of infection in Uzbekistan. The aim of the study is to obtain and characterize experimental standard diagnostic brucellosis rabbit sera for the development of national and industry standard serum with appropriate titers. Materials and Methods. The experiments were carried out in full compliance with the European Council Directive 2010/63/EU on compliance with ethical principles in working with laboratory animals and the experiments were carried out on the basis of the methodological manual «Methods and rules of work» approved by the Ministry of Health of the Republic of Uzbekistan in 2016 with laboratory animals in experimental microbiological and immunological studies. To obtain positive standard sera against brucellosis pathogens, 12 rabbits weighing from 2.2 kg to 4.6 kg, aged 6 to 12 months, were used. The animals were kept under identical conditions on a standard vivarium diet. For the experiment, animals adapted to the conditions of the study were selected and kept in quarantine for 21 days. The maintenance and conduct of the studies followed international rules for the humane treatment of animals. Results and Discussion. During the evaluation of the heterologic specificity of the obtained sera for 23 collection and hospital strains, a weakly positive response (+) was observed in 16.7% of cases. Rabbit sera positive for heterologic specificity were adsorbed on suspensions of inactivated microorganisms (Citrobacter spp. and Enterobacteriaceae spp.) - no significant drop in titer was observed, and cross-agglutination was eliminated. A significant increase in the total protein level was noted after the first and fourth immunizations. Before the first immunization, the total protein level was 59.57 g/l, and after the fourth immunization, it increased threefold to 157.17 g/l. There was also an increase in albumin and globulin (33.2 g/L and 26.37 g/L, respectively) to 79.37 g/L and 77.43 g/L, respectively. IgM levels remained almost unchanged, increasing 3.5-fold after the second immunization and decreasing to 0.24 mg/ml after the fourth immunization. An increase in the level of IgG after the second and third immunizations was revealed, followed by a decrease after the fourth immunization. Conclusion. The obtained national and industry standard diagnostic polyvalent brucellosis sera, as well as the developed methods of laboratory control of its quality and efficiency testing will make it possible to produce candidate standard serum samples for agglutination reaction test and detection of Brucella strains. The obtained standard serum samples will be further used for control of the produced immunobiological diagnostic preparations and provision of unified requirements to their quality. The goal was achieved – improving the serological diagnosis of brucellosis, laying the foundation for further study and standardization of serum.

Assessment of knowledge, attitude, and practice related to brucellosis among livestock farmers and meat handlers in Saudi Arabia.

Brucellosis is a bacterial zoonotic infection that is endemic in Saudi Arabia and associated with clinical and economic impacts. Several studies from countries endemic for brucellosis evaluated the knowledge and attitude of livestock farmers regarding brucellosis. However, no such study was conducted in Saudi Arabia. This study aimed to evaluate the knowledge, attitude, and practice of livestock farmers and meat handlers in Saudi Arabia. This was a cross-sectional questionnaire-based study, where participants were interviewed in-person in Arabic in livestock markets between September-December 2023. Convenient sampling was utilized. The questionnaire included basic demographics and questions to assess the knowledge, attitude, and practice toward personal protection and protection of the animals from brucellosis. The questionnaire was adapted from a previously validated survey and included 59 questions. Providing at least one correct answer to a certain question indicated a good knowledge about this item or a safe practice. The participants were divided into: farmers (shepherds working for the animal owners), commercial animal owners (those who rent a stockyard in the livestock market and employ farmers to sell their animals), and private animal owners (owners of private farms from which they sell their animals). 545 participants were interviewed (n = 291 farmers, n = 118 commercial animal owners, n = 113 private animal owners, and n = 23 animal slaughterhouse workers). >90% have heard of brucellosis. Lack of education and short experience (<5 years) of working with livestock were negatively associated with good knowledge of brucellosis symptoms and transmission (OR, 0.30; 95%CI, 0.10-0.94; p = 0.038 and OR, 0.23; 95%CI, 0.08-0.62; p = 0.004, respectively). Taking sick animals to the veterinarian was reported by 61.2%, whereas 36.4% follow safe practices when disposing of aborted fetuses. While 34% consume raw milk, only 10% consume rare/medium-rare meat. 51.2% acknowledged the need for more information on brucellosis. This study revealed the need to augment the knowledge of people working with animals, particularly those with no school education and those with short work experience, via providing educational visits or materials or through veterinarians. This should help them identify human and animal brucellosis symptoms and increase the knowledge on how to protect oneself and animals from this disease.

New synthesis of oligosaccharides modelling the M epitope of the Brucella O-polysaccharide.

Brucellosis is a dangerous zoonotic disease caused by bacteria of the genus Brucella. Diagnosis of brucellosis is based on the detection in animal and human sera of antibodies to the O-polysaccharide of Brucella lipopolysaccharide. The currently employed serodiagnosis of brucellosis relies on the use of the Brucella O-polysaccharide as a diagnostic antigen. However, the existence of bacterial species, which also express O-polysaccharides structurally similar to that of Brucella, may decrease the specificity of the brucellosis detection due to false-positive test results. It has been shown that the efficiency of the test can be significantly improved by using synthetic oligosaccharides that correspond to the so-called M epitope of the Brucella O-antigen. This epitope is characterized by an α-(1→3)-linkage between d-perosamine units and is unique to Brucella. Here we report on an efficient approach to the synthesis of oligosaccharides that model the M epitope of the Brucella O-polysaccharide. The approach is based on the use of the α-(1→3)-linked disaccharide thioglycoside as the key donor block. Its application allowed the straightforward assembly of a set of four protected oligosaccharides, which includes a disaccharide, two trisaccharides, and a tetrasaccharide, in five glycosylation steps. The synthesized oligosaccharides are planned to be used in the development of diagnostic tools for identifying brucellosis in humans and domestic animals, as well as a potential vaccine against it.

Production and evaluation of anti-BP26 monoclonal antibodies for the serological detection of animal brucellosis.

Brucella BP26 proves to be a highly immunogenic antigen with excellent specificity in brucellosis detection. In China, the authorized use of the Bp26-deleted vaccine M5ΔBP26 for preventing small ruminant brucellosis highlights the importance of developing accurate detection methods targeting BP26, particularly for the diagnosis of differentiation between infected and vaccinated animals (DIVA). Using the traditional mouse hybridoma technique, we successfully obtained 12 monoclonal antibodies (mAbs) targeting BP26. The efficacy of these mAbs in detecting various animal brucellosis cases using the competitive ELISA method was evaluated. Among them, only the E10 mAb exhibited significant efficiency, being inhibited by 100, 97.62, and 100% of brucellosis-positive sera from cattle, small ruminants, and canines, respectively. The E10-based competitive enzyme-linked immunosorbent assay (cELISA) outperformed the BP26-based indirect enzyme-linked immunosorbent assay (iELISA) in accuracy, particularly for cattle and small ruminant brucellosis, with cELISA sensitivity reaching 97.62% compared to 64.29% for iELISA for small ruminants. Although cELISA showed slightly lower specificity than iELISA, it still maintained high accuracy in canine brucellosis detection. The epitope of mAb E10 was identified in the amino acid sequence QPIYVYPDDKNNLKEPTITGY, suggesting its potential as a diagnostic antigen for brucellosis. In conclusion, the E10-based cELISA presents an effective means of detecting animal brucellosis, particularly significant for DIVA diagnosis in China, where the BP26-mutant vaccine is widely used.

Bioinformatics approach for structure modeling, vaccine design, and molecular docking of Brucella candidate proteins BvrR, OMP25, and OMP31

Brucellosis is a zoonotic disease with significant economic and healthcare costs. Despite the eradication efforts, the disease persists. Vaccines prevent disease in animals while antibiotics cure humans with limitations. This study aims to design vaccines and drugs for brucellosis in animals and humans, using protein modeling, epitope prediction, and molecular docking of the target proteins (BvrR, OMP25, and OMP31). Tertiary structure models of three target proteins were constructed and assessed using RMSD, TM-score, C-score, Z-score, and ERRAT. The best models selected from AlphaFold and I-TASSER due to their superior performance according to CASP 12 – CASP 15 were chosen for further analysis. The motif analysis of best models using MotifFinder revealed two, five, and five protein binding motifs, however, the Motif Scan identified seven, six, and eight Post-Translational Modification sites (PTMs) in the BvrR, OMP25, and OMP31 proteins, respectively. Dominant B cell epitopes were predicted at (44–63, 85–93, 126–137, 193–205, and 208–237), (26–46, 52–71, 98–114, 142–155, and 183–200), and (29–45, 58–82, 119–142, 177–198, and 222–251) for the three target proteins. Additionally, cytotoxic T lymphocyte epitopes were detected at (173–181, 189–197, and 202–210), (61–69, 91–99, 159–167, and 181–189), and (3–11, 24–32, 167–175, and 216–224), while T helper lymphocyte epitopes were displayed at (39–53, 57–65, 150–158, 163–171), (79–87, 95–108, 115–123, 128–142, and 189–197), and (39–47, 109–123, 216–224, and 245–253), for the respective target protein. Furthermore, structure-based virtual screening of the ZINC and DrugBank databases using the docking MOE program was followed by ADMET analysis. The best five compounds of the ZINC database revealed docking scores ranged from (− 16.8744 to − 15.1922), (− 16.0424 to − 14.1645), and (− 14.7566 to − 13.3222) for the BvrR, OMP25, and OMP31, respectively. These compounds had good ADMET parameters and no cytotoxicity, while DrugBank compounds didn't meet Lipinski's rule criteria. Therefore, the five selected compounds from the ZINC20 databases may fulfill the pharmacokinetics and could be considered lead molecules for potentially inhibiting Brucella’s proteins.

Специфическая профилактика бруцеллеза животных (современные проблемы)

Специфическая профилактика бруцеллеза животных (современные проблемы)

Brucellosis in camel, small ruminants, and Somali pastoralists in Eastern Ethiopia: a One Health approach.

Brucellosis is a neglected bacterial zoonotic disease with economic and public health importance in pastoral communities of sub-Saharan Africa. A cross-sectional study was conducted from December 2021 to April 2022, to estimate the prevalence and identify the associated risk factors causing brucellosis in animals and associated with occupational diseases in humans from three selected districts of "the Somali Pastoral region," Eastern Ethiopia. In this study, 1,000 serum samples were screened for anti-Brucella spp. antibodies using Rose Bengal Plate Test (RBPT) and further confirmed using a competitive enzyme-linked immunosorbent assay (cELISA). A structured questionnaire was used to collect the biodata of tested animals and animal attendants to test the association between explanatory and outcome variables. The overall animal level prevalence was 5% (95% CI, 6.1-7.2.0) in small ruminants, 2.9% (95% CI, 1.5-4.9) in camels, and 2.0% (95% CI, 0.2-3.7) in occupationally linked humans. Herd size and herd history of retained fetal membranes were risk factors associated with Brucella spp. seropositivity in animals (p < 0.05). Disposing of retained fetal membranes was significantly associated (p < 0.05) with Brucella spp. seropositivity in humans. Evidence of brucellosis in various livestock species and associated seropositivity in humans indicates the need for a coordinated One Health approach, considering sociocultural dynamics of pastoral communities in controlling brucellosis to safe guard public health and increase livestock productivity.

Exposure to Brucella Species, Coxiella burnetii, and Trichinella Species in Recently Imported Camels from Sudan to Egypt: Possible Threats to Animal and Human Health.

Brucellosis and coxiellosis/Q fever are bacterial infections caused by Brucella species and Coxiella burnetii, respectively; camels are highly susceptible to both pathogens. Trichinellosis is a parasitic infection caused by various Trichinella nematode species. Reportedly, camels are susceptible to experimental infection with Trichinella spp., but information on this potential host species is scarce. All three infections are of zoonotic nature and thus of great public health concern. The current study aimed to determine antibodies against the three pathogens in recently imported camels (n = 491) from Sudan at the two main ports for the entrance of camels into southern Egypt using commercial indirect ELISAs. Samples were collected in two sampling periods. The seropositivity rates of Brucella spp., C. burnetii, and Trichinella spp. were 3.5%, 4.3%, and 2.4%, respectively. Mixed seropositivity was found in 1% for Brucella spp. and C. burnetii. Marked differences were found between the two study sites and the two sampling periods for Brucella. A higher rate of seropositivity was recorded in the Red Sea/older samples that were collected between 2015 and 2016 (4.3%, 17/391; odds ratio = 9.4; p < 0.030) than in those collected in Aswan/recent samples that were collected between 2018 and 2021 (0/100). Concerning C. burnetii, samples collected during November and December 2015 had a significantly higher positivity rate than the other samples (13%, 13/100; OD = 4.8; p < 0.016). The same effect was observed for antibodies to Trichinella spp., with samples collected during November and December 2015 showing a higher positivity rate than the other samples (7%, 7/100; OD = 10.9; p < 0.001). This study provides valuable information on the seroprevalence of Brucella spp. and additional novel information on C. burnetii and Trichinella spp. in recently imported camels kept in quarantine before delivery to other Egyptian regions. This knowledge can be utilized to reduce health hazards and financial burdens attributable to brucellosis, Q fever, and trichinellosis in animals and humans in Egypt.

УСОВЕРШЕНСТВОВАНИЕ МЕТОДА ИММУНО–ПЦР В ЛАБОРАТОРНОЙ ДИАГНОСТИКЕ БРУЦЕЛЛЕЗА

Brucellosis of farm animals is one of the most difficult problems of veterinary science, practice and medicine, especially in the North Caucasus Federal District. Despite the control measures taken, this dangerous infectious disease is still widespread in economic entities of the country. Brucellosis in animals leads to serious losses in agriculture. To combat brucellosis in the North Caucasus Federal District, large-scale preventive measures are carried out, including vaccination of animals, food quality control and training of farmers and veterinary specialists. However, despite all efforts, brucellosis remains a significant problem and requires further research, development of new methods, their improvement to expand diagnostic capabilities, open new horizons in the study of the disease, optimize the work process and increase competitiveness in the scientific field. This is the only way to cope with this difficult task and ensure the safety of farm animals and people. This article substantiates the idea of improving the immuno-PCR method to expand diagnostic capabilities in the study of the disease, optimize the work process and increase competitiveness in the scientific field. During the study, positive and negative aspects of the use of immuno-PCR were identified in order to carry out a qualitative theoretical analysis, improve the strategy for examining foci in order to update the existing system of epidemiological control of brucellosis in time, we are faced with the task of revising and improving it. In general, expanding the diagnostic arsenal through new methods, their combination and improvement represents an important step in the study of brucellosis and other diseases, which allows us to obtain deeper and more accurate information, improve diagnostics and develop more effective methods for their control and prevention. Based on the powerful potential of iPCR, we can expect this method to become more widespread as it is improved and optimized.

Distribution, seroprevalence and risk factors for bovine brucellosis in Brazil: Official data, systematic review and meta-analysis

Bovine brucellosis is an endemic disease in Brazil, and evidence-based assessments of the available literature on its seroprevalence and risk factors are limited. The aim of this study was to systematically review and summarize studies related to seroprevalence and risk factors of bovine brucellosis in the entire Brazil, in addition to comparing published data with the most recent official reports. Articles available in scientific databases and published between October 2006 and October 2021 were evaluated. Forty-five publications were included in the meta-analysis on the seroprevalence of brucellosis and 29 publications in the review on risk factors. The largest number of publications was found for the State of Mato Grosso do Sul (n=4), and the highest and lowest seroprevalences were observed in Acre (11%; 95% CI: 8.0–14.0%) and in the Federal District (0.4%; 95% CI: 0.2–0.7%). The main risk factors were the purchase of animals for breeding, vaccination, the number of heifers (female ≥2 years), the presence of calving paddocks and the occurrence of abortions. The need for new official studies has been suggested to determine the true prevalence of bovine brucellosis in Brazil, supported by the National Program for the Control and Eradication of Animal Brucellosis and Tuberculosis.

Detectable immunological response in Brucella abortus s 19 vaccinated herds of cattle in vilachchiya veterinary range, north central province, Sri Lanka

Brucellosis a zoonotic disease caused by Gram-negative, intracellular, coccobacillus. Brucellosis, an economically important disease in livestock with zoonotic implications has been reported worldwide. It is transmitted via contaminated animals and their products. Clinical signs of animal brucellosis are abortions, reduced fertility, retention of fetal membranes, endometritis, and substantial decline in milk production over an animal’s lifespan. Orchitis, epididymitis and chronic hygroma had been reported in bulls. The current study focused on to studying the detectable immunological responses to Brucella S 19 live attenuated vaccine using serological tests in cattle of dry zone farms. A questionnaire-based survey was carried out in selected farms with and without a previous history of abortion respectively. Animals that were in 1st to 5th lactation in the 18 farms were selected for screening by Rose Bengal Plate Test (RBPT) and positives were confirmed by Complement Fixation Test (CFT). Three farms each with and without history of abortion were selected for Brucella S19 vaccination from the RBPT and CFT negative farms. Sixty non pregnant heifers over 6 months were selected, and 1ml each of S19 vaccine was given subcutaneously to 30 animals while 30 animals were kept as controls. The serum samples were collected and screened for Brucellosis by RBPT and CFT at 2, 4, 6, 8 and 10 weeks of post vaccination. The second week after the vaccination, 71.42% of the animals showed over 1:4 CFT levels while 28.57% of them did not show any antibody titres for Brucellosis in the Brucella endemic farms. In Brucella non-endemic farms, 68.75% of animals showed over 1:4 CFT titre at 2 weeks after the vaccination. However, 31.25% of animals did not show any CFT titre. No persistent and detectable antibody responses were found 10 weeks post vaccination in heifers of endemic and non-endemic herds. The titers of CFT indicate IgG or IgM humeral response, cell mediated immune response of live attenuated vaccine has not been monitored in the study. Thus, seropositivity after the 10 weeks post vaccination may indicate an infection of Brucellosis since no antibodies from vaccination were found. Further investigation with highly sensitive tests such as ELISA is recommended to have a better insight into this aspect in the future.

Applying the CRISPR/Cas9 for Treating Human and Animal Diseases – Comprehensive Review

Abstract Recently, genome editing tools have been extensively used in many biomedical sciences. The gene editing system is applied to modify the dnA sequences in the cellular system to comprehend their physiological response. A developing genome editing technology like clustered regularly short palindromic repeats (CRISPR) is widely used in medical sciences. CRISPR and CRISPR-associated protein 9 (CRISPR/Cas9) system is being exploited to edit any DNA mutations related to inherited ailments to investigate in animals (in vivo) and cell lines (in vitro). Remarkably, CRISPR/Cas9 could be employed to examine treatments of many human genetic diseases such as cystic fibrosis, tyrosinemia, phenylketonuria, muscular dystrophy, Parkinson’s disease, retinoschisis, hemophilia, β-thalassemia and atherosclerosis. Moreover, CRISPR/Cas9 was used for disease resistance such as tuberculosis, Johne’s diseases, chronic enteritis, and brucellosis in animals. Finally, this review discusses existing progress in treating hereditary diseases using CRISPR/Cas9 technology and the high points accompanying obstacles.

Brucella spp. are facultative anaerobic bacteria under denitrifying conditions.

Bacteria of the genus Brucella are responsible for human and animal brucellosis worldwide and comprise the well-characterized, so-called classical species belonging to the core clade, with a host range limited to mammals, and the novel atypical strains mainly considered as non-zoonotic to date, but able to colonize non-traditional hosts such as amphibians. Despite marked phenotypical differences between the two groups, Brucella species display a high degree of genome identity, including genes encoding reductases involved in the denitrification pathway, a respiratory process able to reduce nitrate to nitrogen under anoxic conditions. Brucella spp. have always been considered as strictly aerobic pathogens. Our study, however, describes for the first time that atypical Brucella spp. grow well under anoxia, but in a strictly nitrate-dependent manner, whereas classical species show a heterogeneous behavior. The rate of anaerobic growth of the atypical species Brucella microti was dependent on nitrate concentration, highlighting a more efficient denitrification pathway with rapid consumption of intermediate nitrite, whereas the classical species Brucella suis exhibited lower, nitrate concentration-independent growth coupled with nitrite accumulation. Transcriptional studies performed under anoxic conditions revealed a higher expression of genes participating in the last three denitrification steps in B. microti, as compared to B. suis, which correlates with the rapid and efficient consumption of nitrite in this atypical species and its rapid growth.IMPORTANCERespiration is a fundamental and complex process that bacteria use to produce energy. Despite aerobic respiration being the most common, some bacteria make use of a mode of respiration in the absence of oxygen, called anaerobic respiration, which can yield advantages in adaptation to various environmental conditions. Denitrification is part of this respiratory process ensuring higher respiratory flexibility under oxygen depletion. Here, we report for the first time the evidence of anaerobic growth of Brucella spp. under denitrifying conditions, which implies that this genus should be reconsidered as facultative anaerobic. Our study further describes that efficient denitrification is not equally found within the Brucella genus, with atypical species showing a greater ability to denitrify, correlated with higher expression of the genes involved, as compared to classical species.

Relief of brucellosis in an acute focus of returning infection with the use of a vaccine from the strain B. abortus 82

The purpose of the study was to study the features of the manifestation of the epizootic process during a returning infection in a fresh focus of brucellosis in cattle. The work was carried out on a dairy farm on a herd of cattle of a black-and-white breed, for a long time successful in brucellosis. The intensity of the epizootic process was assessed based on the results of serological studies of cattle of all sex and age groups. Immunological studies were carried out in accordance with regulatory documents on the diagnosis of brucellosis of animals. Standard diagnostic kits (S-) from commercial kits were used for serological studies in generally accepted reactions. Differentiation of positive and doubtful reactions was carried out using an experimental sample of an antigen made from brucella in R-form. A dynamic spread of brucellosis was noted during the six months with the 7% ill animals among all of the receptive cattle. Wellness measures were carried out with using the poorly agglutinogenic vaccine of strain Brucella abortus 82. The recovery period was 24 months. During of the trouble period 23% of infected animals were detected, including 19% of cows and 4% of young animals. The number of sick animals in a percent was estimated from the total numbers of the cattle in the farm. Based on the data obtained, it was established that under modern conditions, against the background of longterm well-being in a fresh focus of brucellosis, a dynamic spread of infection was noted with a wide coverage of susceptible livestock and a high intensity of the epizootic process.