Indirect Hemagglutination Assay Research Articles

Indirect hemagglutination assay for diagnosing brucellosis: Past, present, and future

Brucellosis is a zoonotic disease that causes enormous losses in livestock production worldwide and has a significant public health impact. None of the brucellosis-free countries is currently able to guarantee their ability to prevent the introduction of the pathogen due to the increase in tourism and the expansion of migration. The timely identification of infected animals is an effective means of preventing brucellosis and minimizing the epidemiological risk. The tube agglutination test, Rose Bengal plate test, complement fixation test, and enzyme-linked immunosorbent assay, which are routinely used to identify seropositive productive animals, have limitations and results that do not always correlate. The indirect hemagglutination assay (IHA) stands out among non-traditional methods because it is affordable, has a simple protocol, and is more reliable than classical serological tests, especially in cases of questionable and/or false-negative results. The diagnostic value of the IHA has long been studied by laboratories in several countries, but mostly by post-soviet research teams; therefore, the results continue to be published in Russian-language journals, ensuring that the local scientific community can access the results. In addition, the efficacy of this test for the diagnosis of brucellosis and other infectious diseases has not yet been reviewed. The purpose of this review was to summarize the results of studies on the development and use of IHA for the diagnosis of brucellosis and to determine the prospects for further improvement Keywords: brucellosis, diagnostic value, indirect hemagglutination assay, prospects for improving the test, serological tests.

Comparison of Methods in the Serologic Diagnosis of Cystic Echinococcosis.

Cystic echinococcosis (CE) is caused by the larval form of Echinococcus granulosus. Clinical, radiologic, pathologic, and serologic findings should be evaluated together for the diagnosis of CE. The sensitivity and specificity oalf serologic tests may vary depending on the method used. In this study, we aimed to detect IgG antibodies specific to E. granulosus using indirect hemagglutination assay (IHA), enzyme-linked immunosorbent assay (ELISA), indirect fluorescent antibodies (IFA) and western blot (WB) tests. In our study, the serum samples of 74 patients sent to our laboratory with suspicion of CE were studied using two different commercial IHA tests, ELISA, IFA and WB test. The test results were evaluated along with radiological findings and histopathological examinations, the latter being the gold standard. Of all the patients, 51 (69%) were female and 23 (31%) were male. There was a statistically significant difference between males and females (χ2 = 9.7, p = 0.002). Out of 74 patients, positivity rates for Siemens IHA, Fumouze IHA, ELISA, IFA and WB test were positive as 33 (44.6%), 35 (47.3%), 43 (58.1%), 42 (56.7%) and 38 (51.3%), respectively. The sensitivity and specificity of the tests were as follows: 66.67 and 2.31% for Siemens IHA; 70.83% and 96.15% for Fumouze IHA; 85.42%, and 88.46% for ELISA; 83.33% and 88.46% for IFA; 72.92% and 88.46% for WB test. There were statistically significant differences in between all five methods (p < 0,001). While the tests with the highest specificity was Fumouze IHA, the test with the highest sensitivity was the ELISA test. It was concluded that IHA and ELISA tests were more practical in practice because of their greater applicability.

USING INDIRECT HEMAGGLUTINATION ASSAY FOR THE DIAGNOSIS OF CATTLE BRUCELLOSIS

Abstract: The possibility of using indirect hemagglutination assay (IHA) with milk (IHA/m) to differentiate post-infectious antibodies from post-vaccination ones in cows immunized with the Brucella abortus 82 was studied. Anti-Brucella antibodies by IHA/m were detected in the milk of all lactating animals (n=20) culled on the results of tube agglutination test (AT) and/or complement fixation test (CFT), while the milk ring test (MRT) were negative in 20% of cases. IHA/m, as well as IHA with blood serum (IHA/s), confirmed brucellosis in cows with AT- and/or CFT- negative or questionable results. The correlation coefficient between the results of the two IHA variants was very high (r = 793), and the hemagglutinin titers in the blood serum were significantly higher - 1:760 (+13.3%; -11.7%) than in AT-1:260 (+8.7%;-8.0%) (P≤0.01). Vaccination of cows caused increased production of complement-fixing, agglutinating and precipitating antibodies, which tended to weaken during observation of the animals: 30, 60 and 90 days post-vaccination (p.v.). By the end of the experiment, all vaccinated animals showed questionable AT; positive RID/O-PS and IHA/s were noted in 10% and 20% of cases, respectively. However, complement-fixing antibodies remained at diagnostic values in all animals until the end of the experiment. Despite the intense antibody immune response to the vaccine injection, there were no Brucella-specific agglutinins detected by IHA/m and MRT in the milk of cows even on the 30th day p.v. The results show the need for further study on a large population to determine the diagnostic value of IHA/m for differentiating infected from vaccinated animals.&#x0D;

Capacity building in schistosomiasis control institutions in China: a cross-sectional study

To understand the current status of capacity building in schistosomiasis control institutes in schistosomiasis-endemic provinces (municipality, autonomous region) of China. The responsibilities and construction requirements of various schistosomiasis control institutions were surveyed by expert discussions, and field interviews and visits during the period between May and June, 2023, and the questionnaire for capacity maintenance and consolidation in schistosomiasis control institutions was designed. An online questionnaire survey was conducted in county-, municipal-, and provincial-level institutions that undertook schistosomiasis control and surveillance activities through the Wenjuanxing program. The distribution of schistosomiasis control institutions, the status of institutions, departments and staff undertaking schistosomiasis control activities and the translation of scientific researches on schistosomiasis control in China were analyzed. The laboratories accredited by China National Accreditation Service for Conformity Assessment (CNAS) were considered to be capable for testing associated with schistosomiasis control, and the testing capability of schistosomiasis control institutions was analyzed. A total of 486 valid questionnaires were recovered from 486 schistosomiasis control institutions in 12 endemic provinces (municipality, autonomous region) of China, including 12 provincial-level institutions (2.5%), 77 municipal-level institutions (15.8%) and 397 county-level institutions (81.7%). Of all schistosomiasis control institutions, 376 (77.4%) were centers for disease control and prevention or public health centers, 102 (21.0%) were institutions for schistosomiasis, endemic disease and parasitic disease control, and 8 (1.6%) were hospitals, healthcare centers or others. There were 37 713 active employees in the 486 schistosomiasis control institutions, including 5 675 employees related to schistosomiasis control, and the proportions of employees associated with schistosomiasis control among all active employees were 5.9% (231/3 897), 5.5% (566/10 134), and 20.6% (4 878/23 682) in provincial-, municipal-, and county-level institutions, respectively. There were 3 826 full-time employees working in schistosomiasis control activities, with 30.5% (1 166/3 826), 34.6% (1 324) and 34.9% (1 336/3 826) at ages of 40 years and below, 41 to 50 years and over 50 years, and there were 1 571 (41.0%) full-time schistosomiasis control employees with duration of schistosomiasis control activities for over 25 years, and 1 358 (35.5%) employees with junior professional titles and 1 290 with intermediate professional titles (35.5%), while 712 (18.6%) full-time employees working in schistosomiasis control activities had no professional titles. The three core schistosomiasis control activities included snail control (26.3%, 374/1 420), epidemics surveillance and management (25.4%, 361/1 420) and health education (18.8%, 267/1 420) in schistosomiasis control institutions. The Kato-Katz method, miracidium hatching test with nylon gauzes, and indirect haemagglutination assay (IHA) were the most commonly used techniques for detection of schistosomiasis, and there were less than 50% laboratories that had capabilities or experimental conditions for performing enzyme-linked immunosorbent assay (ELISA), dipstick dye immunoassay (DDIA), dot immunogold filtration assay (DIG-FA), loop-mediated isothermal amplification (LAMP) and polymerase chain reaction (PCR) assays. During the period from 2018 to 2022, schistosomiasis control institutions had undertaken a total of 211 research projects for schistosomiasis control, with a total funding of 18.596 million RMB, published 619 articles, participated in formulation of 13 schistosomiasis control-related criteria, and applied for 113 schistosomiasis control-related patents, including 101 that were granted, and commercialized 4 scientific research outcomes. The proportion of independent specialized schistosomiasis control institutions is low in schistosomiasis control institutions in China, which suffers from problems of unsatisfactory laboratory testing capabilities, aging of staff and a high proportion of low-level professional titles. More investment into and intensified schistosomiasis control activities and improved capability building and talent cultivation in schistosomiasis control institutions are recommended to provide a powerful support for high-quality elimination of schistosomiasis in China.

Seroprevalence of melioidosis and its association with blood profiles and pathogens in sheltered dogs in southern Thailand.

Melioidosis is a notable zoonotic disease in Thailand that can affect both humans and animals. Although dogs are one of the most popular pets worldwide, there is a remarkable lack of information on the prevalence and knowledge of canine melioidosis. This study aimed to estimate the seroprevalence of melioidosis in sheltered dogs and its relationship with the blood profile and blood pathogens. Melioidosis in 156 dogs was analyzed using an indirect hemagglutination assay. Hematology and serum biochemistry tests were performed using an automated system. Blood pathogens (e.g., Ehrlichia, Anaplasma, Hepatozoon, and Babesia) were diagnosed using conventional polymerase chain reaction. The seroprevalence rates of canine melioidosis and blood pathogen infection were 5.77% (9/156) and 50.64% (79/156), respectively. Seropositive dogs generally have higher lymphocyte counts and aspartate aminotransferase levels but lower total white blood cell, neutrophil, and platelet (PLT) counts than seronegative dogs. No statistically significant difference (p > 0.05) was observed between the seropositive and seronegative dogs' hematology and serum biochemistry findings. Neither the correlation between melioidosis and blood pathogen infection nor the association between melioidosis and thrombocytopenia was statistically significant (p > 0.05). Remarkably, dogs that had coinfections with both melioidosis and blood pathogens demonstrated a significantly reduced PLTcount (49,167 ± 7,167) compared with dogs that tested positive for melioidosis but negative for blood pathogens (139,333 ± 29,913) (p < 0.01). In southern Thailand, the prevalence of canine melioidosis was low but the prevalence of blood pathogens was high. Coinfection with blood pathogens can significantly reduce PLT counts, which may have a potentially serious impact. Future research should focus on conducting seroprevalence studies in the general dog population.

Seroprevalence and Risk Factors of Chlamydia Infection in Pigs in Hunan Province, Southern China, 2017-2018.

Background: Chlamydia is a Gram-negative obligate intracellular bacterium that is pathogenic for humans and a large variety of veterinary animal species. However, there is no continuous monitoring of chlamydia infection data in pigs in Hunan province, southern China. Therefore, in order to evaluate the seroprevalence and identify risk factors associated with Chlamydia infection in pigs within this region, a comprehensive study was conducted. Methods: A total of 3848 serum samples were collected from pigs (from farmers and companies) between May 2017 and August 2018. The presence of specific antibodies against Chlamydia was determined through the employment of the indirect hemagglutination assay (IHA). Results: The overall seroprevalence of Chlamydia was determined to be 26.90% (1038/3848, 95% confidence interval: 25.60-28.40). By employing statistical analysis using SPSS software (p < 0.05), factors such as altitude, sampling regions, and rearing systems of pigs were identified as potential risk factors for Chlamydia infection. Conclusion: These findings elucidate a substantial prevalence of Chlamydia in pigs within the mountainous region of Hunan province, southern China, thereby highlighting a potential risk to human health. These results underscore the need for proactive measures and targeted interventions to mitigate the transmission of Chlamydia in porcine populations, safeguarding both animal welfare and public health.

Clinical information associated with bacterial culture proven Burkholderia pseudomallei among melioiodosis patients in Buriram province

Indirect hemagglutination assay (IHA) is a general rapid diagnosis procedure in melioidosis patients combined with clinical information. The Burkholderia pseudomallei culture technique requires prolonged time before getting culture results. Related factors of bacterial culture proven melioidosis and the appropriate cut-off point for IHA are unfashionable and need to be updated. A cross-sectional study was conducted among melioidosis patients using data reported in the medical records. Univariable analysis was performed by Chi-square test and Student T-test as appropriate. Multivariable logistic regression was finally used to identify the contributing factors to bacterial culture proven cases. The results were shown as adjusted odds ratio (AdjOR) and 95%CI. Validity domains, sensitivity, specificity, predictive values, and likelihood ratios at each cut-off point for IHA were demonstrated. The results revealed that after adjustment for abdominal pain symptoms, diabetes mellitus (DM)(p-value &lt; 0.001) and dyspnea symptoms (p-value = 0.025) were the associated factors of bacterial culture proven cases. The IHA titer &gt; 1: 10240 provided the highest positive and negative predictive values, 30% and 69% respectively. Both positive and negative likelihood ratios of this cut-off point were also accounted for as 0.95 and 1.003, respectively. Health personnel should perform melioidosis diagnosis by the use of clinical information and an appropriate cut-off point for IHA, especially in diabetic patients who have dyspnea.

ELISA F29 -A therapeutic efficacy biomarker in Chagas disease: Evaluation in pediatric patients treated with nifurtimox and followed for 4 years post-treatment.

Measurement of the success of antitrypanosomal treatment for Chagas disease is difficult, particularly in the chronic phase of the disease, because anti-Trypanosoma cruzi antibodies persist in serum for prolonged periods. We studied the effects of nifurtimox administered by two different treatment regimens on the T. cruzi calcium-binding flagellar protein F29 in children diagnosed with Chagas disease measured using an enzyme-linked immunosorbent assay (ELISA) technique (ELISA F29). In a phase 3, randomized, double-blind, parallel-group, historically controlled study (ClinicalTrials.gov NCT02625974), blood samples obtained from children diagnosed with Chagas disease and treated with nifurtimox for either 60 days or 30 days were analyzed using an ELISA with an F29 recombinant protein as the antigen, as well as conventional serological tests (recombinant ELISA and indirect hemagglutination assay). In an exploratory approach, serological response to nifurtimox treatment was evaluated for 4 years post-treatment. In both treatment groups, the number of patients with negative ELISA F29 values increased over the period of observation. The incidence rate of negative seroconversion using ELISA F29 was 22.94% (95% CI: 19.65%, 26.63%) in the 60-day treatment group and 21.64% (95% CI: 17.21%, 26.86%) in the 30-day treatment group. In the subpopulation of patients who tested seropositive for F29 before nifurtimox treatment, 88 patients (67.7%) in the 60-day regimen and 39 patients (59.1%) in the 30-day regimen were F29 seronegative at 4 years post-treatment. All patients who had a positive ELISA F29 test at baseline and seroconverted to negative measured by conventional serology reached seronegativity in ELISA F29 earlier than in conventional serology. The results demonstrate a serological response to treatment with nifurtimox measured by the ELISA F29 test in children diagnosed with Chagas disease. The F29-based ELISA can be considered a potential early marker of response to antitrypanosomal therapy for Chagas disease. ClinicalTrials.gov NCT02625974.

Comparison of Melioidosis Indirect Hemagglutination Assay between Three Testing Laboratories in Australia.

Melioidosis caused by Burkholderia pseudomallei causes significant morbidity and mortality in Southeast Asia and northern Australia. Clinical manifestations remain diverse, including localized skin infection, pneumonia, and chronic abscess formation. Culture remains the gold standard of diagnosis, with serology and antigen detection tests playing a role if culture is unfeasible. Serologic diagnosis remains challenging, with limited standardization across different assays. In areas of endemicity, high rates of seropositivity have been documented. The indirect hemagglutination assay (IHA) is one of the more widely used serologic tests in these areas. In Australia, only three centers perform the test. Annually, laboratory A, laboratory B, and laboratory C perform approximately 1,000, 4,500, and 500 tests, respectively. A comparison of a total of 132 sera was analyzed from the routine quality exchange program between these centers from 2010 until 2019. Overall, 18.9% of sera tested had an interpretative discrepancy between laboratories. IMPORTANCE This study found significant discrepant results between three Australian centers offering the melioidosis indirect hemagglutination assay (IHA), despite testing the same samples. We have highlighted that the IHA is a nonstandardized test, which had different source antigens at each of the different laboratories. Melioidosis is a global disease, is associated with significant mortality, and is perhaps under recognized. It is likely to have increasing impact with changing weather patterns. The IHA has been used frequently as an adjunct to the diagnosis of clinical disease and is the mainstay of determining seroprevalence within populations. Despite its relative ease of use, especially in low resource settings, our study highlights the significant limitations of the melioidosis IHA. It has wide ranging implications, serving as an impetus for developing better diagnostic tests. This study is of interest to practitioners and researchers working in the various geographic regions affected by melioidosis.

Sentinel Surveillance of Schistosomiasis - China, 2021.

This report analyzes the national surveillance data for schistosomiasis in 2021 to understand the current status and provide evidence for further policy actions to promote elimination. This analysis is in line with the National Surveillance Plan of Schistosomiasis, which was revised in 2020 to adapt to the new stage of moving towards elimination. Data from the 2021 national surveillance of schistosomiasis in humans, livestock, and snails were collected from 13 provincial-level administrative divisions (PLADs) and analyzed using descriptive epidemiological methodology. The antibody-positive rate and area of newly discovered and re-emergent snail habitats were calculated. In 2021, a total of 31,661 local residents and 101,558 transient population were screened for antibodies using indirect hemagglutination assay (IHA). Of those who tested positive, 745 local residents and 438 transient population underwent further parasitological examination, with only one stool-positive result in the transient population. Additionally, 12,966 livestock were examined using the miracidia hatching test, with no positives detected. The total area of newly discovered and re-emergent snail habitats was 957,702 m2 and 4,381,617 m2, respectively. No infected snails were found using the microscopic dissection method, but six pooled snail samples were reported as positive using the loop-mediated isothermal amplification method for detecting specific sequences of Schistosoma. japonicum, in Anhui and Jiangxi Provinces. The prevalence of schistosomiasis among humans and livestock was found to be low, however, a potential transmission risk was identified in certain areas. To reduce the risk of transmission, a comprehensive control strategy should be continued and new techniques should be implemented in the surveillance and early warning system.

Is There an Association between Concurrent Epstein–Barr Virus Infection and Sudden Hearing Loss?—A Case—Control Study in an East Asian Population

Viral infection serves as the crucial etiology for the development of sudden sensorineural hearing loss (SSNHL). We aimed to investigate whether there is an association between concurrent Epstein-Barr virus (EBV) infection and SSNHL in an East Asian population. Patients who were older than 18 years of age and met the criteria of sudden hearing loss without an identifiable etiology were enrolled from July 2021 until June 2022, followed by the serological testing of IgA antibody responses against EBV-specific early antigen (EA) and viral capsid antigen (VCA) with an indirect hemagglutination assay (IHA) and real-time quantitative polymerase chain reaction (qPCR) of EBV DNA in serum before the treatment was initiated. After the treatment for SSNHL, post-treatment audiometry was performed to record the treatment response and degree of recovery. Among the 29 patients included during enrollment, 3 (10.3%) had a positive qPCR result for EBV. In addition, a trend of poor recovery of hearing thresholds was noted for those patients with a higher viral PCR titer. This is the first study to use real-time PCR to detect possible concurrent EBV infection in SSNHL. Our study demonstrated that approximately one-tenth of the enrolled SSNHL patients had evidence of concurrent EBV infection, as reflected by the positive qPCR test results, and a negative trend between hearing gain and the viral DNA PCR level was found within the affected cohort after steroid therapy. These findings indicate a possible role for EBV infection in East Asian patients with SSNHL. Further larger-scale research is needed to better understand the potential role and underlying mechanism of viral infection in the etiology of SSNHL.

Accuracy of Three Serological Techniques for the Diagnosis of Imported Schistosomiasis in Real Clinical Practice: Not All in the Same Boat

Schistosomiasis is a neglected tropical disease despite of being a major public health problem affecting nearly 240 million people in the world. Due to the migratory flow from endemic countries to Western countries, an increasing number of cases is being diagnosed in non-endemic areas, generally in migrants or people visiting these areas. Serology is the recommended method for screening and diagnosis of schistosomiasis in migrants from endemic regions. However, serological techniques have a highly variable sensitivity. The aim of this study was to evaluate retrospectively the sensitivity of three different serological tests used in real clinical practice for the screening and diagnosis of imported schistosomiasis in sub-Saharan migrant patients, using the detection of schistosome eggs in urine, faeces or tissues as the gold standard. We evaluated three different serological techniques in 405 sub-Saharan patients with confirmed schistosomiasis treated between 2004 and 2022: an enzyme-linked immunosorbent assay (ELISA), an indirect haemagglutination assay (IHA) and an immunochromatographic test (ICT). The overall sensitivity values obtained with the different techniques were: 44.4% for IHA, 71.2% for ELISA and 94.7% for ICT, respectively. According to species, ICT showed the highest sensitivity (S. haematobium: 94%, S. mansoni: 93.3%; and S. intercalatum/guineensis: 100%). In conclusion, our study shows that Schistosoma ICT has the best performance in real clinical practice, when compared to ELISA and IHA, in both S. mansoni and S. haematobium infections.

Seroprevalence and Risk Factors of Chlamydia Infection in Cattle in Shanxi Province, North China.

The information on Chlamydia infection in cattle is limited in Shanxi Province, north China. This study aimed to investigate the seroprevalence and risk factors of Chlamydia and Chlamydia abortus infection in cattle in Shanxi Province. In November 2020, a large-scale investigation of Chlamydia seroprevalence was conducted on 981 cattle serum samples collected from 40 cattle farms in 11 cities of Shanxi Province. The seroprevalence of Chlamydia and C. abortus was examined by indirect hemagglutination assay (IHA) and enzyme-linked immunosorbent assay (ELISA), respectively. The seroprevalence of Chlamydia and C. abortus was 52.29% (513/981) and 2.96% (29/981), respectively, in cattle in Shanxi Province. Location was identified as a risk factor for Chlamydia and C. abortus infection (p &lt; 0.05). Under different management patterns, the seroprevalence of Chlamydia and C. abortus in large-scale animal farming companies was higher than that in household animal farms and animal farming cooperatives, and only the seroprevalence of Chlamydia was significantly different in different management patterns (p &lt; 0.01). The results showed that there was higher seroprevalence of Chlamydia in cattle in Shanxi Province, while C. abortus was not the dominant species. This study provided baseline information on Chlamydia infection in cattle in Shanxi Province, which constitutes valuable data for monitoring livestock health and preventing potential zoonoses.

SERO-EPIDEMIOLOGY OF MEASLES IN CHILDREN FROM DISTRICT FAISALABAD PAKISTAN

A comparative cross sectional study was conducted on blood samples (n=200)collected from children of 1 to 10 years of age, selected by convenient sampling method. Differentcoupling agents were used to bind measles antigen with erythrocyte suspensions of differentconcentrations collected from different species for standardization of Indirect Hemagglutination Assay(IHA) which was then used for testing of serum samples for the detection and quantification antimeasles antibodies. Overall prevalence of anti-measles antibodies was 93.5 percent in target population. A statistically significant association was found between vaccinated and non-vaccinated individuals with appreciably high antibody titres in vaccinated individuals (P &lt; 0.01). Measles vaccine coverage was approximately 60 percent nationwide a complete mismatch to the global standards, andwas lowest among all other vaccines included in the Expanded Program on Immunization (EPI).Moreover, IHA proved to be a simple and convenient tool for serodiagnosis and for monitoring ofprotective humoral immune response against measles vaccine.

Clinical Manifestations and Etiology of Liver Abscess in Children in Tertiary Care Hospital

Background: The liver is a major metabolic organ with a dual blood supply, hence vulnerable to infection. The liver abscess (LA) is a space-occupying lesion in the liver parenchyma filled with pus caused by bacteria, parasites, or fungus. Objective: To determine the clinical manifestations and etiology of liver abscess among children presenting at a tertiary care hospital. Methodology: This descriptive cross-sectional study was conducted at the medical department of the National Institute of Child Health, Karachi, Pakistan from July 2022 to December 2022. A total of 147 children of either gender aged below 12 years with liver abscesses were analyzed. At the time of enrollment, demographic features, clinical features, characteristics, and etiological agents of LA. The cause of the liver abscess was confirmed by pus culture, blood culture, and indirect haemagglutination assay (IHA). Aspiration was performed among cases who had LA&gt;5cm and liquefication of abscess. Results: Of a total of 147 children with LA, 96 (65.3%) were boys. The mean age was 7.08±2.61 years. Socioeconomic status of 112 (76.2%) children was poor. In terms of frequency of presenting features/complaints, pyrexia (100%) and abdominal pain (98.6%) were the most commonly found whereas hepatomegaly (98%) and right hypochondrium tenderness (97.3%) were the commonest signs. The right hepatic lobe was involved in 120 (81.6%) LA cases and the majority of LA were solitary (93.2%). The size of the liver in 57 (38.8%) LA cases was between 3-6 cm while 93 (63.3%) cases were aspirated. The etiological diagnosis revealed that bacterial involvement was found in 36 (24.5%) LA cases while amoebic presence was noted in 75 (51.0%) cases. No cases of fungal involvement were found. Conclusion: A predominance of male gender and low socioeconomic status was found among children with LA presenting at a tertiary care hospital in Karachi, Pakistan. The most frequent presenting complaints were pyrexia and abdominal pain. Amoebic presence was the most common etiologic agent behind LA.

The Seroprevalence of Chlamydia Infection in Sheep in Shanxi Province, China.

Chlamydia, an obligate intracellular bacterium, can cause chlamydiosis in humans and animals worldwide and also leads to serious economic losses to the sheep industry. However, the information on Chlamydia infection in sheep was limited in Shanxi Province, northern China. In the present study, a total of 984 serum samples of sheep were collected from 11 regions in Shanxi Province, northern China in the autumn of 2020. The antibodies against Chlamydia and Chlamydia abortus were examined by the indirect hemagglutination assay (IHA) and indirect enzyme-linked immunosorbent assay (ELISA), respectively. The result showed that 351 (35.67%, 95% CI 32.68-38.66) of 984 serum samples were positive for Chlamydia, and the seroprevalence ranged from 6.67% to 70.79% among the different regions. In addition, antibodies to C. abortus infection were detected in 78 (7.93%, 95% CI 6.24-9.61) of 984 serum samples, and the seroprevalence ranged from 6.24% to 14.81% among the different regions. This is the first report on the seroprevalence of Chlamydia and C. abortus in sheep in Shanxi province, northern China. The findings provide baseline information for preventing and controlling Chlamydia infection in sheep in Shanxi Province, China.

Prevalence of Toxoplasmosis in Sheep and Goats in Pakistan: A Systematic Review and Meta-Analysis.

Toxoplasmosis, a parasitic disease caused by Toxoplasma gondii, results in congenital disorders and miscarriages among livestock and humans worldwide. This systematic review and meta-analysis were conducted to determine the prevalence of T. gondii infection in sheep and goats in Pakistan from 2000 to 2020. We searched the PubMed, Scopus, EMBASE, and Google Scholar databases and selected 17 publications that fulfilled our inclusion criteria. Eight studies were conducted in Southern Punjab, six in Khyber Pakhtunkhwa, two in Northern Punjab, and one in Central Punjab. The diagnostic tests used in the included articles to confirm toxoplasmosis were the latex agglutination test in 56% of the studies, the enzyme-linked immunosorbent assay in 38%, and the indirect hemagglutination assay in 6%. The infection rates were substantially higher among sheep &gt; 1 year of age (37%) than among sheep ≤ 1 year old (19%). Statistically significant differences in infection rates were found between male and female sheep and goats. The overall infection rate by age was also significant among sheep and goats. Sex and age variability between sheep investigations were significant, and sex heterogeneity and age homogeneity were significant among goats. Hence, robust infection control protocols should be implemented to prevent infection in animals and humans.

The Influence of Landscape Structure on the Occurrence of Neospora caninum, Toxoplasma gondii, and Sarcocystis spp. in Free-Living Neotropical Primates.

Habitat fragmentation is the main threat to primate survival in the world. Additionally, changes in the environments in which they live can also contribute to exposure to pathogens. To investigate some pathogens that free-living primates may be exposed to in Rio Grande do Sul State (RS; southern Brazil) and characterize the forest remnants in which they live, we investigated anti-Neospora caninum, Toxoplasma gondii, and Sarcocystis spp. antibodies in the serum of the animals. We analyzed 105 serum samples from 63 black howler monkeys (Alouatta caraya), 39 southern brown howler monkeys (Alouatta guariba clamitans), and 03 capuchin monkeys (Sapajus nigritus cucullatus), which were captured in forest fragments of RS. Indirect fluorescence antibody test (IFAT) and indirect hemagglutination assay (IHA) were used to detect antibodies to the agents. We then characterized the landscapes in a multiscale approach in radii from 200 to 1400m to investigate the relationship of the presence of the agents with landscape elements. In the IFAT-IgG, 13.3% (14/105) of the samples were seropositive for N. caninum, 4.8% (5/105) for T. gondii, and 5.7% (6/105) for Sarcocystis spp. In the IHA-IgM/IgG, 24.8% (26/105) were seropositive for T. gondii. The metrics that best explained exposure to agents were edge and patch density, forest cover, urban cover, and average Euclidean distance to the nearest patch. This study indicated that the primates were exposed to the agents studied, demonstrating that some landscape features are associated with exposures to the investigated pathogens.

Antimicrobial, antioxidant, antiviral activity, and gas chromatographic analysis of Varanus griseus oil extracts.

There is an urgent need to develop natural antimicrobials for the control of rapidly mutating drug-resistant bacteria and poultry viruses. Five extracts were prepared using diethyl ether, ethyl acetate, methanol, 1-butanol and n-hexane from abdominal fats of Varanus griseus locally known as Indian desert monitor. Antibacterial, antioxidant and antiviral activities from oil extracts were done through disc diffusion method, stable 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging assay and in ovo antiviral assay, respectively. The gas chromatography mass spectrometry (GC-MS) analyses were used to determine principal active compounds and chemical profile of each oil extract. n-Hexane extract showed clear zones of inhibition (ZOI) against Staphylococcus aureus, Escherichia coli and Klebsiella pneumoniae (12 ± 0.5mm, 9 ± 0.5mm, and 9 ± 0.5mm) while diethyl ether extract exhibited significant antibacterial activity (11 ± 0.5mm) against Proteus vulgaris only. In case of drug-resistant strains, methanol extract was active (6 ± 0.5mm) against Staphylococcus aureus, whereas n-hexane extract has shown ZOI 11 ± 0.5mm against P. aeruginosa. Range of percentage scavenging activity of V. griseus oil extracts from DPPH free radical assay was 34.9-70.7%. For antiviral potential, growth of new castle disease virus (NDV) was effectively inhibited by all five extracts (HA titer = 0-4). The highest antiviral activity against avian influenza virus (H9N2) was observed from methanol, diethyl ether and 1-Butanol oil extracts with HA titers of 2, 2 and 0, respectively. Methanol, diethyl ether, 1-butanol and n-hexane oil extracts produced best hemagglutination assay (HA) titer values (0, 0, 4 and 0) against infectious bronchitis virus (IBV). Ethyl acetate and 1-Butanol extract exhibited good antiviral potential against infectious bursal disease virus (IBDV) with indirect hemagglutination assay (IHA) titers of 8 and 4, respectively. Main classes of identified compounds through gas chromatography were aldehydes, fatty acids, phenols and esters. GC-MS identified 11 bioactive compounds in V. griseus oil extracts. It is summarized that V. griseus oil has strong antioxidant activity and good antimicrobial potential because of its bioactive compounds.

Comparison of a commercial ELISA and indirect hemagglutination assay with the modified agglutination test for detection of Toxoplasma gondii antibodies in giant panda (Ailuropoda melanoleuca).

Toxoplasma gondii is a worldwide-distributed zoonotic protozoan parasite which causes toxoplasmosis and has a significant effect on public health. In the giant panda (Ailuropoda melanoleuca), toxoplasmosis can cause asymptomatic infections, reproductive disorder and even death, which poses a serious threat to the conservation of this rare protected species. Therefore, serological investigation of T. gondii is essential to understanding its risk to giant pandas, however, there are no specific testing kits for giant pandas. Previous research has used MAT as the reference method for screening T. gondii, to investigate this further, this study focused on the agreement comparing of MAT with ELISA and IHA tests for detecting T. gondii antibodies in 100 blood samples from 55 captive giant pandas in Chengdu, China. The results showed 87.0%, 87.0%, 84.0%, samples were sero-positive for T. gondii using ELISA (kits a, b, c), respectively, while MAT and IHA tests were 84.0% and 9.0% sero-positive, respectively. There was no significant difference between MAT and the three ELISA kits and these two methods had substantial agreement (0.61 < қ ≤ 0.80). Meanwhile, there was a significant difference (P < 0.001) between MAT and IHA, and these two methods had only a slight agreement (қ ≤ 0.20). The relative sensitivity of the ELISA (kits a, b, c) were 89.0%, 91.5% and 95.1%, and the specificity were 86.7%, 80.0% and 80.0%, respectively, which showed these three ELISA kits all had great accuracy. It is suggested that MAT is the recommended test method for primary screening T. gondii in giant pandas and then verified by ELISA.