scholarly journals A Novel Evidence of Immunological and Molecular Detection of Brucella Species in Camels

2021 ◽  
pp. 78-87
Author(s):  
Kavitha Manivannan ◽  
Malathi Ramasamy ◽  
Hanaa Ahmed
Keyword(s):  
Rose Bengal ◽  
Protein Gene ◽  
Ring Test ◽  
Pcr Analysis ◽  
Brucella Species ◽  
Rt Pcr ◽  
Dromedary Camels ◽  
Brucella Infection ◽  
Immunological Assays ◽  
Milk Ring Test

Objectives: Brucellosis is a zoonosis with severe complications for both humans and animals. In this work, we intended to examine the Brucella infection in dromedary camels in Qatar by using different analysis. Materials and Methods: A total of 203 samples of dromedary camels were randomly collected from the nearby farms in Qatar. Real-time PCR for the genus specific Brucella cell surface salt extractable bcsp31 kDa protein gene were performed on DNA extracted from camel samples. Rose Bengal and rivenol tests were performed to detect the Brucella species. The milk samples were collected from the camels and utilized for the milk ring test. Results: The outcomes of RT-PCR analysis illustrate the presence of Brucella spp. in 170 samples (83.74%) out of 203 samples. The findings of immunological assays also proved the presence of Brucella spp. such as Rose Bengal (67.14%), ELISA (71.42%), and precipitation assay (65.71) in both serum and blood samples of the dromedary camels, which were collected from the Qatar. Conclusions: In conclusion, it was clear that the incidence of the brucellosis in camels is significantly rising in Qatar region and there is a need to control the spread of the disease from camels to camels as well as from camels to humans.

scholarly journals Prevalence of Caprine brucellosis in Anhui province, China

2019 ◽  
Vol 12 (4) ◽  
pp. 558-564 ◽  
Author(s):  
Sajid Ur Rahman ◽  
Lei Zhu ◽  
Li Cao ◽  
Yafei Zhang ◽  
Xiaoyan Chu ◽  
...  
Keyword(s):  
Zoonotic Diseases ◽  
Anhui Province ◽  
Ring Test ◽  
Molecular Method ◽  
Young Males ◽  
Brucella Infection ◽  
Males And Females ◽  
Polymerase Chain ◽  
Serum Agglutination ◽  
Milk Ring Test

Background and Aim: Brucellosis is one of the most widespread zoonotic diseases globally. Studies indicated the existence of Brucella infection in goats in some province of China. Thus this study aimed to estimate the prevalence of brucellosis in goats of Anhui Province, China. Materials and Methods: Serum and milk samples obtained from goats in different regions of Anhui province were studied through rose Bengal plate test (RBPT), serum agglutination test (SAT), milk ring test (MRT), and polymerase chain reaction (PCR). Results: The investigation frequency of brucellosis using RBPT, SAT, MRT, and PCR methods was 3.9% (n=7), 4.45% (n=8), 11.67% (n=7), and 86.67% (n=156), respectively. The prevalence recorded for brucellosis in sex-wise animals as in females 5.55%, 6.67%, 11.67%, and 78.8% through above methods, while in males, it was 2.23% and 2.23% by RBPT and SAT. However, in age-wise animals, the results 6.36%, 7.27%, 11.67%, and 74.5% were perceived positive by RBPT, SAT, MRT, and PCR in adult females, respectively, but young males and females (up to 9 months) were considered free from brucellosis. Conclusion: These results show that prevalence of brucellosis was relatively higher in females than male's goats and SAT was relatively specific and accurate as compared to RBPT and MRT, but for diagnosis of brucellosis, molecular method (PCR) is recommended.

scholarly journals The use of the milk ring test and rose bengal test in brucellosis control and eradication in Nigeria

2008 ◽  
Vol 79 (3) ◽  
Author(s):  
S.I.B. Cadmus ◽  
H.K. Adesokan ◽  
J. Stack
Keyword(s):  
Rose Bengal ◽  
Screening Tests ◽  
Ring Test ◽  
Serum Samples ◽  
Chi Square ◽  
Southwestern Nigeria ◽  
Rose Bengal Test ◽  
The Difference ◽  
Control And Eradication ◽  
Milk Ring Test

In this study, milk and blood samples collected simultaneously from 532 trade cows to be slaughtered at Bodija abattoir, Ibadan (southwestern, Nigeria) were examined for antibodies to Brucella using the milk ring test (MRT) and the rose bengal test (RBT). Overall, 18.61%of the milk samples were positive according to the MRT, while 9.77 % of the serum samples were positive according to the RBT. The difference was highly significant (Chi-square value 16.33; P<0.05); only 32 (6.02 %) of the samples were positive for both tests. The Red Bororo breed of cattle and the White Fulani had the highest positive rates, namely 20.93 % and 11.69%for the MRT and RBT respectively.No conclusion can be drawn about sensitivity because we do not know the true status of the animals tested. It is, however, obvious that although the MRT and RBT are 1st-line screening tests for brucellosis in cows in some countries, their lack of specificity is of concern. Therefore, the requirement for other confirmatory tests that are more specific should be considered for control and eradication of the disease, especially in Nigeria.

scholarly journals Identification of a novel lineage of Crimean–Congo haemorrhagic fever virus in dromedary camels, United Arab Emirates

2020 ◽  
Author(s):  
Abdelmalik I. Khalafalla ◽  
Yan Li ◽  
Anna Uehara ◽  
Nasareldien A. Hussein ◽  
Jing Zhang ◽  
...  
Keyword(s):  
United Arab Emirates ◽  
Asymptomatic Infection ◽  
Fever Virus ◽  
Pcr Analysis ◽  
Molecular Evidence ◽  
Haemorrhagic Fever ◽  
Rt Pcr ◽  
Dromedary Camels ◽  
High Fatality Rate ◽  
Crimean Congo Haemorrhagic Fever

Crimean–Congo haemorrhagic fever virus (CCHFV) is a tick-borne virus causing Crimean–Congo haemorrhagic fever (CCHF), a disease reported to have a high fatality rate in numerous countries. The virus is geographically widespread due to its vector, and numerous wild and domestic animals can develop asymptomatic infection. Serological and limited molecular evidence of CCHFV has previously been reported in Camelus dromedarius (the dromedary, or one-humped camel) in the United Arab Emirates (UAE). In this study, 238 camel samples were screened for CCHFV RNA where 16 camel samples were positive for CCHFV by RT-PCR. Analysis of full-length CCHFV genome sequences revealed a novel lineage in camels from the UAE, and potential reassortment of the M segment of the genome.

scholarly journals BACTERIOLOGICAL & SEROLOGICAL STUDY OF BRUCELLOSIS IN BUFFALO IN BAGHDAD

2006 ◽  
Vol 30 (2) ◽  
pp. 23-30
Author(s):  
S. A. Al-Bayatti
Keyword(s):  
Rose Bengal ◽  
Brucella Abortus ◽  
Agglutination Test ◽  
Ring Test ◽  
Serological Study ◽  
Isolation And Identification ◽  
Milk Samples ◽  
Milk Ring Test

To study the prevalence of brucellosis among buffalo in Baghdad, 144serum samples were collected and tested serologically using Rose Bengal PlateTest (RBPT) and Tube Agglutination Test (TAT), also 44 milk samples testedby Milk Ring Test (MRT). For isolation and identification of Brucella speciesand behavior, 15 buffalo aborted fetuses were used. And for evaluation of thehealth hazzard caused by the consumption of the locally manufactured creamfrom the milk of buffalo, 76 cream samples were used for cultivation.The results of the RBPT, TAT and MRT revealed the prevalence of thedisease in the percentages (38.19%, 29.86% and 20.45%) respectively.It was found that Brucella abortus was the main cause of abortion in thebuffalo (53.33%). There biotypes were the 8th and 3rd.The cream samples manufactured from the milk of buffalo doesn’t showhealth hazzard as a cause of brucellosis in human, because it didn’t revealed anyBrucella isolates in all samples used.

scholarly journals Dose-dependent Inhibition of Gynecophoral Canal Protein Gene Expression in Vitro in the Schistosome (Schistosoma japonicum) by RNA Interference

2005 ◽  
Vol 37 (6) ◽  
pp. 386-390 ◽  
Author(s):  
Guo-Feng Cheng ◽  
Jiao-Jiao Lin ◽  
Yi Shi ◽  
You-Xin Jin ◽  
Zhi-Qiang Fu ◽  
...  
Keyword(s):  
Schistosoma Japonicum ◽  
Protein Gene ◽  
Female Worm ◽  
Transcript Level ◽  
Pcr Analysis ◽  
Rt Pcr ◽  
Male Worm ◽  
Dependent Inhibition ◽  
Dose Dependent

Abstract The gynecophoral canal protein gene SjGCP of Schistosoma japonicum that is necessary for the pairing between the male and female worms is specifically expressed in the adult male worm. This protein is widely distributed in the adult female worm after pairing. Reverse transcription-polymerase chain reaction (RT-PCR) and immunofluorescence were employed to analyze the relationship between the RNAi effect and dsRNA dosage in the parasites. The results revealed that the inhibition of SjGCP expression by siRNA is dose-dependent. RT-PCR analysis showed that the SjGCP transcript level was reduced by 75% when 100 nM dsRNA was applied.

scholarly journals Phenotypical and Functional Characteristics of Human Regulatory T Cells during Ex Vivo Maturation from CD4+ T Lymphocytes

2021 ◽  
Vol 11 (13) ◽  
pp. 5776
Author(s):  
Varvara G. Blinova ◽  
Natalia S. Novachly ◽  
Sofya N. Gippius ◽  
Abdullah Hilal ◽  
Yulia A. Gladilina ◽  
...  
Keyword(s):  
T Cells ◽  
Regulatory T Cells ◽  
Ex Vivo ◽  
Pcr Analysis ◽  
Mrna Levels ◽  
Rt Pcr ◽  
Inflammatory Reactions ◽  
Effector Cells ◽  
Cycle Regulation ◽  
Further Development

Regulatory T cells (Tregs) participate in the negative regulation of inflammatory reactions by suppressing effector cells. In a number of autoimmune disorders, the suppressive function and/or the number of Tregs is compromised. The lack of active functioning Tregs can be restored with adoptive transfer of expanded ex vivo autologous Tregs. In our study, we traced the differentiation and maturation of Tregs CD4+CD25+FoxP3+CD127low over 7 days of cultivation from initial CD4+ T cells under ex vivo conditions. The resulting ex vivo expanded cell population (eTregs) demonstrated the immune profile of Tregs with an increased capacity to suppress the proliferation of target effector cells. The expression of the FoxP3 gene was upregulated within the time of expansion and was associated with gradual demethylation in the promotor region of the T cell-specific demethylation region. Real-time RT-PCR analysis revealed changes in the expression profile of genes involved in cell cycle regulation. In addition to FOXP3, the cells displayed elevated mRNA levels of Ikaros zinc finger transcription factors and the main telomerase catalytic subunit hTERT. Alternative splicing of FoxP3, hTERT and IKZF family members was demonstrated to be involved in eTreg maturation. Our data indicate that expanded ex vivo eTregs develop a Treg-specific phenotype and functional suppressive activity. We suggest that eTregs are not just expanded but transformed cells with enhanced capacities of immune suppression. Our findings may influence further development of cell immunosuppressive therapy based on regulatory T cells.

scholarly journals Role of Innate Inflammation in the Regulation of Tissue Remodeling during Tooth Eruption

2021 ◽  
Vol 9 (1) ◽  
pp. 7
Author(s):  
Yusuke Makino ◽  
Kaoru Fujikawa ◽  
Miwako Matsuki-Fukushima ◽  
Satoshi Inoue ◽  
Masanori Nakamura
Keyword(s):  
Bacterial Infections ◽  
Tooth Movement ◽  
Tooth Eruption ◽  
Intercellular Adhesion Molecule ◽  
Enamel Organ ◽  
Pcr Analysis ◽  
Maturation Stage ◽  
Oral Epithelium ◽  
Rt Pcr ◽  
Inflammatory Reactions

Tooth eruption is characterized by a coordinated complex cascade of cellular and molecular events that promote tooth movement through the eruptive pathway. During tooth eruption, the stratum intermedium structurally changes to the papillary layer with tooth organ development. We previously reported intercellular adhesion molecule-1 (ICAM-1) expression on the papillary layer, which is the origin of the ICAM-1-positive junctional epithelium. ICAM-1 expression is induced by proinflammatory cytokines, including interleukin-1 and tumor necrosis factor. Inflammatory reactions induce tissue degradation. Therefore, this study aimed to examine whether inflammatory reactions are involved in tooth eruption. Reverse transcription-polymerase chain reaction (RT-PCR) analysis revealed sequential expression of hypoxia-induced factor-1α, interleukin-1β, and chemotactic factors, including keratinocyte-derived chemokine (KC) and macrophage inflammatory protein-2 (MIP-2), during tooth eruption. Consistent with the RT-PCR results, immunohistochemical analysis revealed KC and MIP-2 expression in the papillary layer cells of the enamel organ from the ameloblast maturation stage. Moreover, there was massive macrophage and neutrophil infiltration in the connective tissue between the tooth organ and oral epithelium during tooth eruption. These findings suggest that inflammatory reactions might be involved in the degradation of tissue overlying the tooth organ. Further, these reactions might be induced by hypoxia in the tissue overlying the tooth organ, which results from decreased capillaries in the tissue. Our findings indicate that bacterial infections are not associated with the eruption process. Therefore, tooth eruption might be regulated by innate inflammatory mechanisms.

scholarly journals RT-PCR analysis of Deformed wing virus in honeybees (Apis mellifera) and mites (Varroa destructor)

2005 ◽  
Vol 86 (12) ◽  
pp. 3419-3424 ◽  
Author(s):  
Constanze Yue ◽  
Elke Genersch
Keyword(s):  
Varroa Destructor ◽  
Pcr Analysis ◽  
Body Parts ◽  
Rt Pcr ◽  
Larval Food ◽  
Viral Pathogen ◽  
Deformed Wing Virus ◽  
Significant Difference ◽  
Almost All ◽  
Viral Sequences

Deformed wing virus (DWV) is a honeybee viral pathogen either persisting as an inapparent infection or resulting in wing deformity. The occurrence of deformity is associated with the transmission of DWV through Varroa destructor during pupal stages. Such infections with DWV add to the pathology of V. destructor and play a major role in colony collapse in the course of varroosis. Using a recently developed RT-PCR protocol for the detection of DWV, individual bees and mites originating from hives differing in Varroa infestation levels and the occurrence of crippled bees were analysed. It was found that 100 % of both crippled and asymptomatic bees were positive for DWV. However, a significant difference in the spatial distribution of DWV between asymptomatic and crippled bees could be demonstrated: when analysing head, thorax and abdomen of crippled bees, all body parts were always strongly positive for viral sequences. In contrast, for asymptomatic bees viral sequences could be detected in RNA extracted from the thorax and/or abdomen but never in RNA extracted from the head. DWV replication was demonstrated in almost all DWV-positive body parts of infected bees. Analysing individual mites for the presence of DWV revealed that the percentage of DWV-positive mites differed between mite populations. In addition, it was demonstrated that DWV was able to replicate in some but not all mites. Interestingly, virus replication in mites was correlated with wing deformity. DWV was also detected in the larval food, implicating that in addition to transmission by V. destructor DWV is also transmitted by feeding.

scholarly journals Reproductive failure in mice lacking inter-alpha-trypsin inhibitor (ITI) – ITI target genes in mouse ovary identified by microarray analysis

2004 ◽  
Vol 183 (1) ◽  
pp. 29-38 ◽  
Author(s):  
Mika Suzuki ◽  
Hiroshi Kobayashi ◽  
Yoshiko Tanaka ◽  
Naohiro Kanayama ◽  
Toshihiko Terao
Keyword(s):  
Real Time ◽  
Trypsin Inhibitor ◽  
Target Genes ◽  
Reproductive Function ◽  
Pcr Analysis ◽  
Rt Pcr ◽  
Reproductive Process ◽  
Female Mice ◽  
Retinoid Metabolism ◽  
Regulatory Effects

Bikunin, a Kunitz-type protease inhibitor, is found in blood and urine. It has been established by two laboratories independently that the bikunin knockout female mice display a severe reduction in fertility: the cumulus oophorus has a defect in forming the extracellular hyaluronan-rich matrix during expansion. Proteins of the inter-alpha-trypsin inhibitor (ITI) family are eliminated in mice in which the bikunin gene has been inactivated, since bikunin is essential for their biosynthesis. Proteins of the ITI family may contribute to the microenvironment in which ovulation takes place. It is not clear, however, whether a single mechanism affects the reproductive function including ovulation. For identifying the full repertoire of the ITI deficiency-related genes, a cDNA microarray hybridization screening was conducted using mRNA from ovaries of wild-type or bik−/− female mice. A number of genes were identified and their regulation was confirmed by real-time RT-PCR analysis. Our screen identified that 29 (0.7%) and 5 genes (0.1%) of the genes assayed were, respectively, up- and down-regulated twofold or more. The identified genes can be classified into distinct subsets. These include stress-related, apoptosis-related, proteases, signaling molecules, aging-related, cytokines, hyaluronan metabolism and signaling, reactive oxygen species-related, and retinoid metabolism, which have previously been implicated in enhancing follicle development and/or ovulation. Real-time RT-PCR analysis confirmed that these genes were up- and down-regulated two- to tenfold by bikunin knockout. These studies demonstrate that proteins of the ITI family may exert potent regulatory effects on a major physiological reproductive process, ovulation.

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