Mechanistic and recent updates in nano-bioremediation for developing green technology to alleviate agricultural contaminants.

The rise in environmental pollutant levels in recent years is mostly attributable to anthropogenic activities such as industrial, agricultural and other activities. Additionally, these activities may produce excessive levels of dangerous toxicants such as heavy metals, organic pollutants including pesticide and herbicide chemicals, and sewage discharges from residential and commercial sources. With a focus on environmentally friendly, sustainable technology, new technologies such as combined process of nanotechnology and bioremediation are urgently needed to accelerate the cost-effective remediation process to alleviate toxic contaminants than the conventional remediation methods. Numerous studies have shown that nanoparticles possess special qualities including improved catalysis and adsorption as well as increased reactivity. Currently, microorganisms and their extracts are being used as promising, environmentally friendly catalysts for engineered nanomaterial. In the long term, this combination of both technologies called nano-bioremediation may significantly alter the field of environmental remediation since it is more intelligent, safe, environmentally friendly, economical and green. This review provides an overview of soil and water remediation techniques as well as the use of nano-bioremediation, which is made from various living organisms. Additionally, current developments related to the mechanism, model and kinetic studies for remediation of agricultural contaminants have been discussed.

Corticosteroid-associated lupus pancreatitis: a case series and systematic review of the literature.

BACKGROUND: Acute pancreatitis is an uncommon complication that occurs in 0.85% to 4% of patients with systemic lupus erythematosus (SLE). In some patients, it occurs within days to weeks of starting medium-to-high dose corticosteroids. The authors have used the term 'corticosteroid-associated lupus pancreatitis' for these patients, and they report a case series and perform a systematic review of previously published reports. METHODS: For the purpose of this study, corticosteroid-associated lupus pancreatitis was defined as occurrence of acute pancreatitis in patients with SLE (fulfilling the 1997 ACR), within 3 weeks of starting therapy with medium-to-high dose corticosteroids - either newly initiated or escalated from a lower dose. All patients with SLE admitted in the last 2.5 years in a North Indian university hospital were reviewed, and those with pancreatitis who fulfilled the above criteria were included in the case series. For the systematic review, a PUBMED search using the keywords 'lupus' and 'pancreatitis' was performed, and reports in English were reviewed for an association with corticosteroids. RESULTS: Among 420 admissions of SLE patients, six patients (1.4%) fulfilled criteria for corticosteroid-associated lupus pancreatitis. All were female, with mean age and disease duration of 19.7 ± 3.3 and 3.8 ± 2.5 years respectively. All had active disease and developed acute pancreatitis within 48-72 hours of newly initiating medium-to-high dose corticosteroids (in three patients) or escalating them to medium-high dose (in three patients). After the development of pancreatitis, corticosteroids were continued in all except one patient. In addition, two patients received pulse methylprednisolone, two received pulse cyclophosphamide and one was started on azathioprine. Three patients died during hospitalization, all with severe pancreatitis. On systematic review, among 451 cases of lupus pancreatitis reported, 23 (5%) fulfilled criteria for 'corticosteroid-associated lupus pancreatitis'. A majority of them had pancreatitis within 3 days of starting treatment with medium-to-high dose corticosteroids. The mortality in these patients was 37.5%. CONCLUSION: In a small but substantial proportion of patients with lupus who develop pancreatitis, it occurs within days to weeks of starting medium-to-high dose corticosteroids. Many of these patients continue to receive corticosteroids, and some receive more aggressive immunosuppression. However, they have significant mortality, and further studies are required to identify appropriate treatment in this subgroup of patients.

Phylogenetic analysis of VP2 gene of canine parvovirus and comparison with Indian and world isolates.

Canine parvovirus (CPV) causes hemorrhagic enteritis, especially in young dogs, leading to high morbidity and mortality. It has four main antigenic types CPV-2, CPV-2a, CPV-2b and CPV-2c. Virus protein 2 (VP2) is the main capsid protein and mutations affecting VP2 gene are responsible for the evolution of various antigenic types of CPV. Full length VP2 gene from field isolates was amplified and cloned for sequence analysis. The sequences were submitted to the GenBank and were assigned Acc. Nos., viz. KP406928.1 for P12, KP406927.1 for P15, KP406930.1 for P32, KP406926.1 for Megavac-6 and KP406929.1 for NobivacDHPPi. Phylogenetic analysis indicated that the samples were forming a separate clad with vaccine strains. When the samples were compared with the world and Indian isolates, it was observed that samples formed a separate node indicating regional genetic variation in CPV.

Subclinical circulation of Chicken infectious anaemia virus- A Sero-molecular study.

Chicken infectious anaemia-an important immunosuppressive viral disease of chicken-gained much attention in the recent past. Based on huge mortality and production loss observed in the fast-growing poultry sector, the present study aimed to find out the current status of the chicken infectious anaemia virus (CIAV), among chicken flocks in the Punjab state of India by sero-molecular study. The sera from the blood samples were tested for anti-CIAV antibodies by indirect ELISA and also compared with haematological parameters. DNA from sero-positive samples underwent PCR amplification, sequencing and phylogenetic analysis of the most conserved genomic region (VP3 gene) to detect viraemia in asymptomatic birds. The serological study using indirect ELISA showed a high sero-positivity of 77.27% in chicken flocks. Additionally, the present study also revealed the high molecular evidence (72.54%) of CIAV in apparently healthy birds. Genetic analysis showed that all CIAVs have conserved VP3 genes without any nucleotide substitutions, indicating presence of CIAV and its subclinical circulation among apparently healthy flocks. The wide distribution of CIAV among birds may be the reason for huge mortality and production loss. Further, it is suggested that studies be conducted to find out the co-involvement of CIAV with other immunosuppressive microbial agents and the immunosuppressive effect of CIAV in apparently healthy birds. Also, its role in vaccine failure and outbreaks of various other avian diseases needs to be explored.

Non-specific sex-differential effect of DTP vaccination may partially explain the excess girl child mortality in Ballabgarh, India.

OBJECTIVE: To test the hypothesis that a gender differential exists in the effect on child mortality of BCG, DTP, measles vaccine as administered under programme conditions in Ballabgarh HDSS area. METHODS: All live births in 28 villages of Ballabgarh block in North India from 2006 to 2011 were followed until 31 December 2011 or 36 months of age whichever was earlier. The period of analysis was divided into four time periods based on eligibility for vaccines under the national immunisation schedule (BCG for tuberculosis, primary and booster doses of diphtheria-tetanus-pertussis and measles). Cox proportional hazards regression was used to assess the association between sex and risk of mortality by vaccination status using age as the timescale in survival analysis and adjusting for wealth index, access to health care, the presence of a health facility in the village, parental education, type of family, birth order of the child and year of birth. RESULTS: 702 deaths (332 boys and 370 girls) occurred among 12,142 children in the cohort in the 3 years of follow-up giving a cumulative mortality rate of 57.5 per 1000 live births with 35% excess girl child mortality. Age at vaccination for the four vaccines did not differ by sex. There was significant excess mortality among girls after immunisation with DTP, for both primary (HR 1.65; 95% CI:1.17-2.32) and DTPb (2.21; 1.24-3.93) vaccinations. No significant excess morality among girls was noted after exposure to BCG 1.06 (0.67-1.67) or measles 1.34 (0.85-2.12) vaccine. CONCLUSION: This study supports the contention that DTP vaccination is partially responsible for higher mortality among girls in this study population.

Boning up on glypicans--opportunities for new insights into bone biology.

Bone formation is remarkable for the convergence in the activity of four major signalling pathways, the bone morphogenetic protein (BMP), fibroblast growth factor (FGF), hedgehog (HH) and wingless-integrated (WNT) pathways. These pathways cooperate in morphogenetic, proliferative and differentiative processes that underpin the development, growth and repair of skeletal structures. They are regulated by pathway-specific modulators and by another class of molecules, the glypicans. Glypicans are proteoglycans located on the cell surface, where they act as coreceptors to promote or inhibit signalling by ligands of the BMP, FGF, HH and WNT pathways, through protein-protein and protein-carbohydrate interactions. In this review, we discuss glypican structure, expression and function in the context of bone development and growth, with emphasis on the long bone growth plate where five of the six glypicans are expressed in overlapping patterns in the chondrogenic zone. Analyses of gene knockout models and the human conditions of Simpson-Golabi-Behmel syndrome and omodysplasia, which arise from mutations in glypican 3 (GPC3) and GPC6, respectively, highlight both subtle and striking effects of glypicans on bone growth. We draw attention to challenges and areas of opportunity, where the actions of glypicans on BMP, FGF, HH and WNT signalling might be profitably studied to help illuminate the complex interplay of signalling that drives bone growth.

Applicability of new sustainable and efficient green metal-based nanoparticles for removal of Cr(VI): Adsorption anti-microbial, and DFT studies.

Artemisia absinthium leaves were utilized as a reducing agent for green synthesis of Zinc oxide nanoparticles (particle size 17 nm). Synthesized green-ZnO (g-ZnO) were characterized by SEM/EDX, FTIR, XRD, UV, and BET analyses and then further used as an adsorbent to remove Cr(VI) ions from simulated wastewater. Optimal pH, temperature and adsorbent dosage were determined through batch mode studies. High removal efficiency and adsorption capacity were observed at pH 4, 0.25 g L-1 dosage, and 25 mg L-1 concentration of Cr(VI). Experimental data were modelled with different adsorption kinetics (Elovich model, PFO, PSO, IDP model) and isotherms (Langmuir, Freundlich, and Temkin), and it was found the adsorption process was well fitted to Langmuir with an R2 value greater than>0.99. Computational calculation showed that the g-ZnO nanoparticles became ∼14 times more dynamic with delocalized surface states making them a relevant platform to adsorb Cr with greater work function compatibility supporting the experimental findings. The Qmax adsorption capacity of g-ZnO was 315.46 mg g-1 from Langmuir calculations. Thermodynamic calculations reveal that the Cr (VI) adsorption process was spontaneous and endothermic, with a positive ΔS value representing the disorder at the solid-solution interface during the adsorption. In addition, the present study has demonstrated that these g-ZnO nanoparticles show strong antibacterial activities against P. aeruginosa (MTCC 1688) and E. coli (MTCC 1687). Also, the novel g-ZnO adsorbent capacity to remove Cr(VI) from simulated water revealed that it could be reused at least six times with higher removal rates during regeneration experiments. The results obtained from adsorption and antimicrobial activities suggest that g-ZnO nanoparticles could be used effectively in real-time wastewater and agricultural safety applications.

Fetomaternal outcome in pregnancy with severe thrombocytopenia.

OBJECTIVES: STo study the feto-maternal outcome in pregnancy with severe thrombocytopenia. MATERIALS AND METHODS: It was an observational study involving 1150 pregnant women with term gestation in labour, who were screened for thrombocytopenia. Ninety-four subjects (8.17%) were found to have thrombocytopenia i.e. platelet count < 1,50,/000/mm3, out of which 47 subjects (group A) had platelet count of less than 50.000/mm3 Simultaneously, 47 term pregnant women (group B) having a normal platelet count i.e. > 1.5 lac/mm3 formed the control group. All the subjects were followed during labour and postpartum period for any feto-maternal outcome. RESULTS: Significant history of bleeding tendencies like prolonged bleeding from wound site, easy bruisability and menorrhagia (p = 0.023) was evident in the study population. Abruptio placentae and early onset pregnancy induced hypertension (PIH) in previous gestations was more commonly found in the study population. Anemia and PIH were significantly more in group A. Incision site oozing during cesarean section was significantly more in group A. Moderate thrombocytopenia was more in neonates of study group (p = 0.014), but without any bleeding complications in neonates. CONCLUSIONS: Careful surveillance is required in these high risk patients for earlier detection and treatment of complications so as to decrease the fetomaternal morbidities.

Potential allergens of green gram (Vigna radiata L. Millsp) identified as members of cupin superfamily and seed albumin.

BACKGROUND: No systematic study on allergenicity of green gram seed proteins have been performed so far, although incidences of IgE-mediated reaction to green gram seedlings have been reported. OBJECTIVE: We sought to investigate the allergenic potential of green gram, followed by identification and characterization of its relevant allergens using proteomic approaches. METHODS: BALB/c mice were sensitized intraperitoneally with green gram proteins, and levels of specific Igs, Th2 cytokines, histamine, anaphylactic symptoms and histopathological responses were studied. Twelve naso-bronchial allergic patients with a history of sensitization to green gram were selected on the basis of positive skin prick test and elevated specific IgE levels. Green gram allergens were identified and characterized by their ability to endure pepsin, by IgE immunoblot of two-dimensional (2D) gels in combination with mass spectrometry and by bioinformatics approaches. RESULTS: Increased specific IgE, IgG1, Th2 cytokine and histamine levels, high anaphylactic scores and histological changes in lungs and spleen of green gram crude protein extract-treated mice are indicative of its sensitization ability. Four proteins (molecular weights: 52, 50, 30 and 18 kDa) showed pepsin resistance and IgE-binding capability with sensitized human and mice sera. The four proteins tentatively named as Vig r2 (52 kDa, pI 5.7), Vig r3 (50 kDa, pI 5.8), Vig r4 (30 kDa, pI 6.6) and Vig r5 (18 kDa, pI 5.5) showed significant sequence similarity with known allergens of soybean, lentil, pea, lupin, etc. Mass spectrometric analysis identified Vig r2 as 8S globulin ß-isoform precursor, Vig r3 as 8S globulin α-isoform precursor and Vig r4 as seed albumin. CONCLUSION AND CLINICAL RELEVANCE: Green gram seeds contain at least four clinically relevant allergenic proteins, namely Vig r2, Vig r3, Vig r4 and Vig r5 that were capable of inducing strong IgE-mediated reactions. One of the most important steps towards diagnostic and therapeutic approaches to deal effectively with food allergy is continued identification of newer food allergens and their characterization. The significance of this study can be enormous as the data generated may work as basic biology data in developing a green gram species modified genetically that may have reduced allergenicity.

Culture-free detection and enumeration of STEC in water.

Shiga toxin-producing Escherichia coli (STEC) causes worldwide outbreaks of food and waterborne diseases. Rapid identification of causative agents is critical for early intervention in the case of widespread diarrheal epidemics to prevent mortality. In this study, a Molecular-Beacon targeting stx2 gene (highly associated with human illness) was designed to develop a culture-independent real-time PCR assay for detection and quantification of STEC in water samples. The assay could detect lowest 10 genomic equivalent (GE) of the reference strain (E. coli I.T.R.C.-18) per PCR or 100 GE/mL. The presence of 10(6)CFU/mL of non-pathogenic E. coli DH5α has no impact on sensitivity of the assay. The assay could successfully enumerate STEC in surface water (collected from a sewage impacted river) and potable water samples collected from Lucknow city without prior enrichment. The assay will be useful in pre-emptive monitoring of surface/potable waters to prevent waterborne outbreaks caused by STEC.

Prevalence and molecular characterization of canine parvovirus.

BACKGROUND AND AIM: Canine parvovirus (CPV) belonging to family Parvoviridae causes hemorrhagic gastroenteritis in dogs and heavy mortality in young dogs. The virus has three structural (VP1, VP2 and VP3) and two non-structural proteins (NS1 and NS2), VP2 being highly immunogenic. This study aims to study molecular epidemiology of CPV by sequence analysis of VP2 gene to determine the prevailing antigenic type(s) in the northern regions of India. MATERIALS AND METHODS: A total of 118 rectal swabs collected from dogs exhibiting clinical signs of CPV infection were processed for the isolation of DNA and subjected to polymerase chain reaction (PCR) and nested PCR (NPCR). A total of 13 NPCR products selected randomly were subjected to sequence analysis of VP2 gene. RESULTS: The percent positivity of CPV was found 28% and 70% by PCR and NPCR, respectively. Dogs with vaccination history against CPV too were found positive with a percent positivity of 24.10%. Gene sequencing and phylogenetic analysis of VP2 gene from these isolates revealed that most samples formed a clade with CPV-2a isolates. CONCLUSION: Sequence analysis and phylogenetic analysis of VP2 gene in the studied regions of northern India revealed that CPV-2a was the most prevalent antigenic type.

Detection and molecular characterization of chicken infectious anaemia virus in young chicks in Punjab region of north-western India.

Chicken infectious anaemia (CIA) is an important viral disease of chicken causing significant immunosuppression and severe anaemia worldwide. Occurrence of severe disease and mortality is noticed in young chicks (2-3 weeks). Vertical mode of transmission increases chance of infection and persistence of virus among the infected flocks. The current study was conducted in Punjab state for confirmation and genetic characterization of CAV among chicken flocks of various poultry farms. DNA was extracted from the tissue samples and subjected to polymerase chain reaction (PCR) of VP1 gene and whole genome. PCR products were further sequenced for confirmation of chicken infectious anaemia virus (CIAV) genome in the clinical samples. PCR amplification of DNA from the tissue samples yielded expected product size of 1350 bases of VP1 gene and 2.3 kb of whole genome. Out of 16 commercial poultry farms, 11 were confirmed with presence of CIAV, and out of 65 birds, 39 were found positive (60%) for CIAV genes. Among the various organs, the presence of viral gene was detected at highest level in thymus when compared with other organs. It is concluded that chicken infectious anaemia virus detected from Punjab state is closely related to other Indian isolates and neighbouring countries which necessitates need of more intensive studies with a greater number of samples for implementing effective control measures.

Oral Candida infection and colonization in solid organ transplant recipients.

INTRODUCTION: Oral Candida carriage and infection have been reported to be associated with a greater risk for systemic infection in transplant recipients; however, a systematic analysis of the oral Candida titers and species has not been previously conducted. The objectives of this study were to determine the prevalence of oropharyngeal candidiasis, the oral carrier status, Candida titers and species in this population. METHODS: Ninety kidney and heart transplant subjects and 72 age-matched healthy controls were included. Swabs from the oral mucosa and a standardized amount of unstimulated saliva were plated on Chromagar Candida, and colony-forming units per millilitre were calculated. Initial speciation was based on colony color and was confirmed by standard germ tube, biotyping, or polymerase chain reaction assays. RESULTS: Infection with C. albicans was detected in seven transplant subjects and none of the controls. The transplant group had significantly higher oral Candida titers than the control group. There were no statistically significant relationships between the dose or type of immunosuppressants and oral Candida titers or infection. A significantly higher percentage of transplant subjects were colonized by more than one species, compared with control subjects. The most frequent species combination in transplant subjects was C. albicans and C. glabrata. C. glabrata was isolated from 13.5% of transplant carriers and none of the controls. CONCLUSIONS: Increased oral Candida infection and carriage titers were found in the transplant population. Although the majority of transplant patients were colonized by C. albicans, C. glabrata appears to emerge as the second most prevalent species.

Comparison of three techniques for the diagnosis of urinary tract infections in dogs with urolithiasis.

OBJECTIVES: To identify an appropriate sampling technique(s) to accurately detect the bacteria causing urinary tract infections in dogs with urolithiasis. METHODS: Twenty-one dogs with urolithiasis were included in the study. Three types of samples were taken from each dog. Urine was collected by cystocentesis, and a urinary bladder mucosal biopsy and urolith were retrieved during cystotomy. The samples were then cultured on blood agar and MacConkey's agar to identify the bacteria associated with urinary tract infections. RESULTS: Bacterial urinary tract infection was found in 16 cases (76.19 per cent). The most prevalent bacteria found to cause urinary tract infection were Escherichia coli (n=7), followed by coagulase-positive Staphylococcus species (n=4), Klebsiella pneumoniae (n=2), Pseudomonas aeruginosa (n=2) and Proteus mirabilis (n=1). In the case of a positive urine culture, the same bacteria were also cultured from the urinary bladder mucosal biopsy alone or from both the urinary bladder mucosal biopsy and urolith. However, in the case of a negative urine culture, bacteria were found to be present in the urinary bladder mucosal biopsy or urolith cultures in 23.81 per cent of dogs. The uroliths that gave positive culture results were either infection-induced uroliths composed of struvite and calcium carbonate phosphate, ammonium acid urate only or metabolic uroliths composed of calcium oxalate and calcium phosphate, or calcium phosphate only. All the uroliths that gave negative culture results were metabolic uroliths composed of calcium oxalate and/or calcium phosphate, and uric acid and calcium phosphate. CLINICAL SIGNIFICANCE: When the culture from the urine obtained by cystocentesis is negative, cultures of urinary bladder mucosal biopsy and urolith are recommended in dogs with urolithiasis in order to accurately assess the microbiological status of the urinary tract.

Multiplex real-time PCR for identification of canine parvovirus antigenic types.

Canine parvovirus (CPV) is an important disease causing gastroenteritis and/or haemorrhagic gastroenteritis in dogs. There are four antigenic types of CPV reported worldwide viz. CPV 2, CPV 2a, CPV 2b and CPV 2c. The diagnosis of CPV with the identification of the antigen type responsible remains problematic. In the present study, identification as well as antigenic typing of CPV was done using a de novo multiplex real time PCR to combat the problem of antigenic type identification. From the study it could be concluded that the here developed multiplex real time PCR assay could be used for rapid detection of CPV as well as typing of its three antigenic types.

Cumulative Risk of Metabolic Syndrome Correlated with the Coexistence of (-1306C/T) and Altered Circulating MMP2 level.

BACKGROUND: Interindividual genetic variations and environmental factors both play pivotal roles in the pathogenesis of metabolic syndrome (MetS). The rationale of this study conducted was to analyze the association of Matrix Metalloproteinase (MMP) gene variants, MMP-1 (-1607 1G/2G) and MMP-2 (-1306 C/T) with susceptibility to MetS and its effect on serum MMP level. METHODS: Study involved 370 subjects with 1:1 distribution of cases and controls. Patients were recruited according to modified NCEP-ATP III criteria for MetS. Clinical, biochemical analysis, PCR-RFLP and ELISA methods were employed for genotyping and estimation of serum MMP level. RESULTS: Significantly (p<0.001) higher Serum MMP-2 (39.13±19.96 ng/ml) was detected in cases as compared to controls. The MMP-2 (-1306 C/T) was significantly associated with the risk of MetS. The variant genotype TT was significantly associated with increased risk of MetS. (p=0.032; OR=2.31; 95%CI=1.07-4.97). No significant association of MMP-1(-1607 1G/2G) was found with risk of MetS. CONCLUSION: Our study concluded that presence of MMP-2 (-1306 C/T) might be associated the risk of MetS. Serum MMP2 level was significantly higher in patients and correlated with clinical parameters of MetS. Clinical implication of the work may help to identify the individuals with high risk of MetS and further complications.

Planar omnidirectional reflectors in chalcogenide glass and polymer.

We have fabricated and tested planar reflectors exhibiting an omnidirectional stop band centered near 1750 nm wavelength. The reflectors are comprised of multiple layers of Ge33As12Se55 chalcogenide glass and polyamide-imide polymer. Glass layers were deposited by thermal evaporation and polymer layers were deposited by spin-casting. Thin film stacks of up to 13 layers showed good planarity and adhesion, which we attribute to the well-matched thermo-mechanical properties of the materials. The optical properties of the reflectors were tested in both transmission and reflection, and the results are in good agreement with theoretical predictions. Relatively low-temperature processing steps were employed, making these reflectors of interest for integrated optics.

Teratogenic effects in rabbits of simultaneous exposure to ochratoxin A and aflatoxin B1 with special reference to microscopic effects.

To elucidate the teratogenic effects, ochratoxin A (OTA) and aflatoxin B1 (AFB1) were dissolved in corn oil and administered in combination to New Zealand White rabbits during 6-18 days of gestation orally with the dose levels of OTA+AFB1, 0.05+0.05 and 0.1+0.1mg/kg body weight. To assess pathomorphological features of the anomalies, the fetal serial sections were histologically examined. There was no mortality in any of the treated groups. Body weights and body weight gains of dams in the combined treatment groups were comparable with those of controls and individual treatments. The mean crown to rump lengths in both the combination dose groups and mean fetal weights in high dose combination group were significantly decreased. In the high dose combination, there was increase in the percent of implants resorbed and significant increase in the incidence of visceral anomalies. The combination treatment resulted in various gross, skeletal and visceral anomalies such as wrist drop, scoliosis, bent metacarpals, rudimentary ribs, cardiac defects and microphthalmia. There was a dose-related increase in the percent of litters showing the histopathological changes in the fetal tissues. The incidence of histopathological changes in the tissue sections prepared from fetal liver, kidneys, brain, heart and eyes was found increased in the high dose combination group. The comparative evaluation of the results of combination versus individual treatments revealed that certain anomalies observed in the individual treatment of OTA such as knuckling of fetlock, rudimentary tail or agenesis of tail, wavy ribs, hydrocephalus and agenesis of kidney and AFB1 as enlarged eye sockets and enlarged liver were absent in the combination treatment. However, some new manifestations such as cardiac defects and scoliosis were seen. The results of the present study indicated that in combination, OTA and AFB1 have antagonistic interaction. The presence of subtle lesions histologically due to an interference with normal development suggested that microscopic examination of the fetal tissues could provide additional, useful information to a developmental toxicity study.

Effects of aflatoxin B1 on embryo fetal development in rabbits.

Aflatoxin B1 (AFB1), is a food borne mycotoxin produced by fungal species of the genera Aspergillus. To elucidate the teratogenic effects, AFB1 was dissolved in corn oil and given orally to New Zealand White rabbits during 6-18 days of gestation with the dose levels of 0.025, 0.05 and 0.1 mg/kg body weight. To assess pathomorphological features of the anomalies induced by AFB1, the fetal serial sections were histologically examined. There was no maternal mortality in any group. There was non-significant decrease in percent of live fetuses and increase in the percent resorptions and post-implantation losses at 0.1 mg/kg dose group as compared with those of controls. The mean crown to rump lengths of 0.05 and 0.1 mg/kg dose groups were significantly reduced than that of the control. The mean fetal weights were significantly reduced in 0.1 mg/kg dose group than that of other treated groups. The gross anomalies observed included wrist drop and enlarged eye socket whereas, skeletal anomalies were agenesis of caudal vertebrae, incomplete ossification of skull bones and bent metacarpals. The visceral anomalies of microphthalmia and cardiac defects were seen at 0.1 mg/kg dose group. The characteristic histological findings of fetal tissues were distortion of normal hepatic cord pattern and reduced megakaryocytes in liver, fusion of auriculo-ventricular valves, mild degenerative changes in myocardial fibers, microphthalmic eyes and lenticular degeneration. The results of this study indicated that AFB1 was found to be teratogenic in rabbits when given by oral route during gestation days 6-18 and the dose of 0.1 mg/kg could be considered as the minimum oral teratogenic dose. The histological examination of the fetal tissues indicated its importance in identifying the visceral anomalies which were otherwise not visible.

Isolation of Canine parvovirus with a view to identify the prevalent serotype on the basis of partial sequence analysis.

AIM: The aim of this study was to isolate Canine parvovirus (CPV) from suspected dogs on madin darby canine kidney (MDCK) cell line and its confirmation by polymerase chain reaction (PCR) and nested PCR (NPCR). Further, VP2 gene of the CPV isolates was amplified and sequenced to determine prevailing antigenic type. MATERIALS AND METHODS: A total of 60 rectal swabs were collected from dogs showing signs of gastroenteritis, processed and subjected to isolation in MDCK cell line. The samples showing cytopathic effects (CPE) were confirmed by PCR and NPCR. These samples were subjected to PCR for amplification of VP2 gene of CPV, sequenced and analyzed to study the prevailing antigenic types of CPV. RESULTS: Out of the 60 samples subjected to isolation in MDCK cell line five samples showed CPE in the form of rounding of cells, clumping of cells and finally detachment of the cells. When these samples and the two commercially available vaccines were subjected to PCR for amplification of VP2 gene, a 1710 bp product was amplified. The sequence analysis revealed that the vaccines belonged to the CPV-2 type and the samples were of CPV-2b type. CONCLUSION: It can be concluded from the present study that out of a total of 60 samples 5 samples exhibited CPE as observed in MDCK cell line. Sequence analysis of the VP2 gene among the samples and vaccine strains revealed that samples belonged to CPV-2b type and vaccines belonging to CPV-2.