Serological and molecular detection of Toxoplasma gondii and Neospora caninum in free-ranging rats from Nagpur, India.

Toxoplasma gondii and Neospora caninum are cyst-forming coccidian parasites that infect both wild and domestic non-felids as intermediate hosts, with rodents serving as important reservoir hosts during their life cycles. This study was aimed at investigating T. gondii and N. caninum infections and identifying factors favouring T. gondii infection in free-ranging rats from India. A total of 181 rodents were trap-captured, and blood and brain samples were subsequently collected for serological and molecular examination of T. gondii and N. caninum. Antibodies against T. gondii and N. caninum were detected by MAT/NAT and IFAT in 13.8% (25/181) and 1.65% (3/181) of rodents, respectively. All three N. caninum samples positive by NAT/IFAT were also positive for ELISA, while for T. gondii, 19 of 25 MAT/IFAT positive samples were also positive for ELISA. The antibody titers (MAT/NAT/IFAT) of rodents seropositive for T. gondii ranged from 25 to 400, while those of rats seropositive for N. caninum ranged from 25 to 100. Also, using PCR, DNA from T. gondii (B1 gene) and N. caninum (NC5 gene) was found in 2.76% (5/181) of brain samples and 0.55% (1/181) of brain samples. All PCR positive samples were also seropositive. No mixed infections were observed in the serological and molecular detections. A Chi-square analysis revealed that older rats and rats living in urban areas are significantly associated with T. gondii infection; however, rodent species, gender, location, habitat types, and seasonality were statistically nonsignificant. Overall, this study demonstrated that T. gondii was widely distributed while N. caninum was less prevalent among free-ranging rats in the studied area.

Potential of cinnamaldehyde essential oil as a possible antimicrobial against fowl typhoid in layers.

Fowl typhoid (FT) is an economically significant bacterial disease of layers leading to a drastic drop in egg production. Due to increased public health concerns about antibiotics in poultry feed, a search for new safe antimicrobials for treating fowl typhoid is crucial. The antimicrobial effect of cinnamaldehyde essential oil (CnEO) against fowl typhoid in layers was investigated in this experiment. The 60-week-old BV300-layer birds (n = 100) were divided into five groups: the non-challenged control group A, only cinnamaldehyde-treated group B (CnEO @ 1:8000 dilutions through drinking water for 60 days), the challenged group C, challenged plus cinnamaldehyde therapy group D (CnEO @ 1:8000 dilutions through drinking water from 16 to 30 dpi), and challenged plus antibiotic therapy group E (chloramphenicol @ 1 gm/5lit through drinking water from 16 to 30 dpi). Hens from all challenged groups were challenged with Salmonella Gallinarum (VTCCBAA588) @ 1 × 108 CFU/ml orally. Various parameters such as clinical signs, mortality, egg production and egg weight, colony-forming unit (CFU) count of cecal content, eggshell surface, and egg yolk were evaluated all through 60 days of an experimental trial. Results indicated that, in the case of the cinnamaldehyde therapeutic group, there was a significant improvement in egg production, mild clinical signs, lower feed conversion ratio (FCR), and a significantly lower bacterial count in ceca and on the eggshell surface compared to the control challenge group. Thus, CnEO @ 1:8000 dilutions through drinking water can be a potential antimicrobial for controlling fowl typhoid.

Seroprevalence, risk factors, and serological cross-reactivity for diagnosis of Toxoplasma gondii and Neospora caninum infections in goats in India.

Toxoplasma gondii and Neospora caninum are genetically related cyst-forming protozoan parasites that cause reproductive failures in ruminants. Given the limited information on the epidemiology of these infections in goats in India, the study aimed to estimate the seroprevalence, assess antibody cross-reactivity for diagnosis, and identify associated risk factors. A total of 695 sera were evaluated for antibodies to T. gondii and N. caninum infections using Modified Agglutination Test (MAT for Toxoplasma)/Neospora agglutination test (NAT), Enzyme-linked immunosorbent assay (ELISA), and Indirect Fluorescent Antibody Test (IFAT for tachyzoite and bradyzoite stages). The seroprevalence rate of T. gondii and N. caninum infections was 56.9% and 10.9%, respectively. Inter-rater agreement (kappa value - κ) was calculated to assess agreements between various diagnostic assays, using the IFAT as the gold standard (for detecting both infections), the agreements for MAT/NAT (κ = 0.97) and the ELISA (κ = 0.95) were excellent. The acute infection among seropositive goats were determined using serological (IgG avidity test - measures the binding strength between IgG antibodies and parasite antigens) and molecular diagnoses (PCR for repetitive DNA sequences - Toxoplasma B1 gene: 131 bp and Neospora NC5 gene: 328 bp). Among seropositive goats ≥80% had high IgG avidity and <10% of animals had low IgG avidity antibodies for both infections. Most low IgG avidity goats were PCR positive for the TgB1 gene (94.4%) or Nc5 gene (85.7%). In the serological assays, we used different dilutions of test serum to rule out the cross-reactivity owing to similar antigenic makeup between these two parasites. When the serological cross-reactivity was analyzed using invasion assay at a serum titer of ≥200, more than 90% T. gondii positive sera showed host cell invasion of N. caninum and vice versa. Largely, the serological results indicate that cut-off serum dilution of ≥1:200 for ELISA and IFAT and ≥1:25 for MAT/NAT avoids serological cross-reactivity between T. gondii and N. caninum. Further, the Univariate and multivariate analyses showed that adult animals (>2 years), reservoir hosts, and extensive rearing systems are common risk factors for these infections. However, the history of abortion was identified as a significant risk factor for T. gondii infection. This study revealed that T. gondii and N. caninum infections are highly prevalent in this region and the use of an appropriate cut-off serum dilution is necessary to avoid cross-reactivity between these closely related parasites.

Toxoplasma gondii induces robust humoral immune response against cyst wall antigens in chronically infected animals and humans.

Toxoplasma gondii differentiation from proliferating tachyzoites into latent bradyzoites is central to pathogenesis and transmission. Strong humoral immune response has been reported against tachyzoite antigens, however, antibody-mediated response towards bradyzoite antigens is poorly characterized. This work aimed to study the humoral immune response towards bradyzoite and associated cyst wall antigens particularly CST1. The immunoreactivity of 404 goats, 88 sheep and 92 human sera to recombinant (CST1 and SRS9) and native proteins of encysted bradyzoite along with well-established tachyzoite antigens (SAG1 and GRA7) was determined using ELISA, Western blot and immunofluorescence analysis (IFA). ELISA results revealed nearly 50% of sera contain T. gondii specific antibodies. Results were further validated using Western blot and IFA. T. gondii positive sera predominantly recognized the cyst wall besides the known tachyzoite surface antigens. The presence of CST1 antibodies in seropositive samples were in line with the staining patterns which were consistent with CST localization. Notably, T. gondii IgM- IgG+ sera recognize the cyst wall whereas IgM + IgG-sera recognize tachyzoite antigens indicating acute infection consistent with presence of parasite DNA. The study demonstrates a strong humoral response against bradyzoite associated cyst wall antigens across naturally infected animals and humans. CST1 emerged as a key immunomodulatory antigen which may have direct implications for clinical immunodiagnostics.

Journey towards National Institute of One Health in India.

BACKGROUND & OBJECTIVES: Issues such as emerging and re-emerging infectious diseases, antimicrobial resistance, food security, biosafety and biosecurity are associated with changes in land use, population growth, urbanization, global travel and trade and climate change. As a result, a trans-disciplinary approach among human, animal and environmental health disciplines gained support. The Indian Council of Medical Research (ICMR) and Indian Council of Agricultural Research (ICAR) decided to establish a National Institute of One Health at Nagpur, Maharashtra, India. In this context, two collaborative research projects, funded by the ICAR and ICMR were initiated to conduct the epidemiological surveillance of selected zoonotic diseases in Central India. METHODS: Disease surveillance and molecular detection employing standard techniques like enzyme linked immunosorbent assay (ELISA), immuno-fluroscent assay (IFA), standard tube agglutination test (STAT) , Rose Bengal plate test (RBPT) and polymerase chain reaction (PCR) were undertaken based on the disease to be screened. RESULTS: In animals, the seropositivities for listeriosis (7.66%) and brucellosis (11.69%) were recorded. The occurrence of tuberculosis (3.8%) and leptospirosis (6.33%) was detected by PCR. Through cross-sectional studies from suspected human population with associated risk factors for zoonotic diseases, the seropositivity of brucellosis (1.83-11%), listeriosis (1.01-10.18 %), leptospirosis (8.14-12.67%) and scrub typhus (1.78-20.34%) was recorded. The investigations on scrub typhus indicated bimodal pattern during the months of pre-monsoon and post-monsoon season with a peak in post-monsoon in human cases. Ornithonyssus bacoti mites were identified from the rodents as a vector harbouring Orientia tsutsugamushi. The bovine tuberculosis was detected in 1.43 per cent human cases employing molecular assay. INTERPRETATION & CONCLUSIONS: The data indicated the occurrence of important zoonotic diseases adversely affecting the livestock health and human wellbeing. The scientific collaboration between veterinary and medical faculties has set an example for effective implementation of One Health (OH) programme for the establishment of National Institute of OH.

Cultivable microbiota and pulmonary lesions in polymicrobial bovine pneumonia.

In the present study, the spectrum of bacterial pathogens in the nasal shedding during disease process and in pneumonic lungs of dead animals was studied. A total of 288 clinical samples from cattle and buffaloes comprising of nasal swabs, blood, tracheal swabs, heart blood and lung tissue samples were collected from diseased (n = 190) and dead animals (n = 98). The recovered bacterial isolates were characterized by biochemical reactions, Matrix Assisted Laser Desorption/Ionization Time of Flight Mass Spectrometry (MALDI TOF-MS) and the 16S rRNA sequence analysis. The predominant bacterial isolates associated were Pasteurella multocida, Pseudomonas aeruginosa, Escherichia coli, Klebsiella pneumoniae and Staphylococcus aureus. The emerging pathogens causing bovine pneumonia identified were Leclercia spp., Stenotrophononas maltophila and Staphylococcus sciuri. Bacteriological examination of pneumonic lungs samples revealed 96.9% samples to be positive for polymicrobial isolation. Macroscopical lesions of lungs exhibited various stages and types of pneumonia with variable degree of haemorrhages, oedema and emphysema. Histopathologically, the fibrinous bronchopneumonia was observed to be the most frequent lesions seen in bovine pneumonia. Multi-drug resistance (MDR) was observed in 10% of P. multocida isolates. The resistance was seen for penicillin, cephalosporins and fluoroquinolones. Multi-drug resistance was seen in 90% of the E.coli tested. K. pneumoniae, E. hormaechei, E. cloacae, P. putida and Leclercia spp. identified were found to be multi-drug resistant. Understanding the etiological diversity of bacterial pathogens of bovine pneumonia may provide information for the better choice of therapeutics and health management.

Listeria goaensis sp. nov.

Two Listeria-like isolates obtained from mangrove swamps in Goa, India were characterized using polyphasic combinations of phenotypic, chemotaxonomic and whole-genome sequence (WGS)-based approaches. The isolates presented as short, non-spore-forming, Gram-positive rods, that were non-motile, oxidase-negative, catalase-positive and exhibited α-haemolysis on 5 % sheep- and horse-blood agar plates. The 16S rRNA gene sequences exhibited 93.7-99.7 % nucleotide identity to other Listeria species and had less than 92 % nucleotide identity to species of closely related genera, indicating that the isolates are de facto members of the genus Listeria. Their overall fatty acid composition resembled that of other Listeria species, with quantitative differences in iso C15 : 0, anteiso C15 : 0, iso C16 : 0, C16 : 0, iso C17 : 0 and anteiso C17 : 0 fatty acid profiles. Phylogeny based on 406 core coding DNA sequences grouped these two isolates in a monophyletic clade within the genus Listeria. WGS-based average nucleotide identity and in silico DNA-DNA hybridization values were lower than the recommended cut-off values of 95 and 70 %, respectively, to the other Listeria species, indicating that they are founding members of a novel Listeria species. We suggest the name Listeriagoaensis sp. nov. be created and the type strain is ILCC801T (=KCTC 33909;=DSM 29886;=MCC 3285).

Discontinuation of disease-modifying anti-rheumatic drugs and clinical outcomes in the Rheumatoid Arthritis DMARD Intervention and Utilisation Study 2 (RADIUS 2).

OBJECTIVES: The purpose of this analysis was to examine discontinuation and reasons for discontinuation from disease-modifying anti-rheumatic (DMARD) therapies in the RADIUS 2 registry, a long-term, open-label, observational study of patients with moderate to severe rheumatoid arthritis (RA). METHODS: Patients who participated in RADIUS 2 initiated etanercept (ETN) therapy at study entry and were followed for 5 years. In this post hoc analysis, patients who had received ETN continuously from entry to month 4 were categorised by treatment at month 4: ETN monotherapy, ETN+methotrexate (MTX), ETN+MTX+other DMARDs (OTH), or ETN+OTH. Outcomes were assessed at month 4 and at the time of any subsequent treatment change, and included Clinical Disease Activity Index (CDAI) and Health Assessment Questionnaire Disability Index (HAQ-DI). RESULTS: Of 3,484 patients analysed (982 ETN; 1,356 ETN+MTX; 537 ETN+MTX+OTH; 609 ETN+OTH), baseline demographic and clinical characteristics were similar across treatments. No treatment change occurred in 62.3%, 49.9%, 33.3%, and 37.1% of ETN, ETN+MTX, ETN+MTX+OTH, and ETN+OTH patients, respectively. The mean time on therapy from month 4 was longer for patients receiving ETN (23.3 months) or ETN+MTX (23.7 months) than those receiving ETN+MTX+OTH (18.0 months) or ETN+OTH (18.3 months). The greatest improvements in CDAI and HAQ-DI were seen in patients who continued on ETN. The most common reasons for discontinuing DMARD therapy were cost and ineffective treatment. CONCLUSIONS: Most patients who had received ≥4 months of ETN continued on ETN throughout the 5-year observation period. Patients with greatest clinical and disability improvements tended to continue on ETN.

Isolation and Seroprevalence of Aeromonas spp. Among Common Food Animals Slaughtered in Nagpur, Central India.

Aeromonads are ubiquitous foodborne pathogens with a global distribution. Animal-origin foods and contaminated animals are the main sources of Aeromonas infection to humans. So far little is known about the occurrence of Aeromonas spp. in food-producing animals in India. The present study was conducted to determine the prevalence and seroprevalence of Aeromonas species from 50 each of meat, blood, and sera samples collected from cattle, buffaloes, goats, and pigs slaughtered in and around Nagpur, Central India. Alkaline peptone water and ampicillin dextrin agar were used to isolate Aeromonas spp. An indirect enzyme-linked immunosorbent assay (ELISA) was standardized by use of whole-cell antigen (WC) and outer membrane protein (OMP) of Aeromonas hydrophila (MTCC 646). Aeromonads were isolated from 44 (22%) of the meat samples, and 1 (0.5%) from the blood samples. Seroprevalence by indirect ELISA-based WC antigen was estimated as 68% in cattle, 44% in buffaloes, 60% in goats, and 30% in pigs. OMP-based ELISA yielded a seroprevalence of 56%, 48%, 52%, and 22% in cattle, buffaloes, goats, and pigs, respectively. The results revealed that OMP-based ELISA and WC-based ELISA were in agreement with one another. Isolation along with high seropositivity demonstrates the presence of foodborne Aeromonas spp. in the Nagpur city of Central India.

Older age at rheumatoid arthritis onset and comorbidities correlate with less Health Assessment Questionnaire-Disability Index and Clinical Disease Activity Index response to etanercept in the RADIUS 2 registry.

BACKGROUND: Controversy exists in understanding the effects of age at onset and comorbidities in predicting rheumatoid arthritis (RA) response to biologic therapy. OBJECTIVE: The objective of this study was to investigate the influence of age at onset and number of comorbidities on Health Assessment Questionnaire-Disability Index (HAQ-DI) and Clinical Disease Activity Index (CDAI) responses in active RA patients after 6 months of treatment with etanercept. METHODS: One thousand eight hundred ninety-nine RA patients were assessed after 6 months of etanercept therapy. Patients met the following inclusion criteria: initiated etanercept, continued therapy for at least 6 months, and were not in CDAI low disease activity (LDA) at baseline (CDAI ≤10.0). Changes in HAQ-DI and CDAI scores over 6 months were analyzed across age of onset quintiles. Multivariate regression models evaluated the independent association between both age at onset and number of comorbidities with change in HAQ-DI/CDAI scores or achieving LDA, while accounting for other covariates. RESULTS: Significant improvements in HAQ-DI and CDAI scores were observed in all age-onset groups, although HAQ-DI improvements were less in older-onset patients. Results of multiple linear regression demonstrated that younger age at onset, higher baseline HAQ-DI/CDAI score, rheumatoid factor positivity, shorter disease duration, and fewer comorbidities at baseline were independently associated with improvement in both HAQ-DI and CDAI scores. Similarly, achieving CDAI LDA after 6 or more months of etanercept was associated with younger age at onset, higher baseline CDAI, shorter disease duration, and fewer comorbidities. CONCLUSIONS: These patients with older-onset RA and more comorbidities clinically improved with etanercept, but had lower odds of achieving CDAI LDA. Age of onset and number of comorbidities may be important in determining RA tumor necrosis factor inhibitor response.

Occurrence of Coxiellosis in ruminants and its associated risk factors.

Coxiellosis in animals is caused by the zoonotic pathogen, Coxiella burnetii. Although the disease is of public health importance it remains underdiagnosed and underreported. The cross- sectional study was aimed to estimate the occurrence of the disease in livestock of study area and also to identify the risk factors associated with the disease in animals. Blood, serum, and vaginal swabs samples were collected from 200 ruminants (cattle, sheep, and goats), across various farms in Karnataka, India. These samples were then screened using ELISA and PCR (com1 and IS1111). A questionnaire was administered to the farm owners to collect the risk factor-related information. About 5.26 % cattle, 12.3 % sheep, and 12.5 % goats were positive by ELISA. By PCR, 9.47 % cattle, 9.3 % sheep, and 10 % goats were positive. Overall, the occurrence of 14.73 %, 18.46 % and 17.5 % was estimated in cattle, sheep and goat, respectively. PCR targeting the IS1111 gene detected higher number of samples as positive as compared to the com1 gene PCR. Higher number of vaginal swab samples were detected as positive as compared to blood. History of reproductive disorders (OR: 4.30; 95 %CI:1.95- 9.46), abortion (OR: 30.94; 95 %CI:6.30- 151.84) and repeat breeding (OR:11.36; 95 %CI:4.16- 30.99) were significantly associated with coxiellosis (p < 0.005). Multivariable analysis by logistic regression model analysis suggested retained abortion, repeat breeding and rearing of animal in semi-intensive system as factors significantly associated with the infection. Cultural identification of the PCR positive samples were cultured using embryonated egg propagation and cell culture techniques and positivity was confirmed in six samples. Phylogenetic analysis of the com1 and IS1111 gene revealed clustering based on similar geographic locations. The study estimated the occurrence of the disease in the study area and identified the potential risk factors.

Comparative analysis of diagnostic assays for scrub typhus: Unveiling enhanced approaches for accurate detection.

The study comparatively evaluated serological assays, namely, Weil Felix assay, and IgM ELISA with the gold-standard immunofluorescence test (IFAT) for the sensitive and specific serodiagnosis of scrub typhus infection in occupationally exposed groups of humans. A total of 78 serum samples collected from persons affected with various ailments and belonging to different risk groups were screened in the study. Out of the 78 serum samples tested, a total of 17, 26, and 47 samples turned out to be positive by IFAT, IgM ELISA, and Weil Felix test, respectively. The Weil Felix assay could not serve as an ideal test for screening scrub typhus infection owing to its poor sensitivity and specificity in comparison with IFAT. IgM-ELISA could be an initial screening test to detect scrub typhus suspected patient in limited resource settings.

Comparison of physician and patient global assessments over time in patients with rheumatoid arthritis: a retrospective analysis from the RADIUS cohort.

BACKGROUND: In rheumatoid arthritis (RA), there is discordance between patient and physician assessments of disease severity and treatment response. OBJECTIVE: This retrospective analysis of the RADIUS (RA Disease-Modifying Anti-Rheumatic Drug Intervention and Utilization Study) 1 cohort examined specific factors that influence differences in global assessments for therapeutic effectiveness of disease-modifying antirheumatic drugs made by physicians (physician global assessment [PhGA]) and patients (patient global assessment [PtGA]). METHODS: The RADIUS 1 cohort consisted of primarily community-based private practice patients with RA requiring either the addition of or a switch to a new biologic or nonbiologic disease-modifying antirheumatic drug and who were followed for up to 5 years by their rheumatologists. Periodic assessments included PhGA, PtGA, Health Assessment Questionnaire-Disability Index (HAQ-DI), 28-item tender/painful joint count (TJC28), swollen joint count (SJC28), pain Visual Analog Scale (VAS), and acute-phase reactants. RESULTS: Among 4359 patients (mean disease duration, 7.3 years), PhGA most highly correlated with TJC28 (0.6956; 95% confidence interval [CI], 0.6881-0.7030) and SJC28 (0.6757; 95% CI, 0.6678-0.6834). Moderate overall correlations were observed for PtGA with TJC28 (0.5000; 95% CI, 0.4890-0.5108) and less 50 with SJC28 (0.3754; 95% CI, 0.3628-0.3878). Patient global assessment most strongly correlated with pain VAS (0.8349; 95% CI, 0.8305-0.8392) and moderately correlated with HAQ-DI (0.5979; 95% CI, 0.5886-0.6071). Acute-phase reactants poorly correlated with PhGA and PtGA. CONCLUSIONS: Low correlations between PhGA and acute-phase reactants suggest that these measurements have a limited contribution compared with the physical examination when physicians make global assessments. These results also suggest that physicians should consider patients' assessments of their disease activity (HAQ, pain VAS, and PtGA) and put joint counts into proper context.

Optimization of In-House Indirect-ELISA & EITB Assays Employing Cysticercus cellulosae Antigens for Serological Detection and PCR Assays for Molecular Detection of Porcine Cysticercosis.

Background: Porcine cysticercosis, caused by metacestodes of Taenia solium is an important neglected zoonosis. We evaluated the presence of anti-cysticercal antibodies and T. solium specific DNA in pig sera and blood samples respectively collected from Maharashtra, India. Methods: A total of three antigens (Scolex Antigen (SA), Membrane Body Antigen (MBA) and Excretory-Secretory Antigen (ESA)) were prepared from metacestodes of T. solium and employed in an in-house developed indirect-IgG ELISA for serological screening of 1000 porcine sera samples at Department of Veterinary Public Health, Nagpur Veterinary College, Maharashtra, India. The ELISA positive sera samples were subjected to EITB Assay for detection of immunodominant peptides. An effort has been made for molecular detection of porcine cysticercosis by PCR assay targeting large subunit rRNA gene of T. solium from blood samples of the corresponding ELISA-positive pigs. Results: The overall seroprevalence of porcine cysticercosis employing SA, MBA and ESA was 12.6%, 8.7% and 12.5% respectively. The lower and medium molecular weight peptides were the most frequently recognised in EITB assay. The numbers of bands recognised in EITB assay were observed to be proportionate with the corresponding ELISA O.D. values. An amplification product of 286 bp was observed in 22.98% (20/87), 30.35% (30/99) and 17.14% (12/70) of the sero-positive samples against SA, ESA and MBA respectively. Conclusion: EITB still remains the gold standard serodiagnosis test for cysticercosis. The inclusion of a greater number of positive samples and purification of antigens may improve the diagnostic efficacy of the tests.

Is Atrophic Nonunion a Misnomer - A Hospital-based Prospective Cross-Sectional Study.

Objective The present study was conducted to estimate histologically the proportion of avascularity of fracture ends in case of nonunion of long bones. Methods A total of 15 cases of established quiescent nonunion were operated according to the standard protocol and the fracture ends were evaluated histologically. The biopsied tissue was briefly fixed with formalin, embedded with paraffin (FFPE), and 5-micron sections were stained with hematoxylin and eosin according to standard protocols. Immunohistochemistry with anti-CD31 antibody (JC70A clone, DBS) was performed manually using standard protocols. Results All cases of quiescent nonunion were included; radiologically, 2 cases were oligotrophic, and 13 cases were of atrophic nonunion. A total of 20% of the patients were females, 40% were in the age group between 31and 40 years old, and, radiologically, all cases were of atrophic nonunion. All cases showed positivity for CD-31 on immunohistochemistry. The blood vessel density was category I in 13.33% of the cases and category II in 86.67% of the cases. Four cases presented with mild inflammation and two presented with moderate inflammation. The average vessel count was 10 per high power field in the age groups between 20 and 30, 31 and 40, and 41and 50 years old. The age group between 61 and 70 years old showed an average vessel count of 4 per high power field. The difference in the vessel counts of oligotrophic and atrophic nonunion was not significant. No correlation was observed in the density of vessel count and duration of nonunion Conclusion The nomenclature for the classification of nonunion into atrophic, oligotrophic, and hypertrophic needs revision. Our findings do not support that atrophic and oligotrophic nonunion are histologically different.

Seroprevalence and associated risk factors of Toxoplasma gondii and Neospora caninum infections in cattle in Central India.

Toxoplasma gondii and Neospora caninum are closely related cyst-forming parasites identified as important causes of reproductive failures in ruminants. While these parasites have been reported worldwide, seroprevalence and associated risk factors for cattle infections have not been determined in India. A total of 576 serum samples of cattle were analyzed for antibodies to T. gondii and N. caninum using enzyme-linked immunosorbent assay (ELISA), modified/Neospora agglutination test (MAT/NAT), and an indirect fluorescent antibody test (IFAT-tachyzoite and bradyzoite). Additionally, general information about cattle, movement of cats and dogs, the menace of rodents, management, and reproductive disorders were assessed to identify the potential risk factors. Overall, 32.9% (190/576) serum samples reacted positively to T. gondii and 24.8% (143/576) to N. caninum. The performance of the diagnostic tests showed excellent agreement between IFAT and ELISA (kappa [κ] = 0.98) and between MAT/NAT and ELISA (κ = 0.97). Combining both infections on avidity test, 94% sera had high-IgG avidity, and 3% had low-IgG avidity antibodies, indicating chronic infection in the majority of the cases. The identified risk factors (p < 0.05) for exposure to T. gondii were: increasing age (Odds Ratio [OR]: 2.02), movement of cat (OR: 4.8) and rodents (OR: 1.57) in the farm; and for N. caninum: increasing age (OR: 1.6), movement of dogs in the farm (OR: 2.07), drinking pond water (OR: 1.64) and abortion (OR: 1.82). These findings revealed that T. gondii and N. caninum infections are widespread in the study area and suggest conducting nationwide epidemiological studies owing to their economic importance.

Seroprevalence and risk factors of Toxoplasma gondii infection among veterinary personnel and abattoir workers in Central India.

Toxoplasmosis, caused by the protozoan parasite Toxoplasma gondii, is an important zoonotic infection. Veterinary personnel and abattoir workers are considered to be at a high risk of T. gondii infection owing to their occupational exposure. However, the association of T. gondii infection with occupational exposure to animals has not been determined in India. Hence, we analysed 139 and 126 blood samples of veterinary personnel and abattoir workers, respectively, for anti-T. gondii antibodies using enzyme-linked immunosorbent assay (ELISA), modified agglutination test (MAT) and indirect fluorescent antibody test (IFAT). The association of seroprevalence with sociodemographic profiles, work activities and dietary habits was determined in the study population. MAT, ELISA and IFAT results demonstrated nearly 46%, 48% and 47% seropositivity, respectively. MAT (kappa = 0.924) and IFAT (kappa = 0.962) results showed good agreement with ELISA results. Of the ELISA positive samples, 46% was copositive for IgG antibody, 1.5% for IgM antibody and 1.5% for both IgG and IgM antibodies. High IgG avidity was observed only in IgG+ IgM- and IgG+ IgM+ samples and not in IgM+ IgG- samples, indicating chronic T. gondii infection in most of the cases. Furthermore, multivariate analysis revealed that T. gondii seropositivity was associated with age > 30 years (odds ration [OR] = 1.992), cat at home (OR = 1.991), not wearing gloves (OR = 1.886), not wearing safety glasses (OR = 1.985) and contact with soil (OR = 1.695). These findings support the presence of a potentially significant association between T. gondii seropositivity and occupational exposure to animals.

Evaluation of recombinant leptospiral surface antigen (Lsa27) lipoprotein for serodiagnosis of human leptospirosis by latex agglutination test.

PURPOSE: Leptospirosis has wide clinical presentations often mimicking other illnesses, thus rapid and simple diagnostics will have facilitated the initial patient management and therapy compared to other inaccessible and laborious tests/assays. METHOD: In this study, the sensitized latex beads coated with purified recombinant outer membrane (OM)-leptospiral surface antigen (Lsa27) lipoprotein of pathogenic Leptospira was evaluated as a diagnostic antigen in latex agglutination test (LAT) for the detection of anti-leptospiral antibodies in the human sera. The prepared rLsa27 latex beads were evaluated with the confirmed microscopic agglutination test (MAT) reactive (at 1:50) Leptospira-specific positive (n = 42) and non-reactive negative (n = 80) sera from human cases suspected of leptospirosis with the history of pyrexia of unknown origin. RESULT: The results revealed the relative diagnostic sensitivity of 90.48 % (confidence interval (CI) at 95 % : 77.4-97.3 %) and diagnostic specificity of 91.35 % (CI at 95 %: 82.8-96.4 %), with an accuracy of 90.98 % (CI at 95 %: 84.44-95.41 %), and the kappa value of 0.8036 ±â€¯0.056 SE (CI at 95 %: 0.69-0.91) with a substantial agreement against gold standard serological MAT. CONCLUSION: The findings suggest that the rLsa27 protein-based LAT can be useful as a simple rapid screening diagnostic test for the detection of anti-leptospiral antibodies in the sera of humans. This rapid test can be complemented by other confirmatory diagnostics for the early detection of Leptospira antibodies which may in turn help in the prompt treatment and mitigates the public health problem at primary health care level.

In silico molecular docking and in vitro antimicrobial efficacy of phytochemicals against multi-drug-resistant enteroaggregative Escherichia coli and non-typhoidal Salmonella spp.

BACKGROUND: In the wake of emergence of antimicrobial resistance, bioactive phytochemical compounds are proving to be important therapeutic agents. The present study envisaged in silico molecular docking as well as in vitro antimicrobial efficacy screening of identified phytochemical ligands to the dispersin (aap) and outer membrane osmoporin (OmpC) domains of enteroaggregative Escherichia coli (EAEC) and non-typhoidal Salmonella spp. (NTS), respectively. MATERIALS AND METHODS: The evaluation of drug-likeness, molecular properties, and bioactivity of the identified phytocompounds (thymol, carvacrol, and cinnamaldehyde) was carried out using Swiss ADME, while Protox-II and StopTox servers were used to identify its toxicity. The in silico molecular docking of the phytochemical ligands with the protein motifs of dispersin (PDB ID: 2jvu) and outer membrane osmoporin (PDB ID: 3uu2) were carried out using AutoDock v.4.20. Further, the antimicrobial efficacy of these compounds against multi-drug resistant EAEC and NTS strains was determined by estimating the minimum inhibitory concentrations and minimum bactericidal concentrations. Subsequently, these phytochemicals were subjected to their safety (sheep and human erythrocytic haemolysis) as well as stability (cationic salts, and pH) assays. RESULTS: All the three identified phytochemicals ligands were found to be zero violators of Lipinski's rule of five and exhibited drug-likeness. The compounds tested were categorized as toxicity class-4 by Protox-II and were found to be non- cardiotoxic by StopTox. The docking studies employing 3D model of dispersin and ompC motifs with the identified phytochemical ligands exhibited good binding affinity. The identified phytochemical compounds were observed to be comparatively stable at different conditions (cationic salts, and pH); however, a concentration-dependent increase in the haemolytic assay was observed against sheep as well as human erythrocytes. CONCLUSIONS: In silico molecular docking studies provided useful insights to understand the interaction of phytochemical ligands with protein motifs of pathogen and should be used routinely before the wet screening of any phytochemicals for their antibacterial, stability, and safety aspects.

Evaluation of FKBP5 as a cortisol activity biomarker in patients with ACTH-dependent Cushing syndrome.

PURPOSE: To evaluate the performance of FKBP5 as a cortisol activity biomarker in patients with ACTH-dependent Cushing syndrome (CS). METHODS: This was a prospective, multicenter, nonrandomized, noninterventional study of a cortisol activity biomarker in adult patients (≥18 years) with documented ACTH-dependent, endogenous CS. The impact of surgery on FKBP5 mRNA expression levels in these patients and the difference in expression levels between these patients and healthy controls were evaluated. Cortisol and biomarker samples were collected before and immediately after surgery. A custom NanoString assay was used to quantify FKBP5 mRNA expression levels. The same method was used to analyze healthy volunteer samples collected from a different study. RESULTS: Surgery was considered successful in 14/24 patients (58.3%) and changes from baseline in serum cortisol were -92.6% (P = 0.0005) and -43.8% (not significant) in patients with successful and unsuccessful surgeries, respectively. A strong positive correlation between FKBP5 and cortisol levels was observed (before surgery: r = 0.72, P = 0.0002; after surgery: r = 0.85, P < 0.0001). After successful surgery, FKBP5 expression levels were similar to those of healthy subjects. In patients without surgical success, FKBP5 levels remained unchanged from baseline and distinct from healthy subjects (P = 0.0025). CONCLUSIONS: Our findings confirm that FKBP5 levels are higher in the presence of excess cortisol exposure in patients with CS and decrease to normal baseline levels after successful surgery. These findings suggest that FKBP5 can serve as a measure of biological cortisol activity and set the stage for the development of an FKBP5 mRNA expression assay as a biomarker of cortisol activity.