Anjana Goel

Trends and advances in the diagnosis and control of paratuberculosis in domestic livestock.

ABSTRACT Paratuberculosis (pTB) is a chronic granulomatous enteritis caused by Mycobacterium avium subsp. paratuberculosis (MAP) in a wide variety of domestic and wild animals. Control of pTB is difficult due to the lack of sensitive, efficacious and cost-effective diagnostics and marker vaccines. Microscopy, culture, and PCR have been used for the screening of MAP infection in animals for quite a long time. Besides, giving variable sensitivity and specificity, these tests have not been considered ideal for large-scale screening of domestic livestock. Serological tests like ELISA easily detects anti-MAP antibodies. However, it cannot differentiate between the vaccinated and infected animals. Nanotechnology-based diagnostic tests are underway to improve the sensitivity and specificity. Newer generation diagnostic tests based on recombinant MAP secretory proteins would open new paradigm for the differentiation between infected and vaccinated animals and for early detection of the infection. Due to higher seroreactivity of secretory proteins vis-à-vis cellular proteins, the secretory proteins may be used as marker vaccine, which may aid in the control of pTB infection in animals. Secretory proteins can be potentially used to develop future diagnostics, surveillance and monitoring of the disease progression in animals and the marker vaccine for the control and eradication of pTB.

Immunomodulating property of Ocimum sanctum by regulating the IL-2 production and its mRNA expression using rat's splenocytes

Objective: To investigate the regulatory effect of aqueous extract of leaves of Ocimum sanctum on IL-2 cytokine production in vivo and in vitro, and the effect of leaves extract on general blood picture including T& B lymphocytes. Methods: For in vivo studies albino Wistar rats were treated with aqueous crude leaves extract of Ocimum sanctum for 20 consecutive days. Spleen cells were harvested and assayed for IL-2 production by using sandwich enzyme-linked immunosorbent assay (ELISA) and mRNA expression methods. For in vitro study aqueous Ocimum sanctum leaves extract= in different concentrations (25-500 滋g/mL) was added into culture plates containing ConA stimulated splenocytes. To study the overall effect on blood picture, density gradient purified lymphocytes analysis and conventional methodology for total and differential leukocyte count and hemoglobin level were also done. Results: It indicated that the rats treated with Ocimum sanctum leaves extract had significantly enhanced (P<0.001) ability of spleen cells to secrete IL-2. Investigation in vitro also showed regulation of IL-2 production. Blood study exhibited leucocytosis and augmentation of T& B lymphocytes by 25% approximately. 4-5% increase in Hemoglobin value was also noticed. Conclusion: Aqueous Ocimum sanctum leaves extract may have stimulatory effect on T & B lymphocytes particularly on Th1 subset of lymphocytes as shown by enhancement in IL-2 production.

Effect of Ocimum Sanctum on the Development of Protective Immunity against Salmonella typhimurium Infection through Cytokines

To investigate the protective role of Ocimum sanctum (O. sanctum) leaves against Salmonella typhimurium (S. typhimurium) infection in rats by inducing TNF-α, IFN-γ & IL-2 cytokines. Methods: Wistar albino rats were fed with aqueous extract of O. sanctum leaves using 250mg/kg body weight dose once a day for 20 consecutive days. Control rats were fed with placebo. Rats were infected with LD50 dose of S. typhimurium infection and monitored for their survival. Bacterial blood burden in both the groups was compared and numbers of activated peritoneal macrophages were counted. Concentration of TNF-α, IFN-γ and IL-2 cytokines in serum during different time intervals was assayed by sandwich ELISA. Results: Rats of control group showed a high mortality rate and had higher bacterial blood burden when compared with O. sanctum extract fed rats. There was a significant increase in the number of S. typhimurium engulfed peritoneal macrophages in the peritoneal fluid of O. sanctum fed animals. The protective control against bacterial infection in O. sanctum fed rats was associated with elevated level of TNF-α, IFN-γ and IL-2 cytokines in serum. Conclusions: These findings suggest that orally administered O. sanctum leaves extract effectively enhanced activation in macrophage and lymphocytes, depicted by the elevated serum concentration of TNF-α, IFN-γ and IL-2 cytokines, leading to induce a protective resistance against Salmonella typhimurium infection.

Antiepileptic Effect of Nux vomica, Homeopathic Remedy, Against Strychnine-Induced Seizers

Objective: To investigate the antiepileptic effect of homeopathic remedy Nux vomica on mice and its comparison with standard therapeutic diazepam. Methods: BALB-c mice were taken and divided into three groups comprising ten mice in each group. The first group was treated as control; the second group received standard therapeutics (diazepam, i.p.) and the third group received Nux vomica CH7. All groups were treated with strychnine intra peritoneally. Following parameters were observed; start time of convulsions, the number of animals had convulsions, and survival time until death. Results: Nux vomica CH7 homeopathic preparation was found effective in suspending onset of convulsions (P˂ 0.01), and extending survival time until death (P˂ 0.01) in comparison to control mice. It also increased percentage survival in comparison to control as well as diazepam treated animals. Conclusion: Our study demonstrated efficacy of Nux vomica in epilepsy management.

Establishing the genetic variation in physiological response in response to heat stress in semi-arid region in Jamunapari goats

ABSTRACT The genetic variation in physiological response was analysed at population level in Jamunapari goats. The temperature humidity index (THI) range varied from 82.00 to 92.08 during hot period, 49.96 to 59.68 during cold period and 65.32 to 74.00 during thermo-neutral period. Physiological responses were recorded for 695, 617 and 291 animals during the hot, cold and thermo-neutral period, respectively. The sire mean of heat stress-susceptible (HSS) and heat stress-tolerant (HST) families differed by 19.737 in respiration rate (RR) and 31.438 in heart rate (HR). RR and HR were significantly different (P < 0.01) between HSS and HST phenotype. However, rectal temperature (RT) did not differ significantly (P < 0.01) between HSS and HST phenotype. The heritability of RT, RR and HR were 0.180 ± 0.058, 0.077 ± 0.044 and 0.307 ± 0.076, respectively. Genetic correlation between RT and HR was high during hot and cold period. The genetic correlation between RT and HR was high and positive during hot and cold period. The phenotypic correlation between RT and HR ranged from 0.132 to 0.190. The genetic correlation between RT with other traits was low and negative. Our finding indicates that phenotypic difference can be heritable.

Immunomodulating potential of Argemone mexicana through cytokine regulation

Biofilm formation on indwelling urinary catheters is a leading cause of Urinary tract infection (UTI). Presence of biofilm is associated with increased bacterial resistance to antimicrobial therapy and resultant treatment failure. The study detected a reliable method for diagnosis of biofilm formation by comparing scanning electron microscopy (SEM) and tissue culture plate method (TCP). The work was conducted on 20 urinary catheters from patients ranging from 1.5 to 85 years with catheters that remained in situ for a period of 3 to 20 days. Samples of catheters for culture and SEM and samples of urine were taken at the same time. The correlation between renal conditions and biofilm formation was not significant (p=0.336). No significant correlation (p =0.836, 0.163 respectively) was found between predisposing conditions (DM, renal insufficiency, diarrhea and impaired immunity) and development of Catheter associated urinary tract infection ( CAUTI )and biofilm formation. Biofilm formation increased with duration of catheter in situ, but no significant.correlation was found (p=0.095). This could be due to small number of specimens. 9/20(45%) urine samples, 12/20(60%) catheter samples were positive by culture and 14/20(70%) catheters showed biofilm on SEM. 4/12(33.33%) organisms isolated from catheter culture produced biofilm by TCP method. 9 isolates were recovered from 9 positive urine cultures. The microorganisms isolated were non Candida albicans (3/9), E. coli (2/9), C. albicans (2/9) and Acenitobacter (2/9). 14 isolates were recovered from 12 culture- positive catheters. The organisms isolated were E.- coli (3/14), non-Candida albicans (3/14), C. albicans (2/14), C tropicalis (2/14), Acenitobacter (2/14), Klebsiella (1/14) and Enterococcus (1/14). Reduction in microbial diversity with antimicrobial use was noticed but the correlation was insignificant (p=0.317). The correlation between urine culture results as well as catheter culture results and biofilm formation by SEM were both significant (p = 0.008 & 0.000 respectively). The correlation between urine culture and TCP assay was insignificant (p =0.237). Using SEM as the gold standard method for the detection of biofilm, the sensitivity, specificity; total accuracy, PPV & NPV of urine culture and catheter culture were, 64.30%, 100%, 75%, 100%, 54% & 85.70%, 100%, 90%, 100%, 75% respectively.

High Risk Human Papilloma Virus (HR-HPV) Genotyping in Female with Abnormal Cervix

Human Papilloma Virus (HPV) infection is transmitted primarily via sexual and affects mostly basal epithelial cells of the skin or inner lining of tissues and are categorized as cutaneous types or mucosal types. It is known that HPV infection is strongly related to the development of cervical cancer, as viral DNA is detected in 99% of cervical cancer cases worldwide. Proposed study includes 14 cervical brushing, collected from females with cytologically abnormal cervix. The target genes E6 and E7 were amplified by Conventional PCR. Three cases with High grade squamous intraepithelial lesion (HSIL) came positive. Further the positive cases were digested by restriction endonucleases enzymes for RFLP and the amplicon size for high risk HPV type 16 and type 18 were obtained at 452bp and 455bp respectively. Three cases came positive, out of which HPV type 16 came in two cases with one HPV type 18. High risk HPV characterization at an early stage will prevent further progression of the disease.