Enhancement of Single Molecule Raman Scattering using Sprouted Potato Shaped Bimetallic Nanoparticles.

Herein, for the first time, we report the single molecule surface enhanced resonance Raman scattering (SERRS) and surface enhanced Raman scattering (SERS) spectra with high signal to noise ratio (S/N) using plasmon-active substrates fabricated by sprouted potato shaped Au-Ag bimetallic nanoparticles, prepared using a new one-step synthesis method. This particular shape of the nanoparticles has been obtained by fixing the amount of Au and carefully adjusting the amount of Ag. These nanoparticles have been characterized using scanning electron microscopy, extinction spectroscopy, and glancing angle X-ray diffraction. The single molecule sensitivity of SERS substrates has been tested with two different molecular Raman probes. The origin of the electromagnetic enhancement of single molecule Raman scattering in the presence of sprouted shape nanoparticles has been explained using quasi-static theory as well as finite element method (FEM) simulations. Moreover, the role of (i) methods for binding Raman probe molecules to the substrate, (ii) concentration of molecules, and (iii) Au-Ag ratio on the spectra of molecules has been studied in detail.

Identification of a novel cluster of PCV2 isolates from Meghalaya, India indicates possible recombination along with changes in capsid protein.

Documentation of the emergence of Porcine circovirus 2 (PCV2) infection and economic losses incurred due to high mortality has been reported worldwide. The prevalence and genetic diversity of the virus has been reported in Northeast India including the possible chances of Classical swine fever virus (CSFV) vaccine failure in pig population in this region resulting in major disease outbreak. Irrespective of the genetic variability, the emergence of a novel cluster (based on the ORF2 phylogeny) was reported last year. The present study describes a state-wide (Meghalaya, India) molecular epidemiological investigation of PCV2 strains in pig population by amplification, sequencing and undertaking phylogenetic analyses. The results indicate the identification of a novel cluster of PCV2 originating from the inter-genotypic recombination between PCV2c and PCV2d. Multiple sequence alignment of amino acids indicates possible substitution in the A, B and C domains of the capsid protein. Molecular structural modelling of the capsid protein of PCV2 indicated possible motif variations in the secondary structure including presence of a tunnel, encountered at the interface region on each chain facilitating in transportation of molecules and acting as an active site for attachment and penetration. The baseline data strengthens the existing control programme of PCV2 and is possibly helpful in the planning of active surveillance strategy in this region.

Detection of classical swine fever virus E2 gene in cattle serum samples from cattle herds of Meghalaya.

The present study focused on the detection and genetic characterisation of 5' untranslated region (5'UTR) and E2 gene of classical swine fever virus (CSFV, family Flaviviridae, genus Pestivirus) from bovine population of the northeastern region of India. A total of 134 cattle serum samples were collected from organised cattle farms and were screened for CSFV antigen with a commercial antigen capture enzyme linked immunosorbent assay (Ag-ELISA) and reverse transcription-polymerase chain reaction (RT-PCR). A total of 10 samples were positive for CSFV antigen by ELISA, while all of them were positive in PCR for 5'UTR region. Full length E2 region of CSFV were successfully amplified from two positive samples and used for subsequent phylogenetic analysis and determination of protein 3D structure which showed similarity with reported CSFV isolate from Assam of sub-genogroup 2.1, with minor variations in protein structure.

Gastrointestinal parasitism of goats in hilly region of Meghalaya, India.

AIM: The aim of this study was to determine the prevalence of gastrointestinal (GI) parasitic infections in goats of hilly region of Meghalaya. MATERIALS AND METHODS: A total of 834 fecal samples of goats were screened for 1 year (2014-2015) using flotation techniques. RESULTS: The overall prevalence of GI parasitic infections in goats was 28.65%. Season-wise highest infections were recorded during rainy season (34.92%) followed by cool (26.87%), hot (26.62%), and cold (20.39%) seasons. Helminths and protozoa infections were recorded in 63.60% and 23.02% animals, respectively. Among the helminths, Strongyle spp. (32.63%) was recorded highest followed by Trichuris spp. (12.55%), Moniezia spp. (10.04%), and Trichuris spp. (8.36%). Among protozoa, only Eimeria spp. was detected. Seven different species of Eimeria spp. were identified, viz., Eimeria christenseni, Eimeria hirci, Eimeria caprina, Eimeria jolchijevi, Eimeria ninakohlyakimovae, Eimeria arloingi, and Eimeria kocharii for the first time from Meghalaya. Maximum egg per gram and oocyst per gram of feces were recorded in the month of August (932.4) and September (674.05), respectively. Mixed infections were recorded in 13.38% samples. Coproculture of goat fecal samples revealed the presence of Haemonchus contortus (72.16%), Oesophagostomum spp. (14.41%), Strongyloides spp. (8.91%), and Trichostrongylus spp. (4.50%) larvae. CONCLUSION: This study indicates that GI helminths and protozoa infections are prevalent in goats of this hilly region of Meghalaya, throughout the year and highly prevalent during rainy season.

On the recovery of horsehair worms, Gordius sp. (Nematomorpha: Gordiida) from pork in Shillong, India.

The members of the phylum Nematomorpha, called as horsehair or gordian worms, are the parasites of arthropods and emerge from their host for reproduction and early development in water. There are about 360 species of nematomorphs described till date. Although, horsehair worms are parasites of arthropod species, sporadic cases of their occurrence have also been reported from several other hosts, including man. This paper describes a case history of the occurrence of two nematomorph worms in the meat of a pig in Shillong, India. The worms were reported to have emerged in live condition from pork bought by a consumer from local market in Shillong. One of the recovered specimens was studied by scanning electron microscopy for species determination and identified as Gordius sp., mainly on the basis of flat polygonal areoles and fine bristles on the cuticle.

Sarcoptic mange infestation in pigs: an overview.

Sarcoptic mange infestation in pigs is caused by Sarcoptes scabiei var. suis. It is the most common mange infestation of pigs. The parasite is distributed worldwide. Pig owners are generally concerned about the internal parasitic infections and ignored the external parasitic infestations. But the external parasitic infestation with S. scabiei var. suis has economic significance as it causes morbidity, mortality, decreased fertility and feed conversion ratio in pigs. Keeping in view of importance of S. scabies var. suis infestation in pigs, this communication discussed about the present and past research works done on S. scabies var. suis infestation in pigs, particularly its prevalence, life cycle, pathological lesions, clinical symptoms, haematobiochemical changes, diagnosis, treatment and control, to have an idea about this infestation at a glance. It has been concluded that the research work done on sarcoptic mange infestation in pigs in India is less in comparison to other countries. It may be due to its consideration as a neglected parasite or due to it's under report. Organization of awareness programs for the farmers by extension personalities or other authorities might be able to save the farmers from economic losses due to this infestation.

Complete genome sequence of emerging porcine circovirus types 2a and 2b from India.

We report here the first characterized complete genome sequence of porcine circovirus types 2a and 2b from northeastern states of India. These isolates may serve as a potential reference for the Indian strains of porcine circovirus types 2a and 2b.

Babesia infection in naturally exposed pet dogs from a north-eastern state (Assam) of India: detection by microscopy and polymerase chain reaction.

The objective of the study was to detect Babesia infections in pet dogs of a north-eastern state of India. The diagnostic efficacy of Babesia infection by polymerase chain reaction (PCR) technique has been compared with microscopy examination. For this, a total of 111 blood samples of pet dogs presented at clinical complex of the College of Veterinary Science, Guwahati, Assam with clinical signs suspected for Babesia infection subjected to the study. A total of 44 (39.63 %) dogs were diagnosed as positive for Babesia infections after microscopic examination. Among these, Babesia canis infection was diagnosed in 5 dogs (4.50 %) and B. gibsoni infection in 39 (35.13 %) dogs microscopically in Giemsa stained blood smears. Molecular diagnosis using PCR detected 63 (56.75 %) dogs positive for Babesia infection. Single infection with B. canis was found in 9 (8.10 %) dogs while B. gibsoni alone was detected in 3 (2.70 %) dogs. Mixed infections by both these species were detected in 51 (45.94 %) dogs. Overall, PCR detected 54 (48.64 %) dogs as B. gibsoni and 60 (54.05 %) dogs as B. canis positive.

Human consumption of rumen flukes of cattle in India.

The practice of eating rumen flukes of cattle by a section of people living in Meghalaya, a north eastern State of India, is reported in this communication. Economically backward, some rural people belonging to Khasi, Jaintia, Garo, and Karbi tribes of Christian and Nepali communities who eat beef are accustomed to consuming cooked flukes during breakfast, meals, and also along with rice beer or alcohol. Inspection of the rumens of cattle during slaughter indicated a prevalence of flukes belonging to Cotylophoron, Paramphistomum, Calicophoron, Gastrothylax, and Fischoederius genera in 74% cases, and their collection from rumen ranged approximately from 50 g to 600 g. Biochemical analysis of flukes found 12.60% total protein, 0.78% fat, and 0.87% ash on fresh weight basis. High prevalence of flukes, easy visualization in rumen, their bulk collection, presence of nutritive value, absence of any ill effect, and lack of imminent danger of transmissibility are believed to be the rationales influencing their consumption by people. It is suggested that dietary benefits obtained from flukes might contribute to the energy transfer and inclusion in the food web.

Sarcoptic mange infestation in pigs in a hilly region of Meghalaya.

Skin lesions were collected from 15 adult crossbred pigs maintained at the pig farm of ICAR Research Complex for the North-Eastern Hill Region, Umiam, a hilly region of the state of Meghalaya, India. Skin scrapings of these 15 animals were examined by KOH digestion method. Presence of mites were detected in five animals which were indistinguishable from Sarcoptes scabiei var. suis. Infected pigs were treated with a single dose of Ivermectin at 200 ug/kg body weight, subcutaneously. No mites were recovered from treated pigs after a period of 10 days of post-treatment. The presence of S. scabiei var. suis in pigs from this part of the country has never been reported. This infestation has great significance because it affects the growth rate and feed conversion significantly, specially to the grower and weaned pigs. It can be concluded that S. scabiei var. suis infestation is prevalent in hilly region of Meghalaya. The pig producers are generally unaware about the severity of the problem of mite infestation, so precaution should be taken to prevent their valuable pigs from S. scabiei var. suis infestation.

Detection of Trypanosoma evansi infection in clinically ill cattle, buffaloes and horses using various diagnostic tests.

This study was undertaken to detect Trypanosoma evansi infection in clinically ill cattle, buffaloes and horses that were suspected of 'surra' infection. Various diagnostic tests were utilized in order to assess the prevalence of infection in the clinically ill and 'suspected' animals. Animals in the Eastern Region of India were systematically studied for T. evansi infection, which had not been tested for previously. Examination of Giemsa-stained blood smears detected 5.3%, 9.4% and 40.6% infections; the mouse inoculation test detected 18.4%, 15.6% and 46.9% infections whereas an in-house ELISA detected anti-trypanosomal antibodies in 42.1%, 43.8% and 65.6% infections in clinically ill buffaloes, cattle and horses, respectively. It can be concluded that T. evansi is prevalent in cattle, buffaloes and horses in the Eastern Region of India.

Endemic status of Trypanosoma evansi infection in a horse stable of eastern region of India--a field investigation.

Diagnosis of Trypanosoma evansi infection in a horse stable of Eastern Region of India on the basis of examination of Giemsa stained blood smears have been done. A high percentage (12.74%) of horses of this stable was found suffering from T evansi infection. This high prevalence of T evansi in horses, in this area could be considered as an alarming situation which has never been explored previously in horses of Eastern Region of India. After a period of 2 months and 18 days of treatment with quinapyramine sulphate and quinapyramine chloride, reinfection with T evansi in treated horses of this stable were noticed. Clinical signs of affected horses and possible causes of reinfection have been discussed.

Larval migration of Toxocara canis in piglets and transfer of larvae from infected porcine tissue to mice.

Visceral larva migrans (VLM), caused by Toxocara canis larvae in humans, animals and birds, is now well documented throughout the world. Seven piglets were infected orally with 5 x 104 embryonated eggs and the migration and distribution of T. canis larvae in the tissues were evaluated. After artificial gastric juice digestion, larval yields at necropsy from different organs and muscles on days 1, 3, 7, 15 and 30 post-infection (DPI) revealed 3.05, 0.97, 0.21, 0.13, 0.05, 0.14% recovery from liver, lungs, heart, kidneys, skeletal muscles and brain tissues respectively, with a total of 2486 (4.97%) recovery from all tissues together. The highest number of larvae 1527 (3.05%) was recovered from the liver throughout the period (1-30 DPI), indicating a special affinity of larvae for the liver. Subsequently five mice were each infected orally with 5 g of infected pig liver and, after necropsy on 10 DPI, 20 +/- 3.62, 17 +/- 5.10, 3 +/- 1.26, 12 +/- 3.92 and 30 +/- 5.69 larvae were recovered from liver, lungs, heart, brain and muscles, respectively. Thus, primarily, the migratory potential and adaptation of T. canis larvae in porcine tissue was examined and, subsequently, their establishment in the second paratenic host, the mouse, has been successful. No influence of host sex on the migratory potential of T. canis larvae was observed. The related pathology caused by migratory larvae and its zoonotic significance through the consumption of raw or undercooked pork has been emphasized.

Characterization of immunogenic proteins of Trypanosoma evansi isolated from three different Indian hosts using hyperimmune sera and immune sera.

The western blot analysis for identification of immunogenic proteins in whole cell lysate (WCL) antigens (Ags) prepared from the Trypanosoma evansi of buffalo, horse and cattle origins using hyperimmune sera (HIS) showed 11 immunogenic proteins and naturally T. evansi infected immune sera (IS) of horse detected 19 immunogenic proteins. HIS and IS of horse recognized five common immunogenic proteins of relative molecular weight (M(r)) ranges 61-64, 44-47, 33-34, 25-26 and 14-16 kilo Dalton (kDa). HIS rose against WCL Ags of T. evansi of buffalo origin and immune sera of horse cross reacted with WCL Ags of T. evansi of different host origin. It can be concluded that in comparison to HIS, IS of horse could able to detect more numbers of immunogenic proteins and five common immunogenic proteins in WCL Ages of T. evansi of different hosts origin. The evidence of higher reactivity of IS in comparison to HIS against T. evansi is being reported for the first time.

Experimental infection of the cockroach Periplaneta americana with Toxocara canis and the establishment of patent infections in pups.

The possible role of the cockroach Periplaneta americana in the transmission of Toxocara canis eggs and larvae via faeces and tissue migration was studied. Cockroaches fed with 3 x 105 and 5 x 105 embryonated eggs were found to harbour viable eggs and larvae from days 1 to 5 post-infection (DPI). At necropsy on 5 DPI, eggs and larvae were also recovered from the rectal contents but not from the tissues of cockroaches. In addition patent infections were established in pups fed on infected faeces of cockroaches, with eggs first appearing in the faeces of pups at 38 DPI. Adult worms of T. canis were also recovered at necropsy. Therefore the importance of cockroaches as good mechanical disseminators of ascarid eggs, especially T. canis, is discussed.

Haemobilia due to a large gall bladder polyp.

A 61-year-old male presented with recurrent bouts of haematemesis and melaena for last 10 days. On examination, he was found anaemic and had jaundice and a mass felt in the right hypochondrium. He required multiple blood transfusions. Oesophagogastroduodenoscopic (OGD) evaluation demonstrated no lesion, no fresh bleeding noticed from ampulla of Vater. Ultrasonographic evaluation demonstrated a heterogeneous mass within the gall bladder lumen with a chink of free lumen between the mass and gall bladder wall and also dilated biliary tract. Laparotomy confirmed mass within the gall bladder lumen without any lymph node involvement at porta or any liver metastasis, and had dilated common bile duct. Cholecystectomy was carried out. The cut section of the gall bladder showed a polypoid mass (5.5 cm x 4 cm) projecting inside lumen having a haemorrhagic spot on irregular surface and which was attached with fundus of the gall bladder through a small stalk. Choledochotomy showed only blood clots within the duct-choledochoduodenostomy was also done. Histopathological examination showed adenomatous polyp with foci of carcinoma in-situ or ly without any evidence of local metastasis. Follow-up was uneventful till after 11 months.

Argyrophilic nucleolar organiser regions (AgNORs) in breast lesions.

Nucleolar organiser regions (NORs) demonstrated by argyrophilia of NOR-proteins are indicator of cellular proliferative activity. The NORs can be identified in the nuclei as brown or black dots with silver colloidal staining technique in formalin fixed paraffin sections and in cytology smears. Seventy-five cases including 45 tissue sections and 30 fine needle aspiration cytology (FNAC) smears of benign and malignant lesions of breast have been studied to evaluate the significance and practical application of AgNOR count per nucleus. Out of 45 tissue sections 15 belonged to fibrocystic disease, 10 fibro-adenomas and 20 carcinomas and of the 30 FNAC smears, 10 were fibrocystic disease, 8 fibro-adenomas and 12 carcinomas. In fibrocystic disease the mean AgNOR count was 1.60 (FNAC group-0.75, tissue section-1.61). In fibro-adenomas it was 1.61 (FNAC-1.63, tissue section-1.59). The mean count in carcinoma was 12.10 (FNAC-12.08, tissue section-12.10). The difference in AgNOR count in fibrocystic disease and fibro-adenoma was not significant, but that between benign breast lesion and carcinoma was significant. No difference was observed between FNAC and tissue section groups in benign or malignant lesions. The simple staining technique can be used as an additional criterion to differentiate the benign and malignant lesions of breast.

PIP: Nucleolar organizer regions (NORs) are loops of DNA encoding ribosomal RNA genes located in the short arms of 5 acrocentric human chromosomes. NORs demonstrated by the argyrophilia of NOR-proteins are indicative of cellular proliferative activity. The NORs can be identified in nuclei as brown or black dots with silver colloidal staining technique in formalin fixed paraffin sections and in cytology smears. 75 cases, including 45 tissue sections and 30 fine needle aspiration cytology (FNAC) smears of benign and malignant breast lesions, were studied to evaluate the significance and practical application of AgNOR count per nucleus. 15 of the 45 tissue sections were of fibrocystic disease, 10 were fibroadenomas, and 20 were carcinomas. Of the 30 FNAC smears, 10 were fibrocystic disease, 8 were fibroadenomas, and 12 were carcinomas. The mean AgNOR count in fibrocystic disease was 1.60; 1.61 in fibroadenomas and 12.10 in carcinomas. Although the difference in AgNOR count between fibrocystic disease and fibroadenomas was insignificant, that between benign breast lesion and carcinoma was significant. No significant difference was observed between FNAC and tissue section groups in benign or malignant lesions. This simple staining technique can be used as an additional criterion to differentiate the benign and malignant lesions of breast.