Dot-ELISA based on recombinant Hypodermin C of Przhevalskiana silenus for field diagnosis of goat warble fly infestation.

Goat warble fly infestation (GWFI) is an economically important myiasis caused by larvae of Przhevalskiana silenus (Diptera, Oestridae), prevalent in countries of the Mediterranean Basin and Indian subcontinent. GWFI is characterized by the presence of subcutaneous warbles at the lumbar and sacral region of dorsum in the infested animal. The early larval instars (L1 and L2) remain inaccessible to physical detection due to their small size and subcutaneous presence thus causing hindrance in the diagnosis. The objective of present study was to develop a field applicable early diagnostic intervention for GWFI monitoring and prophylactic management for effective control of the disease. Recombinant Hypodermin C (rHyC) antigen of P. silenus was expressed in Escherichia coli. The purified protein was used for optimizing dot-ELISA in a checkerboard titration using goat warble fly infested serum as known positive. The optimized assay was further tested for lower temperature (18°C) and incubation time (30 min). The optimized assay was assessed for inter-rater reliability and field samples. The optimized conditions require 188 ng of protein/dot, 1:800 dilution of serum sample, 1:4000 dilution of anti-goat IgG conjugate and 5% skim milk powder in phosphate buffer saline as blocking buffer. The assay was found to have a diagnostic sensitivity and specificity of 97.3% and 95.8%, respectively. The inter-rater reliability of dot ELISA with rHyC indirect ELISA was found to be almost perfect with a Cohen's kappa index of 0.973. Further testing at ambient temperature (18°C) and shorter incubation steps (30 min) supported suitability of the assay for field diagnosis of GWFI. The present study provides the first report of a sensitive and specific dot-ELISA for early diagnosis of GWFI which is rapid and cost effective. The test may provide an effective field applicable tool for sustainable control of GWFI.

Prevalence and molecular characterization of Eimeria species affecting backyard poultry of Jammu region, North India.

The present study was conducted from January 2018 to December 2019 to know the prevalence of coccidiosis in backyard poultry in Jammu, Samba, and Udhampur districts of Union Territory of Jammu and Kashmir, North India. A total of 600 pooled fecal samples collected from backyard poultry were examined for presence of Eimeria oocysts. Morphometry and Polymerase Chain Reaction (PCR)-based amplification of ITS-1 gene was carried to characterize the Eimeria species infecting the backyard poultry of the study area. An overall prevalence of 28.5% Eimeria spp. infection among backyard poultry birds was recorded. Among the seasons, highest prevalence was recorded during rainy season (32%) with significantly (p < 0.05) high oocyst excretion (1.77 ± 0.01) and lowest during summer (19.3%) with low oocyst excretion (0.17 ± 0.006). Young birds up to 3 months of age were found to be more susceptible to infection than older birds, with a significantly (p < 0.05) high prevalence percentage of 38.02. Morphometry with COCCIMORPH software revealed presence of Eimeria tenella, Eimeria necatrix, Eimeria acervulina, and Eimeria maxima species with prevalence rates of 27.6%, 21.3%, 16.5%, and 3.6%, respectively. The amplified fragments of ITS-1 gene presented different sizes of Eimeria spp. viz. E. acervulina (321 bp), E. tenella (278 bp), E. maxima (145 bp), and E. necatrix (383 bp). The study concluded that although backyard poultry did not show clinical form of coccidiosis, it may act as source of potential reservoir.

First report of Przhevalskiana silenus derived recombinant hypodermin C based indirect ELISA for serodiagnosis of goat warble fly myiasis.

Goat warble fly infestation (GWFI) is a subcutaneous myiasis caused by larvae of Przhevalskiana silenus, an insect belonging to the order Diptera. The diagnosis of GWFI is challenging in the early larval instars (L1 and L2) as they are occult under the skin and hair coat causing prolonged economic loss in form of meat and hide damage. This necessitates early diagnosis for disease control at herd level and its prophylactic management to prevent economic losses. Hypodermins, a class of serine proteases from Hypoderminae subfamily have been used as serodiagnostic antigens for the past four decades for diagnosis of warble fly myiasis. In this study,the immunodominant antigen Hypodermin C (HyC) from P. silenus has been recombinantly expressed in E. coli and immunogenic characterisation of expressed protein was done. The protein shows hallmark residues in conserved cysteine and catalytic triad typical of serine proteases along with similar profile of immunoreactivity towards Hypoderminae infestation. The present study reports an optimised indirect-ELISA based on recombinant HyC derived from P. silenus for early diagnosis of GWFI. The optimised indirect ELISA provides a sensitive and specific immunodiagnostic for mass surveillance of the GWFI with diagnostic specificity and sensitivity of 96% and 100%, respectively and not showing any cross reactivity against other important parasitic and bacterial diseases of goats. This study presents the first report of indirect ELISA based on recombinant Hypodermin C antigen derived from P. silenus for the serosurveillance of goat warble fly disease.

Catalytic Applications of Non-Centrosymmetric Oxide Nanomaterials.

Noble-metal-free catalytic nanoparticles hold the promise being abundant, low-cost materials having a small environmental footprint and excellent performance, albeit inferior to that of noble metal counterparts. Several materials have a long-standing history of success in photocatalysis, in particular titanium dioxide, and in recent years more complex oxides and added functionality have emerged with enhanced performance. We will discuss different approaches related to the use of non-centrosymmetric and polar oxide nanoparticles and how the bulk photovoltaic effect, piezoelectricity, and pyroelectricity add to photocatalysis and tribocatalysis. We pay special attention to discriminate between the role of free versus that of bound charges within the catalyst, which is crucial to disentangle the different contributions to the catalytic reaction for the benefit of the overall enhanced catalytic performance in e.g. wastewater treatment and ultimately water-splitting.

Status of ivermectin resistance in Rhipicephalus (Boophilus) microplus (Acari: Ixodidae) populations from north-western Himalayas, India.

Status of ivermectin resistance was studied in 16 isolates of Rhipicephalus (Boophilus) microplus ticks collected from north-western Himalayan region of India using larval immersion test (LIT). Regression graphs of probit mortality of larval ticks plotted against log values of concentrations of ivermectin were utilized for the determination of slope of mortality, lethal concentrations (LC50 and LC95) and resistance ratios (RRs). Out of 16 isolates examined, 15 were found resistant against ivermectin. Level II resistance was detected in eleven isolates (RR50 ranged from 5.72 to 7.62), whereas four isolates exhibited level I resistance (RR50 ranged from 1.58 to 2.77). Only one isolate was found susceptible (RR50: 1.23). Higher RR values were observed in tick isolates from low altitude areas where intensive animal husbandry activities are practiced as compared to tick isolates of high altitudes accredit the fact that managemental practices, availability of drug and its frequency of application play an important role in development of resistance.

Epidemiology, haematology and molecular characterization of haemoprotozoon and rickettsial organisms causing infections in cattle of Jammu region, North India.

BACKGROUND: The present study was aimed at establishing the prevalence, epidemiology and molecular characterization of major haemoprotozoons (Babesia and Theileria) and rickettsia (Anaplasma) of cattle in Jammu region (North India) using microscopy and Polymerase Chain Reaction (PCR). Hematology, microscopy and PCR based prevalence studies were undertaken with 278 whole blood samples from cattle. Molecular prevalence studies were followed by genetic characterization of the isolates of Babesia, Anaplasma and Theileria spp. based on 18S rRNA, 16S rRNA and Tams1 gene, respectively. The data related to metrology and epidemiological variables like temperature, rainfall, season, age and type of livestock rearing was analyzed and correlated with occurrence of disease by statistical methods. RESULTS: The prevalence based on microscopy was 12.9% (36/278) whereas PCR recorded 30.22% (84/278) animals positive for haemoparasitic infections. All the samples found positive by microscopy were also recorded positive by PCR. Thus the study revealed prevalence of Babesia bigemina, Anaplasma marginale and Theileria annulata to be 9.7, 16.5 and 0.7% respectively. The metrological and epidemiological variables made inroads for the propagation of vector ticks and occurrence of infection. Haematological alterations predominantly related to decrease in haemoglobin, red blood cell count and packed cell volume were evident in diseased animals and collaterally affected the productivity. Further the genetic characterization of Babesia bigemina. (MN566925.1, MN567603, MN566924.1), Anaplasma marginale. (MH733242.1, MN567602.1) and Theileria annulata (MT113479) provided a representative data of the isolates circulating in the region and their proximity with available sequences across the world. CONCLUSIONS: Despite holding much significance to the animal sector, comprehensive disease mapping has yet not been undertaken in several parts of India. The present study provides a blue print of disease mapping, epidemiological correlations and genomic diversity of Babesia bigemina, Anaplasma marginale and Theileria annulata circulating in the region.

Smooth pyroelectric luminescence in LiNbO3 single crystals.

We investigate the phenomenon of pyroelectric luminescence in LiNbO3 single crystals. This faint emission of light due to temperature-induced changes of permanent polarization is induced by different types of charge carrier recombination, outside and inside the crystal. With decreasing atmospheric pressure, the external discharge mechanism transitions from sparse intense gas discharge pulses at ambient pressure, to frequent faint discharges close to 1 mbar, to a continuous emission which is referred to as smooth pyroelectric luminescence. Our experimental setup exposes the crystal to constant positive and negative temperature changes in the range of 360-450 K under high vacuum while simultaneously measuring the surface charge density and the emitted intensity. A microscopic model of the luminescence allows the description of the time-dependent pyroelectric luminescence, in particular the determination of deep trap potentials that are otherwise inaccessible to thermal ionization. Using this model, we show that the behavior of this emission in LiNbO3 crystals is consistent with the release of trapped electrons by the Poole-Frenkel effect from a Dirac-well potential, while the commonly assumed coulombic trap shape is in clear disagreement with both the temporal evolution of the emission as well as the magnitude of the electric field obtained in our measurements.

Alterations in oxidative stress parameters and its associated correlation with clinical disease on experimental Cryptosporidium parvum infection in Swiss albino mice.

The present study was conducted to evaluate the changes in oxidative stress parameters on experimental infection with Cryptosporidium parvum in Swiss albino mice. The mice were divided into four groups viz., group I-IV, each group comprising of 15 mice. Group I mice served as healthy control. In Group II mice, C. parvum oocysts @ 104/os were administered, mice of group III were given dexamethasone @ 30 µg/ml in drinking water whereas group IV mice were given dexamethasone @ 30 µg/ml along with C. parvum oocysts @ 104/os. Significant changes were seen in oxidative stress parameters which included significant increase in LPO and decrease in levels of SOD, CAT and GSH in liver and intestine in group IV mice at 10th DPI when compared to others indicating an important role played by free radical induced oxidative stress in the development of C. parvum infection in mice which was clinically characterized by loss of body condition, profuse bloody diarrhoea and peak oocyst shedding intensity occurring at 10th DPI.

Myiasis of domestic and wild ruminants caused by Hypodermatinae in the Mediterranean and Indian subcontinent.

Myiasis caused by Hypodermatinae flies is an economically important disease affecting domesticated and wild ruminants in countries of the Mediterranean and Indian subcontinent. The adult flies have a short life span of 1 week, and they spread disease by laying their eggs on the coat of animals. Hypoderma spp. primarily lay their eggs on cattle, buffalo, roe deer, red deer and reindeer, while Przhevalskiana spp. lay eggs on the coat of goats. The larvae live as tissue parasites, inducing myiasis in the host, and have a major impact on the productivity and welfare of infested animals. Diagnosis of myiasis is mainly based on clinical examination of the animal and immunodiagnosis using serum and milk samples. Chemical control of this pest is considered the best treatment option. Treatment with microdoses of ivermectin (2µg/kg body weight) combined with immunosurveillance strategies has led to the eradication of bovine hypodermosis in a few countries; however, these flies are still prevalent throughout the northern hemisphere. This paper reviews the current status, diagnosis and control measures for Hypodermatinae myiasis of domestic and wild ruminants in the Indian subcontinent and Mediterranean region.

Synthesis of a NbO Type Homochiral Cu(II) Metal-Organic Framework: Ferroelectric Behavior and Heterogeneous Catalysis of Three-Component Coupling and Pechmann Reactions.

A chiral tetracarboxylic acid ligand, H4L, incorporating the (S)-(+)-2-methylpiperazine moiety in its middle, solvothermally forms a homochiral Cu(II) framework, {[Cu2(L)(H2O)2]·(4DMF)(4H2O)}n (LCu). It forms a non-interpenetrated structure consisting of [Cu2(COO)4] paddle-wheel secondary bonding units (SBUs) with NbO topology. Interestingly, the framework LCu exhibits excellent ferroelectric properties. It shows a remnant polarization (Pr) of ∼3.5 µC cm-2 and a coercive field (Ec) of ∼12 kV cm-1 with a distinct electric hysteresis loop. Dielectric studies of LCu reveal almost frequency-independent behavior with a dielectric constant (εr) of ∼42 and a low dielectric loss (tan δ) of ∼0.04 up to 106 Hz, for potential use in high-frequency applications. In addition, activated framework LCu' having uncoordinated metal sites acts as an efficient heterogeneous catalyst in the three-component coupling of amines, aldehydes, and alkynes, as well as in Pechmann reactions of phenols with ß-ketoesters.

Spin phonon interactions and magnetodielectric effects in multiferroic BiFeO3-PbTiO3.

Here, we report the observation of magneto-dielectric and magneto-structural coupling in (1 - x)BiFeO3-xPbTiO3 i.e.(1 - x)BF-xPT) solid solutions with compositions in the vicinity of morphotropic phase boundary, as manifested by a combination of temperature dependent magnetic, Raman and dielectric measurements. Whilst x-ray diffraction and Raman spectroscopy suggest absence of any structural phase transition between 90-300 K, temperature dependent magnetic studies reveal magnetic anomalies in the solid solutions. These results are complemented by identical observations in the dielectric measurements at similar temperatures indicating a coupling between magnetic and electric order parameters. Further, Raman studies on rhombohedral i.e. x = 0.20 samples reveal a coupling between the magnetic structure and the lattice, causing spin-phonon interactions that are possibly responsible for observed magneto-dielectric effects. Our results illustrate that the phase transitions in BiFeO3-PbTiO3 system are fewer than expected and are attributed to a spatial averaging in an inhomogeneous albeit single-phase material due to clustering of Fe- or Ti-ions on different length scales.

Field efficacy of minidosed pour-on ivermectin and eprinomectin against goat warble fly infestation by Przhevalskiana silenus.

The efficacy of minidose of pour-on ivermectin and eprinomectin formulations against first instar larvae of Przhevalskiana silenus was observed in naturally infested goats in the Jammu region, North India. The study was performed in mid August 2011. A total of 280 goats were randomly divided in to 7 groups of 40 each. Goats of the first three groups were treated with pour-on ivermectin at dosage of 2, 5, and 200 µg/kg body weight, respectively, whereas animals of the fourth to sixth groups were treated with pour-on eprinomectin at 25, 50, and 500 µg/kg body weight, respectively. Group VII animals were kept as untreated control. The results indicated that no warbles were recorded between December 2011 and March 2012 on back of animals treated with pour-on preparations of ivermectin at dosage of 5 and 200 µg/kg body weight, respectively, and eprinomectin at dosage of 50 and 500 µg/kg body weight, respectively. Thus, it is concluded that administration of minidose of pour-on ivermectin (5 µg/kg body weight) and eprinomectin (50 µg/kg body weight) is cost effective and so can be used for warble fly control campaign in Jammu region.

Microstructure and interfacial chemistry of pure and La-doped BiFeO3 thin films.

We report on the microstructure and interfacial chemistry of thin films of pure and La-doped multiferroic bismuth ferrite (Bi1-x Lax FeO3 or BLFO), synthesized on Indium Tin Oxide-coated glass substrates by solution-deposition technique and studied using scanning transmission electron microscopy. Our results show that undoped and La-doped thin films are polycrystalline with distorted rhombohedral structure without any presence of any line or planar defect in the films. In addition, the films with La doping did not show any structural change and maintain the equilibrium structure. Cross section compositional analysis using X-ray energy dispersive spectrometry did not reveal either any interdiffusion of chemical species or formation of reaction product at the film-substrate interface. However, a closer examination of the microstructure of the films shows tiny pores along with the presence of approximately 2-3 nm thin amorphous layers, which may have significant influence on the functional properties of such films.

Prevalence of Hypoderma lineatum in cattle of Jammu region.

Clinicoparasitological examination of 750 cattle from Jammu region revealed prevalence of Hypoderma lineatum infestation in 9.73 %. Cattle from Jammu (plain irrigated area) district (397) did not reveal any infestation of hypodermosis, but in Samba district (semi hilly arid area) 20.63 % (73/353) animals harbored H. lineatum larvae. The warbles were recorded from October to March and the mean number of warbles per animal was 7.07 ± 0.45. Infestation significantly varied in different age groups and free grazing practice of animals was one of the risk factor for hypodermosis in cattle of Jammu region.

Economic impact of Przhevalskiana silenus infestation in native goats of Northern India.

Economic losses incurred by Przhevalskiana silenus in goats of Jammu province of Jammu and Kashmir (North India) were assessed from July 2005 to June 2006. Abattoir studies revealed holes in hides for 112 days (mid-December 2005 to first week of April 2006) and trimming of carcass for 243 days (July 2005 to February 2006) during a lifecycle, leading to annual losses of Rs. 7,391,432 and Rs. 10,510,837, respectively. An effect on production traits (weight gain and milk) was assessed on naturally P. silenus-infested Beetal goats. The mean body weight loss of warbled goats was 2.40 kg (equal to 1.096 kg of meat, assuming 45.7% dressing percentage of goat) leading to an annual loss of Rs. 35,473,265. The mean milk yield loss per warbled goat per day was 101.7 g resulting in losses to the tune of Rs. 19,879,012. It can be concluded that P. silenus infestation resulted in an overall loss of Rs. 73,254,547, i.e. 73.26 million annually (equivalent to US $1.62 million; US $1 = Rs. 45.24), where reduced growth alone contributed 49% of the total losses. A study recommends that a control scheme against P. silenus infestation in goats using an effective insecticide should be followed to prevent huge economic losses in hide, milk and meat production. Economic losses associated with P. silenus infestation in goats were estimated for the first time from India.

Prevalence and biology of goat warble fly infestation by Przhevalskiana silenus in Jammu province, India.

Examination of 3,960 goats brought from six districts of Jammu province and slaughtered at Jammu abattoir from July 2005 to June 2006 revealed 47.22% prevalence of larval stages of warble fly (Przhevalskiana silenus Brauer, 1858) in goats. Highest prevalence was recorded in Udhampur (56.44%) and lowest in Doda district (13.49%). Classification of the study area into four zones based on height (meters above sea level), viz., zone I (300-325 m asl), zone II (325-800 m asl), zone III (800-1,500 m asl), and zone IV (>1,500 m asl), revealed significant (p < 0.01) difference in infestation among animals of zone II (71.68%), zone III (40.12%), and zone IV (22.41%). However, animals of zone I did not reveal any infestation. Statistical analysis in relation to age showed significant (p < 0.01) difference among different age groups, i.e., <1 year (2.81%), 1-3 years (51.17%), and >3 years (43.16%). Breed-wise analysis also showed significantly (p < 0.01) higher infestation rate among Bakerwali (51.51%) goats as compared to Beetal (42.59%). But no significant difference was recorded among male (47.81%) and female (46.82%) animals. The overall mean larva count (L1, L2, and L3) was observed to be 14.72 ± 0.34, ranging from 4 to 72. It was significantly (p < 0.05) higher in animals of zone II (22.20 ± 0.21) as compared to goats of zones III (14.21 ± 0.41) and IV (7.73 ± 0.90). Age-wise analysis of mean larva count also showed significant (P < 0.05) difference between animals of 1-3 years (16.25 ± 0.37) and >3 years of age (13.18 ± 0.40). Mean larvae count in relation to sex and breed did not reveal any significant difference. First-instar larvae (L(1)) were recorded from May to mid-September, second larval stage (L(2)) from mid-September to mid-December, and third-stage larvae (L(3)) from mid-December to February. However, No larvae were recorded from March to April, which is suggestive of pupation period in this region. Thus, it is concluded that adult fly is active in April to June. The results further confirmed that internal life cycle of P. silenus is subcutaneous and no migration of larvae occurs. This is the first report from India, based on slaughter house study on the prevalence and biology of goat warble fly infestation. It will help in devising suitable prophylactic and eradication program to check the economic losses rendered by adult fly.