Pharmacokinetics of ceftriaxone-tazobactam (8:1) combination in healthy and Escherichia coli induced diarrhoeic birds.

Antibiotic-resistant Escherichia coli infection of poultry causes significant economic losses. Extended spectrum ß lactamases (ESBL) producing E. coli was inoculated in a broiler, Rhode Island Red and Haringhata Black birds orally at 56×108 c.f.u. mL-1 for induction of diarrhoea. Pharmacokinetics of ceftriaxone-tazobactam combination (8:1) was studied following a single intramuscular injection at 28.125 mg kg-1 and the combination was administered twice daily to treat such infection. Plasma concentration of both ceftriaxone persisted up to 8 h in experimental birds and maintained an approximate ratio of 8:1 with tazobactam for a period of 2 h, 0.25 h and 0.75 h, respectively in a broiler, Rhode Island Red and Haringhata Black birds. The K el was significantly lower in all experimental birds compared to healthy birds. Efficacy study was conducted in diarrhoeic birds by administration of ceftriaxone-tazobactam combination at 28.125 mg kg-1 body weight twice daily intramuscularly for three days which caused an increase in specific antibody titre in the broiler on 5th day and in Rhode Island Red birds 10th day. However, Haringhata black birds were inherently showed more resistance towards the infection. The combination of ceftriaxone and tazobactam in the ratio of 8:1 can be an effective treatment to combat ESBL producing E. coli infections.

Adrenocortical Carcinoma: a Therapeutic Challenge - 44 Cases from a Single Tertiary Care Center in India.

This study was conducted among patients with adrenocortical carcinoma (ACC) to analyze their clinico-pathological profile, management outcomes, and risk factors for local recurrence, systemic metastasis, and survival. The data of patients with ACC who were managed at a single institution between January 2004 and December 2016 was retrospectively collected and analyzed using STATA 13.1. Forty-four patients with a diagnosis of ACC were included in the study. The mean age at presentation was 38.5 ± 14.6 (9-74) with a male preponderance. Functioning tumors represented 59.1% (n = 26), cortisol being the most common hormone secreted. Forty patients (90.9%) underwent surgery, 14 (35%) of whom required an en bloc resection of adjacent organs. Fifteen (37.5%) received radiation (RT) to the postoperative bed while chemotherapy and mitotane were administered in 12 (27.3%) and 9 (20.5%) respectively. The mean follow-up was 34.3 ± 32.7 months. Twelve (30%) patients developed local recurrence, 21 (55.3%) had systemic metastasis, and 15 (34.1%) expired. The mean 1-year and 5-year overall survival rates were 77% and 65.7% respectively. On multivariate analysis, patients with ENSAT stage III/IV were significantly associated with local recurrence (p = 0.011) and metastasis (p = 0.037). Age > 50 (p = 0.003) and ENSAT III/IV (p = 0.017) were significantly associated with mortality on univariate analysis but not on multivariate analysis. In our study population, patients presented at a younger age with a male preponderance. Ninety percent underwent surgery, a subset (35%) requiring resection of adjacent organs to ensure R0 resection. Patients presenting at ENSAT stage I/II have better outcomes.

Rhamnolipid Biosurfactant against Fusarium verticillioides to Control Stalk and Ear Rot Disease of Maize.

Antifungal activity of rhamnolipids (RLs) has been widely studied against many plant pathogenic fungi, but not against Fusarium verticillioides, a major pathogen of maize (Zea mays L.). F. verticillioides causes stalk and ear rot of maize or asymptomatically colonizes the plant and ears resulting in moderate to heavy crop loss throughout the world. F. verticillioides produces fumonisin mycotoxins, reported carcinogens, which makes the contaminated ears unsuitable for consumption. In this study, the RL produced using glucose as sole carbon source was characterized by FTIR and LCMS analyses and its antifungal activity against F. verticillioides was evaluated in vitro on maize stalks and seeds. Further, the effect of RL on the mycelia of F. verticillioides was investigated by scanning electron microscopy which revealed visible damage to the mycelial structure as compared to control samples. In planta, treatment of maize seeds with a RL concentration of 50 mg l-1 resulted in improved biomass and fruiting compared to those of healthy control plants and complete suppression of characteristic disease symptoms and colonization of maize by F. verticillioides. The study highlights the potential of RLs to be used for an effective biocontrol strategy against colonization of maize plant by F. verticillioides.

Seroepidemiology of bluetongue in South Bengal.

AIM: With the aim of revealing the epidemiological intricacies of bluetongue (BT) in the southern part of West Bengal state, the present study was undertaken to assess seroprevalence of BT along with identification of the vector of the disease, i.e., Culicoides midges available in the region in their breeding season with conducive environmental factors, if any. MATERIALS AND METHODS: A total of 1509 (sheep-504, goat-1005) samples were collected from three different agroclimatic zones of South Bengal viz. new alluvial, red laterite and coastal saline. To detect anti-BT antibodies in the collected serum samples, indirect-enzyme-linked immunosorbent assay (i-ELISA) was performed. Culicoides midges were collected from those agro-climatic zones of South Bengal for species identification. The meteorological parameters, viz. temperature (maximum and minimum), rainfall and relative humidity of three agro-climatic zones of South Bengal were analyzed for the months of July to December during 2010-2013. RESULTS: The overall seropositivity was 33.13% and 30.24% in sheep and goat, respectively as assessed by i-ELISA. In South Bengal, the predominant species of Culicoides found were Culicoides schultzei, Culicoides palpifer and Culicoides definitus. CONCLUSION: Since virus transmitting species of Culicoides midges could be detected in South Bengal, besides high seropositivity in ruminants, the possibility of circulating BT virus in South Bengal is quite imminent.

Correlation of serum estradiol level on the day of ovulation trigger with the reproductive outcome of intracytoplasmic sperm injection.

BACKGROUND: Serum estradiol (E2) levels are measured in in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI), to assess the ovarian response and to predict ovarian hyperstimulation syndrome. The impact of peak E2 levels on IVF-ICSI outcome was found to be inconsistent in the previous studies. AIM: To evaluate the impact of the serum E2 levels on the day of ovulation trigger with the reproductive outcome of ICSI. SETTINGS AND DESIGN: Retrospective observational study. ART Center, at a Tertiary Care University Teaching Hospital. SUBJECTS AND METHODS: Eighty-nine infertile women, who underwent ICSI with fresh embryo transfer over a period of 3 years, were included in the study. The study subjects were grouped based on the serum E2 level on the day of ovulation trigger:- Group I - <1000 pg/ml, Group II - 1000-2000 pg/ml, Group III - 2000.1-3000 pg/ml, Group IV - 3000.1-4000 pg/ml, and Group V >4000 pg/ml. The baseline characteristics and controlled ovarian hyperstimulation (COH) outcome were compared among the study groups. STATISTICAL ANALYSIS USED: Chi-square test, Student's t-test, ANOVA, and logistic regression analysis. RESULTS: The study groups were comparable with regard to age, body mass index, ovarian reserve. Group V had significantly higher number of oocytes retrieved than I and II (18.90 vs. 11.36 and 11.33; P = 0.009). Group IV showed significantly higher fertilization rate than I, III, and V; (92.23 vs. 77.43, 75.52, 75.73; P = 0.028). There were no significant differences in the implantation rates (P = 0.368) and pregnancy rates (P = 0.368). CONCLUSION: Higher E2 levels on the day of ovulation trigger would predict increased oocyte yield after COH. E2 levels in the range of 3000-4000 pg/ml would probably predict increased fertilization and pregnancies in ICSI cycles.

Approaches to characterize extended spectrum beta-lactamase/beta-lactamase producing Escherichia coli in healthy organized vis-a-vis backyard farmed pigs in India.

The study was undertaken to investigate the occurrence and to characterize the ESBL/beta-lactamase producing-Escherichia coli in healthy pigs of organized and backyard farms in West Bengal, India. Total 200 rectal swabs were collected randomly from healthy pigs maintained in four organized farms and 10 backyard farms (n=100 each) and 76 isolates were identified as E. coli from organized (48/100, 48%) and backyard pigs (28/100, 28%). Twelve E. coli isolates (6%) in the present study were detected to possess any of the ESBL/beta-lactamase genes studied. ESBL/beta-lactamase producers were isolated with significantly more frequency from backyard pigs than the organized farm pigs (p=0.026). Six of ESBL/beta-lactamase producing isolates were phenotypically confirmed as CTX-M producers and ten of them were confirmed as TEM/SHV producers. PCR and sequencing of the amplified product from representative isolates revealed the presence of blaCTX-M-9, blaSHV-12 and blaTEM-1. No unique combination of the studied beta lactamase genes for organized and backyard farm pig isolates was noted. The ESBL isolates belonged to O13, O55, O133, O153, O157, O158, O166, rough and OUT serogroups. The association of heat labile toxin (elt) (p<0.0005) with organized farm isolates and heat stable toxin (estA) (p=0.0143) with backyard piggery sector was significantly higher. The ESBL/beta-lactamase producers from organized farm (Ak/Ex) and indigenous pigs (Ak/Ex/Te; Ak/CoT/G) showed a characteristic phenotypical antibiotic resistance pattern. Two pairs of isolates from organized and backyard farm pigs showed clonal relationship indicating a possible transmission between the farms which were situated adjacently. Thus the present study revealed backyard farm pigs as major source of ESBL/beta-lactamase producing-E. coli associated with STa and characteristic antibiotic resistance pattern in India.

Detection of T- and B-cell Target Antigens of Fowlpox Virus Isolated from Backyard Chickens in India.

With the aim of assessing the antigenic characteristics of a circulating pool of fowlpox virus (FPV) that exists in the backyard poultry system in India, one of the field isolates generated was characterized by in vitro immunologic techniques. FPV was isolated from clinically positive fowlpox cases (n  =  10) from the Jhargram (West Midnapur district) and Kakdwip (South 24 Pargana district) areas of West Bengal State, India. Initially, FPV-specific PCR was performed for confirmation of the samples. Isolation of FPV was done using embryonated chicken eggs and the choreoallantoic membrane route. Subsequently, FPV antigen was prepared from chicken embryo fibroblast cell culture-adapted field isolate. Biologic transmission of FPV was performed in Rhode Island red chickens experimentally to assess humoral and cell-mediated immune (CMI) responses. High level of anti-FPV antibodies were observed in test birds as assessed by indirect ELISA. Seroreactive polypeptides (B-cell antigens) of FPV antigen with molecular weights of 44.5, 66.5, 75, 90.5, and 99 kDa were detected by western blot analysis. Significant increases in CMI responses were observed in inoculated chickens as assessed by lymphocyte proliferation assay, cytotoxicity assay, and T-cell immunoblotting. The predominant T-cell antigen of FPV detected had a molecular weight of 66.5 kDa. The present study revealed the antigenic characteristics of FPV that exists in backyard poultry system in West Bengal for the first time, thus exploring the rationality of designing future T- and B-cell vaccines against fowlpox.

In vitro and in vivo evaluation of the marine sponge skeleton as a bone mimicking biomaterial.

This investigation was carried out to identify and characterize marine sponges as potential bioscaffolds in bone tissue engineering. The marine sponge (Biemna fortis) samples were collected from the rocky intertidal region of Anjuna, Goa, India, freeze-dried and converted to pure cristobalite at low temperature. After thorough evaluation of sponge samples by DTA-TGA thermography, XRD, FTIR, SEM and cell cytotoxicity by MTT assay, bare sponge scaffolds were fabricated by firing at 1190 °C. These scaffolds were loaded with growth factors (IGF-1 and BMP-2), checked for quasi-dynamic in vitro release kinetics and finally implanted into femoral bone defects in rabbits for up to 90 days, by keeping an empty defect as a control. The in vivo bone healing process was evaluated and compared using chronological radiology, histology, SEM and fluorochrome labeling studies. SEM revealed that the sponge skeleton possesses a collagenous fibrous network consisting of highly internetworked porosity in the size range of 10-220 µm. XRD and FTIR analysis showed a cristobalite phase with acicular crystals of high aspect ratio, and crystallinity was found to increase from 725 to 1190 °C. MTT assay demonstrated the non-cytotoxicity of the samples. A combination of burst and sustained release profile was noticed for both the growth factors and about 74.3% and 83% total release at day 28. In the radiological, histological, scanning electron microscopy and fluorochrome labeling analysis, the IGF-1 impregnated converted sponge scaffold promoted excellent osseous tissue formation followed by the BMP-2 loaded and bare one. These observations suggest that the marine sponge alone and in combination with growth factors is a promising biomaterial for bone repair and bone augmentation.

Virulence repertoire, characterization, and antibiotic resistance pattern analysis of Escherichia coli isolated from backyard layers and their environment in India.

This study was undertaken to observe the prevalence, serogroup, avian pathogenic Escherichia coli (APEC)-associated virulence gene, randomly amplified polymorphic DNA (RAPD) pattern, and antibiotic resistance genes of E. coli in backyard layers and their environment in India. From the 360 samples of healthy layers and their environment, 272 (75.5%) E. coli were isolated. The majority (28.67%) of them were untypeable. Among the studied virulence genes (papC, tsh, iucC, astA), 52 (14.32%) isolates were found to possess astA, including the isolates from the drinking water of the birds (4/272, 1.47%). These strains belonged to 18 different serogroups. Most of the isolates were typeable by RAPD and they produced different patterns. Phenotypic resistance of the isolates was most frequently observed to erythromycin (95.83%), chloramphenicol (87.52%), and cotrimoxazole (78.26%). None of the isolates was found to possess extended-spectrum beta-lactamases (bla(TEM), bla(SHV), bla(CTX-M) or quinolone resistance (qnrA) genes by PCR. The present study was the first attempt in India to assess APEC distribution in backyard poultry production.

Hypoglycaemic: prevention, consequences and management.

Hypoglycaemia is one of the most commonly encountered acute complications of diabetes. This is caused due to overdosage of insulin or oral hypolgycaemic agents. The signs and symptoms of hypoglycaemia can be divided into those due to autonomic hyperactivity and those resulting from neuroglycopenia. If detected early, hypoglycaemia is reversible and can be prevented. Hypoglycaemia may be misdiagnosed as hyperglycaemia and if not treated promptly may lead to death. The treatment of choice in hypoglycaemia is to give 25-50 ml of 50% glucose solution over 2-3 minutes. Where peripheral veins are collapsed, i.v. glucagon injection is preferable. Adequate education of diabetic patients and their relatives is necessary so that hypoglycaemia can be avoided or treated effectively.

Dynamic changes in cellular immune responses in experimental bovine tuberculosis.

BACKGROUND: The development of T-cell responses in natural hosts, such as cattle, would be a useful model of tuberculosis to investigate the changing pattern of T-cell responses and mycobacterial antigens recognized along with the progression of the disease. MATERIAL/METHODS: The present study was undertaken to characterize immune-effector cells and cytokines in Mycobacterium bovis AN5-sensitized cattle, from day 0 to 45 weeks post inoculation (PI). In vitro lymphoproliferation, cytotoxic T lymphocyte (CTL) response, and cytokine release pattern (viz. IL-2, IFN-gamma) from activated PBMC were assessed throughout the spectrum. The effort was made to identify various T-cell target antigens during different post sensitization stages, using T-cell immunoblotting. RESULTS: Cell-mediated immune (CMI) responses were very high between the 9th and 20th week PI. The early appearance of gd+ T-cells was followed by the predominance of CD4+ T-cells, and finally CD8+ T-cells until the 20th week PI. Lymphocyte-effector activity in terms of lymphoproliferation and CTL responses were quite evident between the 9th and 20th weeks PI and around the 12th week PI, respectively. Certain immunodominant polypeptides of culture filtrate preparation of molecular weights 72-74, 30-32, 25 & 22-24 kDa induced higher lymphoproliferation and CTL responses. The kinetics of in vitro cytokine release (IL-2 and IFN-gamma) from PBMC showed a definite trend in all M. bovis AN5-sensitized cattle, with a high concentration between the 9th and 20th weeks PI. CONCLUSIONS: The present investigation revealed some basic aspects of the immune-effector mechanism pertaining to bovine tuberculosis.