Efficacy of herbal extracts and closantel against fenbendazole-resistant Haemonchus contortus.

This study assessed the efficacy of closantel vis-à-vis herbal extracts with known anti-parasitic properties, against fenbendazole-resistant nematodes in goats maintained under a semi-intensive system of management at the University goat farm, Jabalpur. Fifty goats were randomly assigned to five groups, each comprising 10 animals, irrespective of their breed, age and sex. Each animal in Group I, II and III was orally administered with aqueous leaf extracts of neem (Azadirachta indica) at 1 g/kg body weight, sitaphal (Annona squamosa) at 1.5 g/kg body weight and tobacco (Nicotiana tabacum) at 1 g/kg body weight, respectively, whereas Group IV was an untreated control group. Each animal in Group V was orally treated with closantel at 10 mg/kg body weight. During the course of the study, all animals were maintained under an identical semi-intensive system of management. Compared to the untreated control group (Group IV), there was no conspicuous reduction in post-treatment (day 10) faecal egg counts (FEC) in animals administered with the herbal extracts (Groups I, II and III), which is suggestive of poor anti-parasitic activity. However, using the faecal egg count reduction test (FECRT), the overall efficacy of closantel was recorded as 95.64%. This supports the rotational use of closantel as a preferred choice over the benzimidazole group of anthelmintics and/or herbal extracts to meet the acute challenge of in situ development of drug-resistant gastrointestinal nematodes, especially Haemonchus contortus.

An assessment of benzimidazole resistance against caprine nematodes in Central India.

Current status of resistance to benzimidazole (BZ) group of anthelmintic drugs against caprine nematodes in Central India at Amanala goat farm, Jabalpur, Madhya Pradesh (M. P.), was systematically investigated using faecal egg count reduction (FECR) test and egg hatch test (EHT). Besides, allele-specific PCR (AS-PCR) was deployed to ascertain the susceptible genotype (alleles) especially of the Haemonchus contortus. Randomly selected 30 goats, irrespective of age and sex, were divided into three groups of 10 each, to serve as treated and untreated controls. It was ensured that the animals were not administered with an anthelmintic drug for the past 3 months prior to undertaking the study, and faecal egg counts were estimated. FECR test evidenced fenbendazole resistance by partial elimination (24.90%) copro-egg counts in the treated group of animals vis-à-vis controls with a lower confidence interval of -26%. Further, EHT revealed ED-50 value of 0.335 µg of thiabendazole/ml, confirming benzimidazole resistance in the animals of that farm. AS-PCR showed that 62% of H. contortus larvae were homozygous resistant (rr), 24% heterozygous (rS) and 14% homozygous susceptible (SS). The genotypic frequencies of three genotypes (rr, rS and SS) were significantly (P < 0.01) different. The prevalence of benzimidazole resistance allele (r) was also significantly (P < 0.01) higher (74%) as compared to susceptible allele (S) (26%). The resistance to benzimidazole has been discussed while emphasizing improved managemental practices designed to reduce exposure of the goat population to parasites, minimize frequency of anthelmintic use at optimum dose and rotational use of different chemical groups of medicines with different mode of action, so as to overcome and combat the upcoming problem in the field.

Reduction of non-enzymatic browning of orange juice and semi-concentrates by removal of reaction substrate.

A study was conducted to standardize the technology for the removal of amino acids (one of the browning reaction substrates) from sweet orange cv. Malta Common juice to reduce colour and quality deterioration in single strength juice and during subsequent concentration. Juice of sweet orange (Citrus sinensis) cv. Malta Common fruits was extracted by screw type juice extractor, preserved in 500 ppm SO2 and clarified by using "Pectinase CCM" enzyme (0.2% for 2 h at 50 ± 2 °C). For removal of amino acids juice was passed under gravity through a glass column packed with an acidic cation exchange resin (CER), Dowex-50 W and quantity to be treated in one lot was standardized. The CER treated and untreated juices were concentrated to 15 and 30°Brix in a rotary vacuum evaporator. Results indicate that 121 ml of orange juice when passed through a glass column (5 cm internal diameter) packed with cation exchange resin (Dowex-50 W) upto a height of 8 cm, could remove about 98.4% of the amino acids with minimum losses in other juice constituents. With cation exchange resin treatment, the non-enzymatic browning and colour deterioration of orange juice semi-concentrates was reduced to about 3 folds in comparison to untreated counterparts. The retention of vitamin C and sugars was also better in semi-concentrates prepared from cation exchange resin treated juice. Thus, cation exchange resin treatment of orange juice prior to concentration and storage is highly beneficial in reduction of non-enzymatic browning, colour deterioration and retention of nutritional, sensory quality of product during preparation and storage.

Intestinal helminths induce haematological changes in dogs from Jabalpur, India.

The effect of canine intestinal helminths on the haematological profile of 200 dogs, of both sexes and variable age, visiting university veterinary clinics for routine examination was investigated. The dogs were assigned to parasitized (n = 39) and non-parasitized (n = 161) groups of animals. Coprological examination revealed a 19.5% prevalence of different species of the helminths. Of these animals, 10.25% had mixed infections with Ancylostoma caninum, Toxascaris spp. and Dipylidium caninum. The intensity of A. caninum infection was the highest, with mean egg counts of 951.43 (standard error 88.66), followed by Toxascaris 283.33 (standard error 116.81) and D. caninum. The parasitized animals had significantly lower levels of haemoglobin, packed cell volume and total erythrocyte counts than non-parasitized animals (P < 0.01). Values of other parameters, except for lymphocytes and eosinophils, were not different between the two groups. Analyses of the haematological profile revealed normocytic hypochromic anaemia in the parasitized group of animals.

Antagonism of Microsporum species by soil fungi.

Eighteen fungi isolated from soil by hair bating method were tested against soil inhabiting Microsporum equinum, Microsporum fulvum, Microsporum gypseum and Microsporum racemosum for their antagonistic interactions. Colony inhibition during dual cultures showed inhibition of all the four Microsporum species. The maximum inhibition of M. equinum, M. fulvum, M. gypseum and M. racemosum was caused by Chrysosporium keratinophilum, Chrysosporium tropicum, Curvularia lunata and Chrysosporium lucknowense in dual cultures. On the other hand, M. fulvum showed maximum inhibition of Macrophomina phaseolina (70.1%) while M. equinum, M. gypseum and M. racemosum showed maximum inhibition of Colletotrichum gloeosporoides. Staling products of C. lucknowense accelerated growth of all Microsporum species, C. keratinophilum 3 and Chrysosporium evolceaunui and M. phaseolina accelerated growth of two species of Microsporum. Staling product of Alternaria alternata was most inhibitory. Culture filtrates of Trichophyton vanbreseughemii accelerated the growth of all the four Microsporum species and C. tropicum, C. lucknowense accelerated growth of two species, while Botryotrichum piluliferum accelerated growth of three species of Microsporum. Volatiles showed inhibition of all the Microsporum species ranging from 0.33 to 57.2% except in case of M. fulvum. Lysis of Microsporum mycelium was the most common feature.

Ni nanoparticle-confined covalent organic polymer directed diaryl-selenides synthesis.

The present work describes the preparation of a new covalent organic polymer (COP) and its application as a hetero support for diaryl selenides synthesis. A nitrogen rich COP (CGP) has been synthesized via SNAr reaction of cyanuric chloride with guanidinium hydrochloride. The successful confinement of COP with Ni nanoparticles through post-synthetic transformations (Ni@CGP) provides excellent catalytic activity for the transformation of aryl halides into diaryl selenides using elemental selenium powder. The synthetic transformations are well confirmed using various modern analytical and spectroscopic techniques which reveal high chemical and thermal durability. The N-rich framework of CGP fortifies the confinement of Ni NPs. Ni@CGP provides an efficient approach for diaryl selenides synthesis using a very cheap selenating reagent under water benign solvent conditions (DMSO : H2O) at room temperature with high reusability. Significantly, our work not only contributes the opportunity for developing economical and effective non-noble metal decorated COPs as heterogeneous catalysts, but also delivers an efficient approach to produce industrially important C-Se coupling products.

Immunodiagnostic/protective role of cathepsin L cysteine proteinases secreted by Fasciola species.

Cathepsin L, a major cysteine proteinase secreted by the parasite plays a pivotal role in various aspects of its pathogenecity. The enzyme takes part in nutrient acquisition by catabolizing host proteins to absorbable peptides, facilitates the migration of the parasite through the host intestine and liver by cleaving interstitial matrix proteins such as fibronectin, laminin and native collagen and is implicated in the inactivation of host immune defenses by cleaving immunoglobulins. Recently, Cathepsin L has been shown to suppress Th1 immune response in infected laboratory animals making them susceptible to concurrent bacterial infections. Accordingly, the protease has been recognized as an important target at which parasite intervention strategies should be directed. Fluke Cathepsin L cysteine proteinases are also reported as sensitive and specific markers for the immunodiagnosis of fasciolosis in ruminants. Further, vaccination of laboratory or large animals with these proteases resulted in a significant reduction in fluke burden and/or fecundity.

In vitro antiplasmodial efficacy of synthetic coumarin-triazole analogs.

Twenty two diverse coumarin-triazole derivatives were synthesized by alkylation of 7-hydroxy-4-methyl-coumarin followed by click chemistry at 7-position. These compounds were evaluated for their in vitro antiplasmodial activity against chloroquine sensitive strain of Plasmodium falciparum (3D7). Compound 9 (7-[1-(2, 4-dimethoxy-phenyl)-1H- [1-3] triazol-4-ylmethoxy]-4-methyl-chromen-2-one) was found most active with IC50 value 0.763 ±â€¯0.0124 µg/mL. Further, the structure of compound 20 was characterized by single crystal X-ray diffraction. In view of impressive results, we considered it worthwhile to validate the results of in vitro antiplasmodial activity by assessing whether these compounds are capable of hampering the catalytic activity of DNA gyrase, thus preventing its supercoiling function.

Preparation of nanosize polyaniline and its utilization for microwave absorber.

Polyaniline powder in nanosize has been synthesized by chemical oxidative route. XRD, FTIR, and TEM were used to characterize the polyaniline powder. Crytallite size was estimated from XRD profile and also ascertained by TEM in the range of 15 to 20 nm. The composite absorbers have been prepared by mixing different ratios of polyaniline into procured polyurethane (PU) binder. The complex permittivity (epsilon' - jepsilon") and complex permeability (mu' - jmu") were measured in X-band (8.2-12.4 GHz) using Agilent network analyzer (model PNA E8364B) and its software module 85071 (version 'E'). Measured values of these parameters were used to determine the reflection loss at different frequencies and sample thicknesses, based on a model of a single layered plane wave absorber backed by a perfect conductor. An optimized polyaniline/PU ratio of 3:1 has given a minimum reflection loss of -30 dB (99.9% power absorption) at the central frequency 10 GHz and the bandwidth (full width at half minimum) of 4.2 GHz over whole X-band (8.2 to 12.4 GHz) in a sample thickness of 3.0 mm. The prepared composites can be fruitfully utilized for suppression of electromagnetic interference (EMI) and reduction of radar signatures (stealth technology).

Acute Intermittent Porphyria in a Kumhar community of Western Rajasthan.

OBJECTIVE: To know the current magnitude of the occurrence of Acute Intermittent Porphyria in Kumhar community of a part of Western Rajasthan. MATERIAL AND METHODS: This is a cross sectional study of rural community of Kumhars of Bikaner district of Western Rajasthan. The households of kumhar community were approached. Besides recording other information and examination details, their urine samples were subjected to Watson-Schwartz Test. RESULT: 1237 subjects out of a total of 2385 kumhar population distributed among 20 randomly selected villages of Bikaner district could be studied. The prevalence of AIP was estimated to be 1.16%. (CI=0.012 +/- 0.0005), showing higher prevalence than the earlier reported studies. The cases exhibited varied symptomatology. Overall preponderance was in favor of females (2:1) and majority of cases (38.9%) were found in the age group of 20-29 years. Average age of manifestation was 24.5 +/- 4.8 years. A follow up was also done to know the disease consequences, if any. CONCLUSIONS: The study finds higher prevalence of the disease and as such warrants a need for generating awareness among the families of cases in the specific caste group in the study area for early detection and better management of the disease.

Campylobacter species associated with diarrhoea in patients from a tertiary care centre of north India.

BACKGROUND & OBJECTIVES: Most laboratories do not routinely distinguish the various Campylobacter species, though almost all Campylobacter species have been isolated from human faeces. The epidemiological and clinical aspects of its infection and the species involved in genesis of diarrhoea are least understood in the developing countries. The aim of the present study was to find out frequency of Campylobacter species isolated from patients with diarrhoea over a 12-year period and to analyse their features. METHODS: Campylobacter strains isolated from stool samples of patients with diarrhoea were identified to the species level on appropriate media at 42 degrees C micro-aerobically. Patients' demography and clinical data were analyzed retrospectively; 25 Campylobacter jejuni strains were tested for toxin production and 23 strains were typed by Penner scheme. RESULTS: A total of 62 strains were isolated from 59 patients and the various species were C. jejuni 51 (82.3%), C. coli 8 (12.9%), C. lari 2 (3.2%), and C. upsaliensis 1 (1.6%). Children < 5 yr of age were most affected (34/59; 57.6%), followed by patients in 15-30 yr of age (12/59; 20.3%). Presentation of watery diarrhoea was significantly more common than inflammatory diarrhoea (50/59, 84.7% vs 9/59, 15.3%; P < 0.001). Recurrence occurred in 3 (5.1%) patients. Majority of the infections resolved within one week; one HIV-positive patient had chronic diarrhoea. Two patients developed Guillain-Barré syndrome following Campylobacter infection. Twenty (80%) of 25 strains were toxigenic and 20 (87%) of 23 strains could be typed by Penner scheme. INTERPRETATION & CONCLUSION: In our patients, 4 different Campylobacter species and various C. jejuni serotypes were involved in gastroenteritis. Majority of the infections were watery diarrhoea and in children < 5 yr of age. There is a need of a population-based systematic study to know the epidemiology of whole spectrum of campylobacters in India.

Entomological studies in a dengue endemic area, Jalore, Rajasthan.

Entomological studies on prevalence of adult and immatures of Aedes aegypti along with associated ecological factors have been conducted in a dengue endemic area of Jalore, Rajasthan from 1992 to 1993. Studies in two areas; reportedly affected and unaffected revealed more adults and higher breeding indices in the affected area as compared to the unaffected one. Mosquitoes harbouring dengue antigen were found only in the affected locality. Presence of dengue antigen in field caught mosquitoes in the affected locality was observed mainly during the months of January to April in both the years. Statistical analysis of data has shown a correlation to be significant between adult house index and breeding index in the affected area while this association was found insignificant in the unaffected locality. Water storage practices of the population due to irregular water supply in the affected area is the possible cause for a higher vector concentration in the locality. Seasonality of occurrence of dengue fever patients in a particular period of the year is associated with a relatively higher vector density and the presence of infected mosquitoes which is due to favourable temperature, relative humidity, water temperature and pH.

Candidaemia in patients of a tertiary health care hospital from north India.

BACKGROUND & OBJECTIVES: Candidaemia is an important cause of mortality in hospital settings. Limited information is available from India on nosocomial candidaemia. The objective of the present study was to isolate and identify yeasts from patients suspected to have nosocomial bloodstream infection (BSI) and to determine the carriage rate of Candida species, risk factors for acquisition of infection and mortality in this group of patients. METHODS: Blood samples from 4871 patients suspected to have BSI at least 48 h after admission were cultured following standard protocol to isolate and identify the pathogens. Clinical details, possible risk factors and outcome of all candidaemic patients were recorded and analysed. Samples of hand washings and throat gargles from these patients were also cultured to determine the carriage rate. Candida albicans isolated from patients and their carriage sites were genotyped by randomly amplified polymorphic DNA (RAPD) analysis to study strain relatedness. RESULTS: Twenty one patients with candidaemia were detected with mortality of 55 per cent. Candidaemia per 1000 admissions was 1.61. Isolation of non-C. albicans Candida species was significantly higher than C. albicans (14/21 vs 7/21: P < 0.05). Use of broad-spectrum antibiotics (43%), gastrointestinal surgery (23%), immunosuppressive therapy (23%), protein calorie malnutrition with parenteral hyperalimentation (23%) and neutropaenia (14%) were identified as probable risk factors. The seven C. albicans strains isolated from patients with BSI were typed into 6 genotypes. Yeast carriage rate among the patients was 71.4 per cent. C. albicans isolated from the hand, throat and blood of two patients had identical genotype. INTERPRETATION & CONCLUSION: BSI due to non-C. albicans Candida species is more common than C. albicans in our patients and candidaemia is associated with high mortality. RAPD appears to be a simple method to study strain relatedness for C. albicans. There is a need for early diagnosis and systematic surveillance to meet the challenges of nosocomial candidaemia.

Role of yeasts as nosocomial pathogens & their susceptibility to fluconazole & amphotericin B.

A total of 213 and 208 yeasts were isolated as nosocomial pathogens from various infected specimens during 1996 and 1997 respectively. Yeasts ranked fifth among uropathogens in both the years and from eighth to eleventh in other specimens. Increasing trend in nosocomial urinary tract yeast infection (11.9 in 1996 to 12.6 in 1997) and decreasing trend in wound and other infections (5.1 in 1996 to 2.9 in 1997) per 1000 patients' discharges were observed; blood stream infection remained unchanged (2/1000 discharges) in both the years. Eighty two (41 from each year) randomly selected yeasts were identified to species level following standard protocol and tested for antifungal susceptibility against fluconazole and amphotericin B by reference broth macrodilution technique and agar dilution (AD) method. The frequency of various yeast species identified was Candida albicans 39 (47.6%), C.tropicalis 29 (35.4%), C. krusei 4 (4.9%), C. glabrata 3 (3.7%), C. zeylanoides 2 (2.4%), C. guilliermondii 2 (2.4%), one strain (1.2%) each of C. kefyr, C. parapsilosis, and Trichosporon beigelii. Resistance to fluconazole (MIC > or = 64 micrograms/ml) as per NCCLS criteria was observed in 2 Candida sp. (2.4%). Significantly higher number of non-albicans Candida sp. (8/43; 18.6%) had MIC > 8 micrograms/ml as compared to C. albicans (2/39; 5.1%) (P < 0.05). Only one strain of C. tropicalis had MIC 8 micrograms/ml to amphotericin B and none had MIC > 8 micrograms/ml. Agreement between the reference and the AD methods for fluconazole was 88 per cent and for amphotericin B was 94 per cent. The present study indicates that Candida sp. are emerging as important nosocomial pathogens and the tendency of yeasts to develop resistance to antifungal agents appears to be a challenge for patient management.

28 kDa Fasciola gigantica cysteine proteinase in the diagnosis of prepatent ovine fasciolosis.

Coprological confirmation of ovine fasciolosis in the field, prior to out breaks of the disease and/or strategic antifluke medication, seem to be of little consequence. Efforts are, therefore, being made to evolve a putative antigen specific to serodiagnostic test for early diagnosis during prepatency. In the present investigation, 28 kDa cysteine proteinase was used in ELI SA and Western blot to detect Fasciola gigantica antibodies and further Dipstick-ELISA was developed for field application, using known positive monospecific sera from experimentally infected sheep with 100 F. gigantica metacercariae. Isolation of 28 kDa cysteine proteinase was achieved from bubalian origin flukes. The specific antigen, recognised homologous antifluke antibodies by Western blot as early as 2nd week post-infection (wpi) with 100% sensitivity, in sera samples of sheep harbouring 38 flukes and by 10th wpi in sheep harbouring 3-8 flukes. All sheep were found positive for the infection when ELISA and/or Dipstick-ELISA was applied from 4th wpi. In pooled sera of infected sheep, these were positive during 4th wpi.

Occurrence of keratinophilic fungi on Indian birds.

Keratinophilic fungi were isolated from feathers of most common Indian birds, viz. domestic chicken (Gallus domesticus), domestic pigeon (Columba livia), house sparrow (Passer domesticus), house crow (Corvus splendens), duck (Anas sp.), rose-ringed parakeet (Psittacula krameri). Out of 87 birds, 58 yielded 4 keratinophilic fungal genera representing 13 fungal species and one sterile mycelium. The isolated fungi were cultured on Sabouraud's dextrose agar at 28 +/- 2 degrees C. Chrysosporium species were isolated on most of the birds. Chrysosporium lucknowense and Chrysosporium tropicum were the most common fungal species associated with these Indian birds. Maximum occurrence of fungi (47%) was recorded on domestic chickens and the least number of keratinophilic fungi was isolated from the domestic pigeon and duck. The average number of fungi per bird was found to be the 0.44.

Cryptococcal infection in patients with clinically diagnosed meningitis in a tertiary care center.

A retrospective analysis of 326 clinically diagnosed cases with meningitis over a period of five-and-a-half years was carried out to determine the prevalence of cryptococcal infection, its associated risk factors and therapeutic outcome. Fifty-four (16.6%) patients with cryptococcal meningitis were identified by smear examination, culture and/or cryptococcal antigen latex agglutination test. Records of 45 cryptococcal meningitis patients were available; 18 (40%) of them were apparently healthy immunocompetent individuals, 13 (28.9%) had human immunodeficiency virus (HIV) infection, 9 (20%) were renal transplant recipients, 4 (8.9%) were diabetic and 1 (2.2%) had systemic lupus erythematosus. Ten (22.2%) patients died and 11 (24.4%) patients (all HIV-positive) left against medical advice. The present study indicates that cryptococcal infection is associated with high mortality. Presenting symptoms being indistinguishable from other causes of central nervous system infection, all patients with a clinical diagnosis of meningitis, irrespective of their immune status should be investigated for cryptococcal infection.

A study of forced vital capacity and its predictors among the sand stone quarry workers.

The study assesses the relationship of selected demographic, anthropometric, radiological, work-history and smoking-history related variables with Forced Vital Capacity (FVC) in a sample of 168 sand stone quarry workers in a desert environment. Except the variables related to smoking behaviour, all are found significantly associated with FVC. The predictors of FVC screened through the multiple regression analysis were age, weight, height, profusion of opacities in chest X-ray and duration of work. Mean FVC was found significantly lower as compared to the healthy adult population. On an average FVC% was 90% of predicted (Percentage deviation of 10%). A clear increasing trend in FVC% along age/duration of work was observed among the young workers, which may be due to building of muscles in this job. Percentage deviation of 20% or more in FVC was significantly associated with presence of radiological opacities suggestive of silicosis (odds Ratio = 3.3). The diagnostic utility of the same is also assessed.

Correlates of peak expiratory flow rate: a study of sand stone quarry workers in desert.

The study assesses the relationship between selected demographic, anthropometeric, radiological, work-history and smoking-history related variables with Peak Expiratory Flow Rate (PEFR) in a sample of 286 sand stone quarry workers from a desert region. Age, height, profusion of radiological opacities, age at beginning of work, duration of work and duration of smoking were found significantly associated with PEFR while duration of exposure to dust, nature of job and number of biries smoked per day failed to establish their significant association. Mean PEFR of these workers was found significantly lower than the same among healthy adults from the study area. The subset of predictors of PEFR only included age, height and profusion of radiological opacities as identified by multiple regression analysis.

Pulmonary echinococcal cyst with a filamentous fungus co-infection.

Fungal infections are known to colonize the pre-existing lung cavities formed as a result of diseases like tuberculosis, sarcoidosis, bronchiectasis and cavitatary neoplasia, mostly encountered in immunocompromised patients. Pulmonary echinococcal cysts have been reported coexistent with cryptococcosis and other saprophytic mycosis, but the coexistence of aspergillosis and echinococcal cyst is extremely rare and occasionally been reported in English literature. Active invasion and proliferation of the fungi in the laminated ectocyst of the echinococcal cyst is very unusual. We report a case of 60 years old immunocompetent female, presented with cough, chest pain and shortness of breath. The chest X-ray showed a large thick walled cavity in the lower and mid zone of right lung with positive water lily sign. Surgical enucleation of the echinococcal cyst revealed aspergilloma involving the cavity with massive invasion of laminated ectocyst by filamentous fungus, morphologically resembling an Aspergillus species and was further treated with Itraconazole for 3 months. This unique coexistence of active pulmonary echinococcosis and aspergillosis is being reported because of its rarity and clinical importance for its management.