Standard Agglutination Research Articles

Brucellosis Coinfecting with Liver Hydatid Cyst: Case Report and Literature Review of Zoonoses Coinfection of Human Brucellosis.

Background: Brucellosis is a major global public health problem. Brucellosis usually coinfects with zoonoses because of their similar reservoir and infection routes. Materials and Methods: Here, we present a patient who was diagnosed with coinfecting with liver cyst and brucellosis. Results: A 57-year-old female farmer was hospitalized with right hip joint pain approximately 4 weeks earlier. The patient reported a history of hydatid cysts and a family history of brucellosis. Abdominal computerized tomography and ultrasonography revealed a liver hydatid cyst. The standard agglutination test titer of Wright's test was 1:100. Computerized tomography and magnetic resonance imaging revealed brucellosis hip arthritis. The patient was treated with a combination of doxycycline, rifampicin, and cefotaxime. The symptoms immediately improved. The patient was well and asymptomatic during the 6-month follow-up. We reviewed the literature on other zoonoses superinfection with brucellosis. Conclusion: Brucellosis coinfects with multiple zoonosis organisms and poses a serious health threat to humans. Awareness of possible brucellosis coinfection with other zoonoses, especially professionally exposed persons in endemic regions, is warranted.

Development and Evaluation of a Noninvasive Microfluidic-Based Paper Analytical Device for Leptospirosis Diagnosis.

Leptospirosis is a re-emerging infectious disease that presents a diagnostic enigma for clinicians with frequent misdiagnosis due to lack of rapid and accurate diagnostic tests, as the current methods are encumbered by inherent limitations. The development of a diagnostic sensor with a sample-in-result-out capability is pivotal for prompt diagnosis. Herein, we developed a microfluidic paper-based analytical device (spin-μPAD) featuring a sample-in-result-out fashion for the detection of Leptospira specific urinary biomarker, sph2 sphingomyelinase, crucial for noninvasive point-of-care testing. Fabrication of paper devices involved precise photolithography techniques, ensuring a high degree of reproducibility and replicability. By optimizing the device's configuration and protein components, a remarkable sensitivity and specificity was achieved for detecting leptospiral sph2 in urine, even at low concentrations down to 1.5 fg/mL, with an assay time of 15 min. Further, the spin-μPAD was validated with 20 clinical samples, suspected of leptospirosis including other febrile illnesses, and compared with gold standard microscopic agglutination test, culture, Lepto IgM ELISA, darkfield microscopy, and Leptocheck WB spot test. In contrast to commercial diagnostic tools, the spin-μPAD was noninvasive, rapid, easy to use, specific, sensitive, and cost-effective. The results highlight the potential of this innovative spin-μPAD for an efficient and dependable approach to noninvasive leptospirosis diagnosis, addressing critical needs in the realms of public health and clinical settings.

Brucella Seropositivity at a Tertiary Care Hospital in Chandigarh.

There are reports that the neglected zoonotic tropical disease brucellosis is reemerging today. Serological tests are being widely used in the diagnosis of brucellosis. In the present study, we performed a standard agglutination test (SAT) on 1348 suspected cases of brucellosis during the period of four years from April 2018 to March 2022. We noticed an increase in seropositivity from 2.6% in the year 2018-19 to 7.4% in the year 2021-22. We also noticed a spike in seropositivity in the years 2019-20 (12.5%). Our study shows the recent trend in seropositivity of the disease in Chandigarh and, hence, can be a meaningful addition to the existing serological diagnostic data related to brucellosis.

Improved Antibody Detection for Canine Leptospirosis: ELISAs Modified Using Local Leptospiral Serovar Isolates from Asymptomatic Dogs.

Leptospirosis is a zoonotic disease of significant concern for human and animal health, with domestic animals, including dogs, acting as reservoirs for human infection. Serology is widely used for leptospirosis diagnosis, even though the standard microscopic agglutination test (MAT) using a panel of serovars lacks specificity and can lead to detection limitations in certain regions. In this study, we aimed to develop an antibody detection tool for dogs using an indirect enzyme-linked immunosorbent assay (ELISA) with a set of local serovar isolates, including Paidjan, Dadas, and Mini, to enhance the accuracy of leptospirosis surveillance in our region. The specificity and sensitivity of various antigen preparations, namely leptospiral whole-cell protein (WCP), total membrane protein (TMP), and outer membrane protein (OMP), were assessed using sera from infected and non-infected dogs, as well as negative puppy sera. Leptospirosis diagnosis was supported using a genus-specific nested polymerase chain reaction test on all collected sera. Protein preparations were validated using SDS-PAGE and Western blotting analysis. In the results, the standard MAT failed to detect antibodies in any of the dogs confirmed as being infected using PCR and isolation, highlighting its limitations. In contrast, the OMP-based ELISAs using local isolates of Leptospira serovars gave positive results with sera from all infected dogs, and negative results with sera from all dogs from non-endemic areas. IgG titres of infected and unvaccinated dogs from endemically affected areas were significantly higher than those in non-endemic regions. Using the OMP-based IgG/ELISAs with the local serovar Dadas resulted in higher specificity and lower sensitivity than when using the WCP- and TMP-based IgG/ELISAs. Agreement analysis revealed fair and moderate concordance between OMP-based IgG/ELISAs and PCR results, whereas slight and fair agreement was observed between OMP-based ELISAs and the MAT. Overall, the modified OMP-based IgG/ELISAs, utilising relevant local serovar isolates from dogs, demonstrated improved accuracy in detecting leptospirosis in the study area, overcoming the limitations of the MAT. This study highlights the importance of identifying and incorporating these local circulating serovar isolates into serological techniques for leptospirosis diagnosis and surveillance.

Distribution of ABO and Rhesus (D) blood group antigens among the Indian Kami population from the Sub-Himalayan region of West Bengal and Sikkim, India

Abstract BACKGROUND: Limited information is available regarding the distribution of ABO and Rhesus (Rh) blood groups among the populations from the sub-Himalayan regions. The present study was undertaken to understand the phenotypic, genotypic, and allelic frequencies of ABO and Rh antigens among the Kami population. METHODS: A total of 1420 Kami individuals were included in the study from the sub-Himalayan region of West Bengal and Sikkim. The ABO and RhD blood groups were determined among 158 individuals by standard slide agglutination method, and from 1262 individuals blood group, data were collected who had undergone routine blood group tests in diagnostic laboratories. RESULTS: The frequency of blood group O (31.06%) was found to be the highest followed by B (29.15%), A (27.54%), and AB (12.25%). A total of 97.89% individuals were Rh (D) positive and 2.11% were Rh (D) negative. The allele frequencies of ABO were found to be 0.200 for IA (p), 0.243 for IB (q), and 0.557 for IO (r). For Rh (D), the calculated gene frequencies were 0.855 for Rh positive (ID) and 0.145 for Rh negative (Id). Among the subcaste of Kami population, a significantly high frequency of blood group A was observed among Baraily and Lamgade, O among Sashankar, Dural, Diyali, and Dutraj, and B among Ramudamu, Koirala, and Silal. CONCLUSION: The study is the first of its kind to comprehensively describe the ABO blood group distribution among the Kami population which shows the same trend as the general Indian population.

Characterization of Brucella spp. circulating in industrial dairy cattle farms in Iran: a field study 2016 - 2023.

Bovine brucellosis, an infectious disease transmitted by Brucella melitensis and Brucella abortus, presents a significant zoonotic risk for agricultural economics and animal health. The primary objective of this study was to present a comprehensive understanding of the prevalence and features of Brucella strains within the industrial dairy farming sector in Iran. Rose Bengal plate test, standard agglutination test, and indirect enzyme linked immunosorbent assay tests were used to confirm all seropositive animals. A total number of 1,311 bovine samples from seropositive animals including were collected from 224 farms in 21 provinces of different regions of Iran and examined. The discovered Brucella isolates were phenotyped and molecularly characterized. The isolates were all B. abortus or B. melitensis. Bacteria analysis revealed that 70.53% of seropositive farms were tested positive for Brucella strains, predominantly B. melitensis biovar 1 (43.42%) and B. abortus biovar 3 (27.11%). Geographical distribution revealed that B. melitensis biovar 1 was the most common in dairy cow farms (16 provinces), followed by B. abortus biovar 3 (six provinces). Also, the prevalence of B. melitensis biovar 2, B. melitensis biovar 3, B. abortus biovar 1, B. abortus biovar 2 and RB51 vaccine were restricted to certain provinces. AMOS (abortus melitensis ovis suis)-polymerase chain reaction and Bruce-ladder PCR confirmed species identification. These results highlighted the complexity of bovine brucellosis in Iran and illustrated that B. melitensis was spread from small ruminants to cattle. This study provided important epidemiological insights for targeting future brucellosis control programs in the Iranian dairy farms.

The outbreak of migratory goat's brucellosis in the Swat ecosystem of Khyber Pakhtunkhwa.

Brucellosis is a major threat to public health especially in developing countries including Pakistan. This study reveals the characterisation ofBrucellaspecies affecting humans and goats in the Swat region of Khyber Pakhtunkhwa, Pakistan. Blood samples were collected from shepherds and goats and analysed by Rose Bengal precipitation test (RBPT), standard plate agglutination test (SPAT), polymerase chain reaction (PCR) and Sanger sequencing of 16S rRNA gene. The findings of the study indicated 24% (36/150) and 11.3% (17/150) positivity forBrucella abortusandBrucella melitensis,respectively, in human samples. In samples of goats, 26.66% (40/150) were positive forB. abortusand 16.66% (25/150) samples were positiveB. melitensisby SPAT. The species-specific PCR confirmedB. abortusin 24% (36/150) of human samples and 26.66% (17/150) of goat samples by targeting the IS711 locus. The remaining seropositive samples were confirmed asB. melitensisusing IS711 M species-specific primer. The sequences of the amplified fragments of the 16S rRNA gene were blasted, and phylogenetic analysis revealed thatBrucellaspecies circulating in the Swat district were closely related toB. melitensisandB. abortusreported from India, China, Philippines, and the United States (US) showing the existence of the possible epidemiological linkage among theBrucellaspecies. This study concluded that there was a higher prevalence ofB. abortus(26.6%) in humans and goats compared toB. melitensis(16.6%). These results revealed that theBrucellaspecies were circulating in both humans and goats in the study areas. The findings of the study concluded thatB. abortusandB. melitensiswere circulating in goats and shepherds with a higher prevalence ofB. abortusthanB. melitensis. Furthermore, theBrucellaspecies identified in Swat were phylogenetically related to theBrucellaspecies reported from India, China, Philippines and the US.Contribution:The proposed study covers the scope of the journal. The species of the genusBrucellaaffect both animals and shepherds. This study investigates the seroprevalence of brucellosis in shepherds and goats in different geographical areas in the Swat district. The phylogenetic analysis of theBrucellaspp. identified in Swat showed close relationships to theBrucellaspecies reported in India, China, Philippines and the US, which shows the possible epidemiological linkages between theBrucellaspp.

Seroprevalence of Chlamydia abortus and Brucella spp. and risk factors for Chlamydia abortus in pigs from China

Chlamydiosis and brucellosis induced abortions have resulted in significant economic losses in the global livestock industry. Although there have been numerous reports on these two diseases in ruminants in China, limited information is available regarding the prevalence of Chlamydia abortus (C. abortus) and Brucella spp. infection in pigs. This study aimed to investigate the prevalence of C. abortus and Brucella spp. infections in pig serum using serology and to identify potential risk factors. In total, 2816 serum samples were collected from 12 provinces in China. The presence of C. abortus antibodies was determined using an enzyme-linked immunosorbent assay (ELISA), while the presence of Brucella spp. antibodies was examined using the Rose Bengal Plate Test (RBPT) and the Standard Agglutination Test (SAT). The seroprevalences of C. abortus and Brucella spp. were 8.38 % (236/2816) and 0.11 % (3/2816), respectively. Geographical location, season, and age were found to be risk factors associated with C. abortus infection in pig herds in China (p<0.01), and the seropositive rate for C. abortus in sow herds was strongly associated with the occurrence of abortion (p<0.01). Overall, in China, pigs exhibit a higher seroprevalence of C. abortus, whereas the prevalence of Brucella is limited. This study represents the first comprehensive survey of C. abortus and Brucella spp. in pig herds in China that established potential risk factors and provided data for the prevention and control of intraspecies and interspecies transmission of C. abortus to humans.

Pathogenesis, clinical manifestations, treatment and prevention of brucellosis in humans and animals

Brucella genus is Gram-negative coccobacilli that cause a zoonotic disease called brucellosis in humans and animals. 12 species have been identified for Brucella, among which four species, Brucella melitensis, Brucella suis, Brucella abortus, and Brucella canis, commonly cause disease in humans and animals. Brucellosis is transmissible and there are different types of transmission such as animal-to-human (dairy and meat consumption), human-to-human (placenta, breastfeeding, sexual intercourse, blood transfusion, and bone marrow transplantation), and animal-to-animal (aerosol-containing Brucella, placenta and sexual intercourse). Due to having virulence factors such as type IV secretion system (T4SS), superoxide dismutase enzyme, BvrS/BvrR two-component system, LPS with O chain, and cyclic beta-glucan, Brucella can survive and replicate intracellularly and cause disease. Fever, excessive sweating, malaise, myalgia, arthralgia, loss of appetite, weight loss, hepatosplenomegaly, spondylitis, and arthritis are the clinical manifestations of brucellosis in humans. In animals, complications such as abortion, weak calf birth, reduced fertility, endometritis, reduced milk production, orchitis, epididymis, and hygroma may be observed. To diagnose brucellosis, a direct method such as culture or indirect and serological methods such as the Rose Bengal test (RBT), tube standard agglutination test (SAT), 2-mercaptoethanol test, Coombs test, and ELISA are used. According to the condition of the patient, the combination of doxycycline with streptomycin, gentamicin, or rifampin is used to treat brucellosis. While animals with brucellosis are not treated and are slaughtered. No vaccine against brucellosis has been approved for humans yet. But for animals, S19, Rev.1, S2, RB51, and SR82 vaccines are used for prevention. Development and approval of brucellosis vaccine for humans, wider use of animal vaccines, and timely diagnosis of suspected cases will help to better control brucellosis and prevent economic consequences.

The correlation between Blood group type and Diabetes Mellitus Type II

Background and objective: Type 2 diabetes mellitus is typically a multifactorial disorder involving genetic and environmental factors to variable extents. The aim of this research was to find out if there is a relationship between the “ABO” and “Rhesus” blood groups and type 2 diabetes mellitus. Methods: A case-control study was carried out at Azadi Teaching Hospital in Duhok city for six months, from 1st July 2019 to 1st January 2020. The study included 800 individuals including 400 randomly selected diabetic patients and 400 non-diabetic adults of both genders. Standard slide agglutination method at room temperature was performed to determine the ABO and Rh blood groups. Results: Out of 400 patients, 185(46.25%) were male and 215(53.75%) were female. It was found that out of 400 randomly selected patients, majority had O blood group 193(48.25%), 109(27.25%) had A, 65(16.25%) had B, 33(8.25%) had AB group. A higher frequency of O blood group was encountered among the diabetics comparing to the control (48.25% vs. 37.75%). On applying the Chi-square test, the blood groups O and type 2 diabetes mellitus were found to have a statistically significant relationship (Chi-square value - 8.14, P &lt;0.005) and no statistically significant association between the Rh group and type 2 DM was observed (P &gt;0.05). Conclusion: According to this study, people with the O blood group are at a greater risk of having type 2 diabetes mellitus, but there was no discrepancy in type 2 diabetes mellitus risk between Rhesus positive and negative classes. The level of blood groups A among diabetic patients is also concluded to be substantially lower.

Evaluating the efficacy of serological testing of clinical specimens collected from patients with suspected brucellosis.

This study aims to evaluate the efficacy of the standard agglutination test (SAT), the Brucellacapt test and enzyme-linked immunosorbent assay (ELISA) in clinical specimens collected from patients with suspected brucellosis. A prospective study was conducted from December 2020 to December 2021. Brucellosis was diagnosed on the basis of clinical evidence, and confirmed by isolation of Brucella or a four-fold rise in SAT titer. All samples were tested by the SAT, ELISA and the Brucellacapt test. Titers ≥1:100 were considered as SAT positive; ELISA was considered positive when an index greater than 11 was detected, while titers ≥1/160 indicated positivity on the Brucellacapt test. The specificity, sensitivity, and positive (PPVs) and negative predictive values (NPVs) of the three different methods were calculated. A total of 149 samples were collected from patients with suspected brucellosis. The sensitivities for the SAT, IgG, and IgM detection were 74.42%, 88.37% and 74.42%, respectively. The specificities were 95.24%, 93.65%, and 88.89%, respectively. The simultaneous measurement of IgG and IgM improved the sensitivity (98.84%) but reduced the specificity (84.13%) compared to each antibody test separately. The Brucellacapt test had excellent specificity (100%) and a high PPV (100%); however, the sensitivity and NPV were 88.37% and 86.30%, respectively. The combination of IgG detection by ELISA and the Brucellacapt test had excellent diagnostic performance, with 98.84% sensitivity and 93.65% specificity. This study showed that the simultaneous performance of IgG detection by ELISA and the Brucellacapt test has the potential to overcome the current limitations of detection.

Validation of a flow-cytometry-based red blood cell antigen phenotyping method.

Current manual and automated phenotyping methods are based on visual detection of the antigen-antibody interaction. This approach has several limitations including the use of large volumes of patient and reagent red blood cells (RBCs) and antisera to produce a visually detectable reaction. We sought to determine whether the flow cytometry could be developed and validated to perform RBC phenotyping to enable a high-throughput method of phenotyping using comparatively miniscule reagent volumes via fluorescence-based detection of antibody binding. RBC phenotyping by flow cytometry was performed using monoclonal direct typing antisera (human IgM): anti-C, -E, -c, -e, -K, -Jka , -Jkb and indirect typing antisera (human IgG): anti-k, -Fya , -Fyb , -S, -s that are commercially available and currently utilized in our blood transfusion services (BTS) for agglutination-based phenotyping assays. Seventy samples were tested using both flow-cytometry-based-phenotyping and a manual tube standard agglutination assay. For all the antigens tested, 100% concordance was achieved. The flow-cytometry-based method used minimal reagent volume (0.5-1μl per antigen) compared with the volumes required for manual tube standard agglutination (50μl per antigen) CONCLUSION: This study demonstrates the successful validation of flow-cytometry-based RBC phenotyping. Flow cytometry offers many benefits compared to common conventional RBC phenotyping methods including high degrees of automation, quantitative assessment with automated interpretation of results and extremely low volumes of reagents. This method could be used for high-throughput, low-cost phenotyping for both blood suppliers and hospital BTS.

Brucella cultures characteristics, clinical characteristics, and infection biomarkers of human Brucellosis

ObjectiveAim to investigate the brucella culture characteristics, diagnosis methods, and clinical characteristics, to provide the laboratory with diagnostic methods and prevention and treatment for brucellosis. MethodsData of 328 cases of brucellosis from 2012 to 2022 was analyzed, retrospectively. The bacterial culture characteristics, the clinical diagnostic methods, and the complications were analyzed respectively. The infection biomarkers of the brucellosis were analyzed by Receiver operating characteristic curve ROC. ResultsAmong the 328 brucellosis, 78.96 % of cases were men, the median age of the patients was (45.21±13.49) years and the annual incidence in our region was 67/100 000 per year. The diagnostic methods included pathogenic bacteria culture, serological diagnosis, and suspect case were 24.39 %, 47.56 %, and 28.05 %, respectively, sensitivity of combined detection Standard agglutination test (SAT) and the Rose Bengal test (RBT) is 96.2 %. In our work, 80 cases of brucellosis were diagnosed by a bacterial culture which were been identified as Brucella melitensis, blood culture was the main method (78.75 %) and the average positive alarm time was 80.74 (21.6–129) h and all of them were detected in aerobic bottles, followed by synovial fluid, bone marrow, lumbar spine, and joint tissue, puncture fluid and ascites culture which were 6.25 %, 3.75 %, 5.00 %, 5.00 % and 1.25 % respectively. The brucellosis with complications was lumbar spine lesions at 41.46 % cervical spine lesions at 4.60 % and knee joint lesions at 12.8 % and another osteoarthritis. The in-hospital mortality rate of the patients was 0.91 % and all of them were meningitis patients. ROC analysis indicated CRP had high sensitivity and specificity for brucellosis, and when CRP was 1.23mg/ml, the sensitivity and specificity were 0.727 and 0.718 respectively, and the U test also indicated CRP had a significant difference, Z=5.054, p <0.001. ConclusionsBrucellosis is frequently morbidity in 40 + age men, which has been diagnosed by aerobic blood culture, generally bacterial culture, RBT and SAT, epidemiological, and commonly with complications of spine and arthropathy.

Is there an Association between the ABO Blood type and Bleeding or Clotting Times?

Introduction: Evaluating blood type, bleeding time (BT), and clotting time (CT) are essential before surgery. Blood type reveals personality. Multiple studies link ABO blood types to digestive (GI) disorders, urinary tract infections (UTIs), cardiovascular diseases (CVD), diabetes, thrombosis, and epistaxis. BT and CT differ among blood types. This study investigated the relationship between blood types and BT and CT. Aims and Objectives: To investigate the connection between BT and CT and Blood Groups in medical students and identify gender differences in these parameters.Materials and Methods: This cross-sectional research was carried out between September 2021 and May 2022 on 250 healthy first-phase MBBS students (17–20 years old) of both sexes (batch 2020–2021 &amp; 2021-22) at GMC Jammu’s haematology laboratory. The blood group was determined using a standard antiserum Agglutination test—CT using Wright’s Capillary tube technique and BT using Duke’s filter paper technique. Based on the methodology above, we classify regular BT and CT as 2 to 6 minutes and 3 to 8 minutes, respectively. BT and CT levels were examined for each blood type. The blood group distribution was reported as a percentage, the relationship between BT or CT and blood groups was analysed using Chi-square analysis, and the gender differences between these parameters were analysed using an unpaired t-test in SPSS 26.0. Results: In our investigation, the distribution of ABO blood groups was determined to be: 39.6% B &gt; 34% O &gt; 21.2% A &gt; 5.2% AB. Chi-square analysis revealed a non-significant ABO blood group correlation with BT (p=0.938) and CT (p=0.652). Comparing blood types with BT and CT, the AB and B blood groups had higher mean BT and CT values among the study participants. The blood type influenced BT/CT. Screening and prevention must commence before the onset of bleeding disorders.&#x0D; Conclusion: The most prevalent blood group among medical students was B, whereas AB was the least prevalent. Blood types AB and B have greater levels of BT and CT. Neither BT nor CT were significantly associated with blood type. BT and CT values were found to be greater in females

Prevalence of AO blood group and level of agreement for AO blood-typing methods in pet pigs from Louisiana.

Pet pigs are a species of growing medical interest, and evidence-based practices for blood transfusions are needed. The objectives of this study were to quantify the prevalence of 3 blood group (Bg) phenotypes ("A" and "Aweak " resulting from EAAAA and EAAAO , "0" from EAA00 , or "-" from EAA00 or SSS alleles) in pet pigs and compare results using a human blood-typing card (EldonCard), standard saline agglutination (SSA), and polymerase chain reaction (PCR) sequencing. Cross-sectional study. University veterinary teaching hospital. Ninety-seven pet pigs from Louisiana. Blood was sampled from randomly selected pet pigs of various breeds, anticoagulated with EDTA, and typed using each investigated test according to the manufacturers' directions or standard laboratory technique. Samples for PCR analysis were stored at -80°C until analysis. Phenotypes were screened via EldonCard. Association between Bg and sex was investigated using chi-square test, with significance at P<0.05. Kappa (κ) statistic was used to measure the level of agreement between the 3 tests. Pot-bellied pigs represented the majority (84.5%) of this population, with 52 (53.6%) males and 45 (46.4%) females. Genotypic frequencies were 30%, 30%, and 40% for "EAAAA ," "EAAAO ," and "EAA00 ," respectively. Based on EldonCard, 38 phenotypes were classified as "A," 5 as "Aweak ," and 54 as "0" or "-." Results were identical for Bg, with the 3 tested techniques in 90% (45/50) of samples. Agreement between EldonCardand PCR was almost perfect (49/50 [98%], κ=0.959; P<0.001). Agreement between SSA and PCR, and EldonCardand SSA was substantial (46/50 [92%], κ=0.803, P<0.001 and 93/97 [95.9%], κ=0.764, P<0.001, respectively). The most common blood type was "0" or "-" (55.7%), followed by "A" (39.2%) and "Aweak " (5.1%). There was strong agreement between EldonCard and PCR testing. EldonCard allowed for rapid and reliable phenotype identification ("A," "Aweak ," and "0" or "-") and represents a clinically applicable laboratory method for blood typing in pet pigs.

Molecular and Serological Testing for Brucellosis in Egypt

Background: Brucellosis is highly endemic in the Mediterranean Basin, including Egypt. Despite attempts to control the disease in Egypt, there are still significant problems with diagnostic accuracy. The prevalence and incidence of brucellosis are unknown and we have uncertain predictive prognostic tests for brucellosis treatment results. Identification of the actual epidemiological burden of brucellosis in Egypt, and levels of the brucellosis antibody titer among rural and urban populations is critical to the evaluation of combined treatment approaches that achieve lesser relapse rates. Objective: To determine the current prevalence of brucellosis infections in Egypt and changes in its epidemiological pattern. Methods: We compared the available diagnostic yield, sensitivity, specificity, accuracy, cost, and time consumption of serological tests with those from quantitative polymerase chain reaction (qPCR) to establish their ability to meet the diagnostic criteria. Also, we conducted surveillance of the rates of brucellosis infection in both humans and animals. Results: The enzyme-linked immunosorbent assay (ELISA) and blood cultures were less sensitive diagnostic methods for the detection of brucellosis. These approaches are technically challenging and have a high likelihood of false negatives. Therefore, they are best reserved for suspected cases with negative standard agglutination test (SAT). Conclusion: A more practical approach to the diagnosis of brucellosis depends on epidemiological testing for risk factors, clinically suspected cases, and SAT titers ≥ 1/320. Strategies to prevent relapsing include: 1) Health education of patients, 2) Long-term triple therapy, e.g., three months, with possible extension to six months in severe or recurrent cases or when complications occur.

High-level seroprevalence against Leptospira interrogans serovars among wild foxes, jackals and stray dogs in the North Khorasan Province, Iran.

BackgroundLeptospirosis is an important, neglected zoonotic disease that affects people and animals in humid (sub)tropical regions. Wild canines carry the pathogen and may contaminate natural resources which may then act as a source of human infection.ObjectivesThe study was designed to understand the seroprevalence of leptospirosis among domestic and wild canines in Bojnurd County, Northeast Iran.MethodsA total of 77 serum samples, comprising 29 sera from asymptomatic wild canines [foxes (n = 25) and jackals (n = 4)] and 48 sera from asymptomatic stray dogs, was investigated. Serovars were identified and antibody titres were measured by standard microscopic agglutination test (MAT) using serial serum dilutions.ResultsAmong all serum samples, 44.1% reacted positively to a Leptospira interrogans serovars. The average percentage of positive reactions was higher in stray dogs than in wild canines although this did not reach statistical significance (55.2% and 37.5%, p = 0.159). Positive reactions with L. i. Pomona, L. i. Australis and L. i. Tarasovi was detected only among jackals and foxes. Among the stray dogs, the highest number of positive sera were for L. i. Grippotyphosa (61.1%) and L. i. Canicola (50%). The highest titre detected was for L. i. canicola (1:1600) in two stray dogs and against L. i. Icterohaemorrhagiae and L. i. Pomona (1:800) in a single jackal.ConclusionsThe study revealed that leptospirosis is endemic among various canine species in the North Khorasan Province of Iran. Detailed monitoring of canines is necessary for better understanding the epidemiology of infection in our and other Iranian regions.

Human brucellosis in the southern West Bank, 2015–17: a seroprevalence and molecular characterisation study

BackgroundBrucellosis is a global zoonotic disease ranked by WHO as being among the top seven neglected zoonoses. After implementing a livestock vaccination strategy in 1998, the incidence rate of human brucellosis in Palestine dropped significantly. However, in 2015 the disease re-emerged with alarming incidence rates similar to those in the period before 1998. The aim of this study was to characterise the strains associated with this episode using genetic and genomic tools. MethodsAll patients were referred to the department of preventive medicine from several public or private clinics in the southern West Bank due to undulant fever. After a careful analysis of a patients’ history to evaluate exposure to Brucella, blood samples from patients suspected of brucellosis were collected. Samples positive for the Rose Bengal test with an antibody titre of 1:80 IU/mL or more by Brucella standard agglutination test, and not under antibiotic treatment, were further cultured using the BACTEC 9240 blood culture system. DNA was extracted from the positive culture samples and analysed using three biomarkers to validate the presence of the Brucella genus, establish the species, and identify the Brucella melitensis Rev 1 vaccine strain. Nine randomly chosen isolates were sequenced using targeted next generation sequencing technology to screen for the rpoB gene mutations. Multiple-locus variable number of tandem repeats analysis (MLVA) with 16 variable number of tandem repeats markers was used to genotype the isolates and BioNumerics version 7.6 software was used to analyse the data using the categorical coefficient and unweighted pair group method using the arithmetic averages algorithm. FindingsBlood samples were obtained from a total of 1324 patients during 2015–17. 704 (53·2%) of 1324 serum samples showed agglutination in the Rose Bengal test and 559 (79·4%) of the 704 were considered seropositive based on a standard agglutination test titre of 1:80 IU/mL or more. Only 130 (23·3%) of the 559 seropositive samples were from patients not under antibiotic treatment. Upon culturing, only 72 (12·9%) of the seropositive samples showed growth signals and were positive by PCR for the Brucella genus marker, and all isolates were B melitensis. None of the samples were associated with the B melitensis Rev 1 vaccine strain. The results of sequencing the rpoB gene by next generation sequencing showed four different single nucleotide variations that were not located within the rifampicin resistance-determining region. Using MLVA-16 analysis and the MLVA bank benchmark, the studied isolates were divided into 22 unique genotypes. A hierarchical clustering analysis for the 72 isolates and 18 B melitensis global isolates representing four major regions in the Middle East, Europe, Africa, and America, clustered the 73 isolates in the Middle East clade, providing a similarity cutoff value of more than 97% for genotype and more than 75% for clade. InterpretationThis study shows that the brucellosis episode of 2015–17 was due to B melitensis local isolates and was not associated with the Rev 1 vaccine stain. The results show that MLVA-16 is a useful tool for the public health surveillance of human brucellosis. The present study analysed samples from the Hebron governorate only. It is recommended to include more human and animal samples from different areas in Palestine to understand the molecular epidemiology of brucellosis in Palestine and to make more effective decisions regarding control and treatment programmes. FundingPalestine Polytechnic University.

Prevalence of leptospirosis among patients attending renal and general outpatient clinics in Mulago Hospital, Kampala, Uganda

In this study, we sought to establish the prevalence of leptospirosis among renal patients and general outpatients attending Mulago National Referral Hospital, Uganda. A total of 254 patients were recruited, their blood samples collected and interviewer-administered semi-structured questionnaires provided between July and October 2018. These questionnaires captured data on sociodemographic characteristics and symptoms of leptospirosis disease. An individual with an average body temperature of 37.3 ± 1.1 °C was considered to be having fever. The blood samples were analyzed using the standard Microscopic Agglutination Test (MAT) with a panel of 14 Leptospira-serovars belonging to 11 serogroups. Prevalence was reported with confidence intervals while questionnaire data was analyzed using logistic regression analysis. We present an overall prevalence of leptospirosis at 4.70% (95% CI = 2.60–8.30) after analysis of samples from recruited patients. This seropositivity (12/254) was classified into 7 serovars, among which, Canicola and Djasiman presented with titers between ≥ 200 and ≥ 400 in samples of both renal patients and outpatients, indicative of the active disease. Djasiman was the highest contributor to the reported prevalence. Overall, most examined participants presented with common symptoms of abdominal pain (AOR = 24.4, 95% CI (2.42–267.89), p = 0.02) and dehydration (AOR = 0.1, 95% CI (0.01–0.69), p = 0.05). Our study suggests that these symptoms and previous history of abdominal pain may be caused by Leptospira infections among the studied participants. We therefore recommend inclusion of leptospirosis in the differential diagnosis for renal and febrile illnesses. Indeed, abdominal pain and dehydration should be further studied with a bigger sample size and for other related febrile illnesses.

HUMAN BRUCELLOSIS AMONG PYREXIA OF UNKNOWN ORIGIN CASES IN TWO TERTIARY HOSPITALS, IN SANA'A, YEMEN

Background and objectives: Brucellosis continues a most important health problem in numerous parts of the world and in the Middle East regions is a significant origin of acute febrile illness. This study was conducted to determine Brucella antibodies among patients with PUO, the association of brucellosis with demographic characteristics, identify clinical symptoms and risk factors for Brucella infection among PUO cases. Materials and Methods: This cross-sectional research study was conducted in two health institutions; Al-Jumhouri Hospital and Al-Thawra Hospital, between January 2021 to January 2022. The required data were gathered from each patient by using pre-defined standardized questionnaire. Five mL of sterile whole blood was collected from each patient and then tested for Brucella antibodies using a standard tubular agglutination test. The data were then analyzed using the statistical software Epi Info version 6. Results: Among the 241 PUO patients, females were 72.2% and males 27.8%. The mean ± SD for total age was 28.8±15.8 years. The rate of Brucella antibody among patients with PUO was 29%, females 78.6% higher than males by 21.4%. Clinical characteristics of PUO patients were persistent fever (36.9%), intermittent fever (63.1%), sweating (63.5%), shivering (61.4%), joint pain (78.4%), muscle pain (78.4%), back pain (74.7) back head pain (66.4%), headache (60.4%), weight loss (52.9%), body weakness (89.2%), loss of appetite (50.6%), and nausea (46.1%). The risk factors for brucellosis among the PUO patients was handling of animals during parturition (69.2%, OR=7, p&lt;0.0001). Conclusion: These findings revealed a high prevalence of human brucellosis among PUO patients in Sana'a city and will becoming a serious problem that threats the health care system in Yemen. So, awareness programs should be provided to doctors, population about Brucella infection and its risk factors. Peer Review History: Received: 4 January 2022; Revised: 16 February; Accepted: 10 March, Available online: 15 March 2022 Academic Editor: Dr. Amany Mohamed Alboghdadly, Princess Nourah bint abdulrahman university, Riyadh, amalbgadley@pnu.edu.sa UJPR follows the most transparent and toughest ‘Advanced OPEN peer review’ system. The identity of the authors and, reviewers will be known to each other. This transparent process will help to eradicate any possible malicious/purposeful interference by any person (publishing staff, reviewer, editor, author, etc) during peer review. As a result of this unique system, all reviewers will get their due recognition and respect, once their names are published in the papers. We expect that, by publishing peer review reports with published papers, will be helpful to many authors for drafting their article according to the specifications. Auhors will remove any error of their article and they will improve their article(s) according to the previous reports displayed with published article(s). The main purpose of it is ‘to improve the quality of a candidate manuscript’. Our reviewers check the ‘strength and weakness of a manuscript honestly’. There will increase in the perfection, and transparency. Received file: Reviewer's Comments: Average Peer review marks at initial stage: 6.0/10 Average Peer review marks at publication stage: 7.5/10 Reviewers: Dr. Bilge Ahsen KARA, Ankara Gazi Mustafa Kemal Hospital, Turkey, ahsndkyc@gmail.com Dr. Wadhah Hassan Ali Edrees, Hajja University, Yemen, edress2020@gmail.com Similar Articles: PATTERNS OF ANTIMICROBIAL RESISTANCE AMONG MAJOR BACTERIAL PATHOGENS ISOLATED FROM CLINICAL SAMPLES IN TWO TERTIARY’S HOSPITALS, IN SANA'A, YEMEN