Haematology, blood biochemistry and tissue histopathology of lambs maintained on diets containing an insect controlling protein (Cry1Ac) in Bt-cottonseed.

This experiment assessed the effect of feeding genetically modified cottonseed (Bt) containing an insect controlling protein (Cry1Ac) on haematology, blood biochemistry and histopathology of lambs. Haemato-biochemicals were estimated at periodic intervals, and histopathology at termination of experiment. Thirty three weaner lambs were fed a composite feed mixture (CFM) ad libitum individually, in three groups for 123 days. The isonitrogenous CFM had roughage (Perl Millet Stover) and concentrate ratio of 350:650. Diet fed to control lambs contained groundnut oil meal as protein source while other two groups received diet containing either whole cottonseed (N-Bt) or Bt-cottonseed (Bt-CS). Daily feed intake and average daily gain were similar among lambs of three groups. Lambs fed N-Bt diet had higher (p < 0.05) serum protein and globulin compared to control and Bt diets, while albumin content was higher (p = 0.018) in Bt diet fed lambs. Serum urea and creatinin content, alkaline phosphatase (ALP) and serum glutamate pyruvic transaminase (SGPT) activities were not different among lamb groups, while urea and creatinin content and ALP activities increased linearly (p < 0.001) with increased feeding period. Blood haemoglobin (Hb), haematocrit (Hc), white blood cells (WBC), mean corpuscular volume (MCV) and mean corpuscular haemoglobin concentration (MCHC) ranged from 11.1% to 11.2%, 31.8% to 32.8%, 7.0 to 8.3 (× 10(3) /µl), 19.1 to 22.5 fl and 33.2% to 35.5%, respectively, were similar among lamb groups. Erythrocyte sedimentation rate (ESR) (p = 0.008) and red blood cell counts (p = 0.006) were higher in Bt diet fed lambs. Control and N-Bt diet fed lambs had mild fatty infiltration in liver and/or micro-calculi in renal cortex, and such lesions were not seen in Bt diet fed lambs. Growth, haemato-biochemical and histopathology did not change by Bt-CS feeding in growing lamb. However, before recommending the use of Bt-CS in routine feed formulations prolonged feeding experiments of Bt-cotton seed require.

Influenza A H1N1 virus in Indian pigs & its genetic relatedness with pandemic human influenza A 2009 H1N1.

BACKGROUND & OBJECTIVES: With the emergence of a new reassortant influenza A H1N1 virus that caused the 2009 pandemic it was felt necessary that pigs should be closely monitored for early detection of any influenza virus infection. Therefore, we investigated disease outbreaks with clinical history suggestive for swine influenza reported to our laboratory by owners of affected pig farms in Uttar Pradesh. METHODS: Detection of swine influenza A virus (SIV) was attempted by isolation in embryonated chicken eggs. Presence of virus was detected by haemagglutination (HA) test and RT-PCR for amplification of different gene segments, cloning and sequencing. BLAST analysis of sequence data, phylogenetic analysis and mutation analysis based on HA, NA and matrix genes was done. RESULTS: SIV could be isolated from one farm and all eight gene segments amplified by RT-PCR. BLAST analysis of partial nucleotide sequences and phylogenetic analysis using nucleotide sequence of HA (601 nt), NA (671 nt) and M (1031 nt) genes indicated close genetic relationship of the Indian swine isolate (A/Sw/UP-India-IVRI01/2009) with human pandemic 2009 (H1N1). The HA gene showed close relationship with the viruses of "North American Swine" lineage, whereas the NA and M genes clustered with the viruses of "Eurasian Swine" lineage, indicating a novel HA-NA reassortant. The remaining of 5 genes (NP, PA, PB1, PB2 and NS) belonged to "North American Swine" lineage. INTERPRETATION & CONCLUSIONS: This is perhaps the first report describing swine influenza among Indian pigs caused by an influenza A H1N1 virus sharing close homology with the human pandemic (H1N1) 2009 virus. Further reassortment with circulating influenza viruses must be closely monitored.

Pathological studies on Bovine papilloma virus-fern interaction in hamsters.

Early pathological changes of Bovine papilloma virus (BPV-2)-fern (Pteridium aquilinum and Onychium contiguum fern) interaction in hamsters were studied. In bracken-exposed cattle, BPV induces malignancy in gastrointestinal and urinary bladder mucosa. Cutaneous warts were transmitted successfully in hamsters approximately after 3 months post inoculation while urinary bladder tumour of enzootic bovine haematuria cases were not transmitted. Histologically, tumour was diagnosed as fibroma. Onychium produced more pronounced effects than bracken fern which was characterized by significant reduction in body weight and testicular atrophy. BPV-fern interaction was not appreciable during early period of tumour induction and requires long-term studies for 12 to 18 months.

Sleep deprivation induces changes in 5-HT actions and 5-HT1A receptor expression in the rat hippocampus.

A 12h sleep deprivation enhances the expression of 5-hydroxytryptamine (5-HT) 1A receptors in rat hippocampus that recedes with 48h sleep recovery. The depressant effect of applied 5-HT on the field excitatory postsynaptic potentials recorded in the CA1 area, is also enhanced in hippocampi of SD rats. Following a 24 or 48h sleep recovery, the increase in the 5-HT effect subsided. These results have implications for therapeutics treating clinical depression.

Proliferative urocystica and adenoma in a guinea-pig.

Proliferative urocystica and adenoma occurred in the urothelium of one clinically normal guinea-pig in a group of nine fed with a diet containing shade-dried Christella dentata (Forssk) fern. It was concluded that the lesions were due to prolonged feeding (285 days) of the fern.

Detection of bovine papilloma viruses in wart-like lesions of upper gastrointestinal tract of cattle and buffaloes.

In present investigation, etiopathological characterization of upper gastrointestinal tract (GIT) tumours of cattle and buffaloes was undertaken. A total of 27 GIT wart-like lesions in rumen, reticulum, mouth and oesophagus of cattle and buffaloes revealed the presence of small nodular to larger spherical or slender growths with thin base present on mucosa and ruminal pillar. Histopathologically, these cases were diagnosed as fibropapilloma/papilloma. This is the first world record on ruminal papillomatosis in buffaloes. Ruminal warts of cattle and buffaloes revealed the presence of BPV-5, -1 & -2, which is the first report of presence of these BPVs in the ruminal warts from India. Quantitative real-time PCR revealed that DNA samples of different GIT wart-like lesions contained varying amount of BPV DNA copy numbers. Immunohistochemistry revealed that the PCNA and Ki67 immunopositivity was present in the basal and spinosum layer of the fibropapilloma/papilloma, indicating these as the cellular proliferation site. In conclusion, the present investigation revealed that BPV-5, -1 & -2 are associated with certain ruminal wart-like lesions/growths in cattle and buffaloes, and the basal and spinosum layer of the ruminal fibropapilloma/papilloma were cellular proliferation sites.

Genome sequence of classical Swine Fever virus genotype 1.1 with a genetic marker of attenuation detected in a continuous porcine cell line.

The complete genome sequencing and analysis of a classical swine fever virus (CSFV) detected in a porcine kidney cell line revealed a close relationship with genotype 1.1 viruses circulating in India and China. The presence of consecutive T insertions in the 3' untranslated region (UTR), as seen in vaccine strains of CSFV, suggested some degree of attenuation.

Genetic characterisation and phylogenetic analysis of PCV2 isolates from India: indications for emergence of natural inter-genotypic recombinants.

Porcine circovirus type 2 (PCV2), the necessary agent in pathogenesis of porcine circovirus diseases (PCVDs), has a worldwide distribution and is considered as one of the most important emerging viral pathogens of economic importance. PCV2 has been divided into four major genotypes namely PCV2a with five clusters or subtypes (2A-2E), PCV2b with three clusters (1A-1C), PCV2c and PCV2d, based on capsid (cap) gene analysis. PCV2 genome is rapidly evolving through events of recombination and mutation. Though, PCV2a was the predominant genotype initially, PCV2b shared majority of PCV2 sequences submitted to GenBank since 2003. In India, data regarding molecular characterisation of PCV2 is scant or absent. In the present study, we thoroughly analysed genetic heterogeneity of PCV2 strains circulating in Indian pig population. The results revealed that pigs in this region harboured PCV2 viruses of different genotypes including PCV2a-2D, PCV2b-1C and PCV2d. More interestingly, two isolates (PCV2Izn-89-13 and PCV2Izn-218-13) were classified as recombinant strains. Further detailed analysis suggested that these strains evolved from inter-genotypic recombination between PCV2a-2C and PCV2b-1C genotypes within cap gene. This study reports for the first time, the emergence of recombinant PCV2 strains in the Indian pig population.

Polycystic liver disease in golden hamsters.

Seven cases of multiple hepatic cysts of varying sizes and shapes were recorded in a closed colony of golden hamsters. Ante-mortem examination failed to show any clinical signs except for abdominal enlargement in one hamster. Multiple, thin-walled cysts of varying sizes (0.25 to 3.0 cm) and shape were observed in the liver. The larger cysts protruded from the liver surfaces and contained a colourless, clear, serous fluid and caused pathological change in 5-60 per cent of the hepatic parenchyma. Microscopically, the cysts were uni- or multilocular and lined by low cuboidal or flattened epithelial cells. The hepatic parenchyma around cysts showed pressure atrophy, necrosis, engorged sinusoids or other blood vessels or even haemorrhages, mild to extensive fatty or vacuolar degenerative changes and occasionally proliferation of biliary ducts.

Follicle characteristics of non-woolly Indian goats.

Skin follicular studies of four non-woolly Indian goat breeds are reported. The number of primary follicles ranged from 2 to 14 mm-2 with an overall mean of 6.40 +/- 0.22. Secondary follicles per mm2 ranged from 1 to 23 with an overall mean of 9.48 +/- 0.55. The secondary/primary follicle ratios (S/P) for Black Bengal, Jamnapari, Barbari and Sirohi goats were 1.57 +/- 0.21, 1.15 +/- 0.16, 1.61 +/- 0.21 and 2.04 +/- 0.21, respectively, with an overall mean of 1.59 +/- 0.99. The corresponding values for the total follicles per mm2 for the four breeds were 16.83 +/- 1.39, 15.86 +/- 1.08, 17.66 +/- 1.41 and 13.19 +/- 1.41 with an overall mean of 15.88 +/- 0.66. Per cent primaries were lowest in Sirohi and highest in Jamnapari goats. Analysis of variance revealed significant differences between breeds for the number of primary follicles and S/P ratio. Sex differences and the interaction between breed x sex were not significant for any of the follicle traits studied. On the basis of follicle characteristics the non-woolly short-haired goats offer a reasonable scope for crossing with fibre goats, and Sirohi goats possibly have better skin quality for leather conversion than other goat breeds studied.

Follicle and fibre characteristics of Indian pashmina goats.

The hair follicle and fibre characteristics of Cheghu and Changthangi pashmina goats are reported. The average numbers of primary follicles mm-2 were 6.80, 5.20, 5.67 and 4.00; secondary follicles 42.40, 27.10, 38.56 and 29.67; secondary to primary follicle ratio 6.46, 5.78, 6.79 and 7.60; total follicle population 49.20, 32.30, 44.22 and 33.67 in Cheghu females, Cheghu males, Changthangi females and Changthangi males, respectively. The breed difference was not significant for any of these traits, but the sex difference was significant for a number of secondary follicles and total density. Study of fleece characteristics of Cheghu goats in a 1 cm2 area of five body regions, namely, neck, shoulder, mid-side, flank and abdomen, showed that females had finer fibres (12.06 to 12.12 micron) than males (13.04 to 13.64 micron) in all regions. The pashmina length, pashmina diameter and fibre thickness ratio were significantly different between sexes and between body regions but hair/pashmina ratio and pashmina yield were significant for sex and body regions, respectively.

Haematological studies on Indian pashmina goats.

Haematological studies were conducted on 61 clinically normal pashmina producing goats of the Cheghu breed, acclimatised to the temperate, humid climatic condition of Mukteswar, about 2400 m above sea level. The experimental goats comprised four age groups (birth to one month, six to nine months, three to five years and six to 10 years) of both sexes. The overall values, irrespective of age and sex, for the parameters examined were: red blood cells, 14.17 +/- 1.96 X 10(12) litre-1; haemoglobin, 7.46 +/- 0.79 g dl-1; packed cell volume, 0.31 +/- 0.04 litres litre-1; mean corpuscular volume, 21.62 +/- 2.46 fl; mean corpuscular haemoglobin concentration, 23.72 +/- 1.80 g dl-1; mean corpuscular haemoglobin, 5.11 +/- 0.67 pg; erythrocyte sedimentation rate, 0.00 mm at one hour; plasma protein 6.58 +/- 0.78 g dl-1; icterus index, 9.15 +/- 2.92 units; white blood cells 12.26 +/- 2.66 X 10(9) litre-1; absolute count of lymphocytes, 4.62 +/- 1.40; neutrophils, 5.91 +/- 2.84; monocytes, 0.38 +/- 0.15; eosinophils, 0.32 +/- 0.17 and basophils, 0.05 +/- 0.05 (X 10(9) litre-1). The sex of the animal did not affect the haematological parameters but the effect of age was evident. In newborn kids the haemoglobin, mean corpuscular haemoglobin and packed cell volume values were higher and the total leucocyte count was lower than in other age groups. As the kids grew older lymphocyte numbers decreased while neutrophils increased.

Paragonimus westermanni in tigers (Panthera tigris) in India.

Paragonimus westermanni was found in the lungs of two young tigers (Panthera tigris). Cysts were focally distributed in the lungs, initiating an emphysematous response together with extensive fibrosis in the vicinity of the cysts.

Pathological evaluation of Polystichum squarrosum (D. Don) fern in laboratory rats.

Polystichum squarrosum fern fed (30% w/w) rats showed moderate mortality, decrease in body weight, less body fat and splenomegaly. On post-mortem examination, significant gross lesions were not seen in sacrificed animals. Histopathologically, Polystichum fed rats showed dilated Virchow Robin's space in brain, mild to moderate vascular changes likeoedema, engorgement of blood vessels and haemorrhages in most of the visceral organs, interstitial pneumonia in lungs, focal necrosis and generalised vacuolative degenerative changes in liver, more haemosiderin deposition and presence of higher number of megakaryocytes in spleen, shrunken glomeruli, more peri-glomerular space and more number of glomeruli per microscopic field in kidneys, focal hyperplasia of urinary bladder and moderate to marked depletion of germinal epithelium and spermatids in seminiferous tubules of testes. Pathologically, progressive changes were observed only in liver, urinary bladder and testes on 180 days post feeding (DPF). One fern fed rat sacrificed on 135 DPF showed hepatic tumour which was diagnosed as hepatocellular carcinoma. The results showed that P. squarrosum produced almost comparable pathological changes/preneoplastic lesions as reported in bracken fern fed animals. Long term exposure studies (i.e. 2 yrs) are desired.

Biochemical and histological changes in blood, erythrocytes and tissue of rats on feeding Dryopteris juxtaposita fern.

Biochemical and histological alterations in erythrocytes, liver, kidney and brain of rats fed 30% level of D. juxtaposita fern powder were studied. Significant increase in GSH and decrease in lipid peroxidation, acetyl choline estrase and catalase of RBCs was observed. Significant increase in lipid peroxidation, ATPases and decrease in GST on 80 days post feeding in liver, kidneys and brain and acetyl choline estrase in brain was observed as compared to control. Histopathological studies indicated mild vascular changes in lungs, degenerative changes in testes, focal necrosis in liver and villous atrophy or hyperplasia of lining epithelial cells and hypersecretory activity in intestinal glands. Toxic effect of Dryopteris on rats was due to membrane alterations and oxidative stress and degenerative and vascular microscopic pathological changes.

Experimental infection of mice with Taenia taeniaformis eggs from cats--course of infection and pathological studies.

Cysticercus fasciolaris, the larval form of Taenia taeniaformis is commonly encountered in rodents most often in mice and rats through contaminated feed and bedding materials. The infection is asymptomatic and is considered harmless, but its presence in the laboratory mice/rats could lead to misinterpretation of results for biological experiments. The course of infection and pathogenesis of induced C. fasciolaris was studied in Swiss albino mice. The number of established cysts were not significantly different during the course of infection. The mean diameter of the cysts and the metacestode were significantly different during the course of infection reaching a maximum size of 8.1 +/- 2.2 mm and 80.4 +/- 20.2 mm, respectively on 45 DPI. Histopathologically, on 15 DPI, the duodenum of the affected mice revealed cross sections of early larval stage of C. fasciolaris. On 30 and 45 DPI, the liver showed tract of migration of C. fasciolaris larvae with a thick zone of inflammatory reaction and encapsulation against mature larvae in liver. The routine spontaneous Cystucercus infection is clinically asymptomatic in these animals and is considered harmless. The present experimental infection also followed the same course resulting only in asymptomatic colonisation of the parasites.

Ultrastructural studies on interaction of infectious bursal disease virus (IBDV) and aflatoxin B1 on chick embryo fibroblast cell culture.

Light and electron microscopic evaluation of chick embryo fibroblast (CEF) cell culture inoculated with graded doses (0.25, 2.5 and 25 micrograms/ml medium) of aflatoxin B1 with and without infectious bursal disease virus (IBDV) was undertaken. The light microscopy revealed degeneration, detachment and necrosis of fibroblasts and multiple plaques formation in IBDV infected group without and with (0.25, 2.5 micrograms) aflatoxin B1. The cultures infected with virus, with or without 25 micrograms aflatoxin B1 showed complete detachment from glass surface. Electron microscopy of these cultures showed marked pyknotic or bizarre shaped nuclei, pronounced degenerative changes in the rough endoplasmic reticulum (RER), mitochondria and the presence of multiple vacuoles in the cytoplasm. The viruses were spherical, arrayed, complete, generally closer to nuclei and RER and indistinctly membrane bound. The viruses were either localised or scattered in the cytoplasm. Cultures containing 25 micrograms aflatoxin B1 without or infected with virus showed marked necrosis of cells. In latter group only a few viruses were seen either in infected cells or free in culture. Control cultures failed to show cytopathic changes as observed in the other three groups.

Characterization of toxin from cheilanthes fern and its effect on lymphocyte proliferation and DNA fragmentation.

Thin layer chromatography of aqueous extract of whole Cheilanthesfarinosa fern indicated the presence of ptaquiloside or ptaquiloside like compound, coinciding Rf values with that of Pterosin B standard. HPLC analysis revealed the presence of 26.3 mg/kg ptaquiloside. In vitro studies of the aqueous extract on lymphocyte culture revealed a correlation between stimulative indices and concentration of aqueous extract. Stimulation in lymphocyte proliferation was in order of bracken > cheilanthes > ConA> ptaquiloside standard. On incubation of lymphocyte with aqueous extract of ferns, no DNA damage was observed in isolated DNA.

Effect of bracken (Pteridium a quilinum) and dryopteris (Dryopteris juxtaposita) fern toxicity in laboratory rabbits.

Experimental studies with Bracken and Dryopteris ferns @ 25% concentrate ration mixture were conducted in rabbits. Fern fed rabbits showed progressive anaemia, leukopaenia, lymphopaenia and relative heterophilia. Significant elevations in serum enzymes like serum glutamate oxaloactate transminase (SGOT), serum glutamate pyruvate transminase (SGPT), alkaline phosphatase (ALP), urea and creatinine levels were seen. Histopathologically, rabbits showed mild to moderate vascular changes in most of visceral organs, vacuolar degenerative changes in hepatocytes, hypersecretory activity in intestine, presence of casts in renal tubules and degenerative changes in renal tubular lining epithelial cells. Dryopteris fed rabbits showed somewhat more severe degenerative and vascular changes in different intervals. A low level of toxic principle ptaquiloside was detected in Bracken and Dryopteris ferns by thin layer chromatography (TLC) and high-pressure liquid chromatography (HPLC) methods.

Characterisation of fowl adenoviruses from chickens affected with infectious hydropericardium during 1994-1998 in India.

In the present study characterisation has been done for six group I fowl adenoviruses (FAV) isolated from outbreaks of infectious hydropericardium (IHP) of chickens that occurred in different states/regions of India during the years 1994-98. These six viruses were identified as FAV serotype 4 by virus neutralisation and restriction endonuclease analyses. Antigenic analyses of the viruses revealed close relationship (R-values 0.93-0.96). Under the experimental conditions, we have been able to induce IHP using FAV serotype 4 isolate AD: 411 and were also able detect FAV antigens in myocardial tissues by immunofluorescence assay (a new observation), an indication that IHP causing FAV serotype 4 strain replicate in myocardial tissue. Restriction endonuclease analysis of the viral genomes (approximately 46 Kb), using Hind III, Sma I, Xba I, Bam HI, Pst I and Dra I produced identical genetic profiles. Pst I and Bam HI profiles for these six vitus isolates were identical to those published earlier for an IHP causing Pakistani FAV serotype 4 isolate KR31. The identical genetic profiles of viruses, chronology of the outbreaks of IHP in Pakistan during 1989 onward and later in Jammu and Kashmir, India (1994), suggest that FAV serotype 4 isolates involved in outbreaks of IHP in India had probably spread from Pakistan. In order to prevent further spread and economic losses due to IHP in India, based on the antigenic relatedness data in this paper, any one of the six studied FAV serotype 4 isolates can be used as a candidate for mass production of CEH culture based killed vaccine.