Unravelling the genomic origins of lumpy skin disease virus in recent outbreaks.

Lumpy skin disease virus (LSDV) belongs to the genus Capripoxvirus and family Poxviridae. LSDV was endemic in most of Africa, the Middle East and Turkey, but since 2015, several outbreaks have been reported in other countries. In this study, we used whole genome sequencing approach to investigate the origin of the outbreak and understand the genomic landscape of the virus. Our study showed that the LSDV strain of 2022 outbreak exhibited many genetic variations compared to the Reference Neethling strain sequence and the previous field strains. A total of 1819 variations were found in 22 genome sequences, which includes 399 extragenic mutations, 153 insertion frameshift mutations, 234 deletion frameshift mutations, 271 Single nucleotide polymorphisms (SNPs) and 762 silent SNPs. Thirty-eight genes have more than 2 variations per gene, and these genes belong to viral-core proteins, viral binding proteins, replication, and RNA polymerase proteins. We highlight the importance of several SNPs in various genes, which may play an essential role in the pathogenesis of LSDV. Phylogenetic analysis performed on all whole genome sequences of LSDV showed two types of variants in India. One group of the variant with fewer mutations was found to lie closer to the LSDV 2019 strain from Ranchi while the other group clustered with previous Russian outbreaks from 2015. Our study highlights the importance of genomic characterization of viral outbreaks to not only monitor the frequency of mutations but also address its role in pathogenesis of LSDV as the outbreak continues.

Hemato-biochemical and ultrasonographic evaluation of hepatic lipidosis in dairy buffaloes.

Around 60% dairy animals developed moderate to severe hepatic lipidosis at the time of parturition or during early lactation stage. Most of clinician suspect the hepatic lipidosis during above time window only. However, negative energy balance or feeding of high concentrate diet can lead to hepatic lipidosis at any phase of life. The aim of the present study was to evaluate the potential for diagnosis of hepatic lipidosis by means of hemato-biochemical parameters and ultrasonography of the liver at any stage of life. Here, ultrasonographic back fat thickness measurement was correlated with ultrasonographic features of hepatic lipidosis. A total 60 buffaloes were included under the study and sampled for hematological and biochemical parameters. Hematological parameters did not exhibit any significant difference between healthy and hepatic lipidosis-affected buffaloes. Biochemical parameters like beta hydroxy butyric acid, non esterified fatty acid, aspartate amino transferase, gamma glutamyl transferase and alkaline phosphatase revealed a significant increase, while triglyceride, cholesterol, and glucose declined significantly in hepatic lipidosis-affected buffaloes. Total protein, albumin, and total bilirubin levels did not exhibit any significant difference. Based on ultrasonographic findings, the hepatic lipidosis-affected buffaloes were further sub divided into mild, moderate, and severe groups. Portal vein diameter and depth of portal vein were also estimated in current study. Ultrasonographic examination could diagnose 53.33% hepatic lipidosis cases in buffaloes. Among it, 37.50% buffalo had mild hepatic lipidosis, 33.33% had moderate hepatic lipidosis, and 29.16% had severe hepatic lipidosis. Depth of portal vein significantly increased in hepatic lipidosis cases. However, portal vein diameter exhibited a non-significant difference in mild, moderate, and severe groups of hepatic lipidosis. Back fat thickness also revealed a non-significant difference in mild, moderate, and severe hepatic lipidosis. Above study indicate that B mode ultrasonography of the liver can be employed to differentiate various grades of hepatic lipidosis in buffaloes. Biochemical parameters like NEFA, BHBA, AST, GGT, ALP, TG, cholesterol, and glucose can be helpful to screen the hepatic lipidosis at farm level.

Anti-piroplasmic activity of novobiocin as heat shock protein 90 inhibitor against in vitro cultured Theileria equi and Babesia caballi parasites.

Theileria equi and Babesia caballi are the causative agents of equine piroplasmosis (EP). Currently, imidocarb dipropionate (ID) is the only available drug for treating the clinical form of EP. Serious side effects and incomplete clearance of infection is a major drawback of ID. Heat-shock proteins (Hsp) play a vital role in the life cycle of these haemoprotozoans by preventing alteration in protein conformation. These Hsp are activated during transmission of EP sporozoites from the tick vector (poikilotherm) to the natural host (homeotherm) and facilitate parasite survival. In the present study, we targeted the heat shock protein 90 (Hsp-90) pathway of T. equi and B. caballi by using its inhibitor drug - novobiocin. Dose-dependent efficacy of novobiocin on the growth of T. equi and B. caballi was observed in in vitro culture. Additionally, we examined dose-dependent cell cytotoxicity on host peripheral mononuclear cells (PBMCs) and haemolytic activity on equine red blood cells (RBC). In vivo organ toxicity of novobiocin was also assessed in a mouse model. The IC50 (50 % inhibitory concentration) value of novobiocin against T. equi and B. caballi was 165 µM and 84.85 µM, respectively. Novobiocin significantly arrested the in vitro growth of T. equi and B. caballi parasites at 100 µM and 200 µM drug concentration, respectively. In vitro treated parasites had distorted nuclear material and showed no further viability. Based on the equine PBMCs and RBC, the drug was found to be safe even at 1000 µM concentration and the CC50 (50 % cytotoxicity concentration) values were 11.63 mM and 261.97 mM. Very high specific selective index (SSI) values (70.47 and 1587) were observed for equine PBMCs and RBC, respectively. Organ-specific biochemical markers and histopathological examination indicated no adverse effect of the drug at a dose rate of 50 mg kg body weight in the mouse model. The results demonstrate the growth inhibitory effect of novobiocin against T. equi and B. caballi parasites and its safety for host cell lines with very high SSI. Hence, it can be inferred that the Theileria/Babesia Hsp-90 family are potential drug targets worthy of further investigation.

Metagenomic of clinically diseased and healthy broiler affected with respiratory disease complex.

In recent past, the respiratory infection has emerged as a great challenge to the poultry farmers. Various pathogens including Avian pneumovirus (APV), Avian influenza virus (AIV), Infectious bronchitis virus (IBV) and Newcastle disease virus (NDV), Avibacterium paragallinarum, Ornithobacterium rhinotracheale (ORT), Mycoplasma synoviae (MS), Mycoplasma gallisepticum (MG) and Avian pathogenic Escherichia coli (APEC) are involved in the respiratory disease complex in birds [1], [2] (Bradbury, 1984; Roussan et al., 2008). Hence, respiratory disease complex is the most serious disease affecting to poultry and causes heavy economic losses in the poultry industry worldwide [3] (Murthy et al., 2008). In recent years, metagenomics is powerful analyzing tool for detection of pathogens directly from clinical samples without any prior knowledge of the organism in a given sample [4], [5] (Schuster, 2008; Pereira et al., 2010). High throughput Next-Generation-Sequencing technology was used for sequencing the isolated genomic DNA. These data provides an insight about taxonomic and functional status of microorganisms responsible for causing respiratory infection in broiler. The data of these metagenome are available in the BioSample Submission Portal as Bioproject PRJNA339659 and SRA accession number SRR5997823, SRR5992854, SRR6037376, SRR6024702, SRR6012248 and SRR6008913.

Immunohistochemical, histopathological study and chemoprotective effect of Solanum nigrum in N-nitrosodiethylamine-induced hepatocellular carcinoma in Wistar rats.

BACKGROUND AND AIM: Cancer is a devastating disease with a severe impact on the physical and psychological well-being of patients. Hepatocellular carcinoma (HCC) has been reported in various species of animals including dogs, cats, sheep, and pigs. The present study aimed to study the immunohistochemical and histopathological changes andchemoprotective effect of aqueous and alcoholic extracts of Solanum nigrum on N-nitrosodiethylamine (NDEA)-induced HCC rat model. MATERIALS AND METHODS: Eighty-two male Wistar rats of 15 weeks of age weighing 200-250 g were selected for the experiment. They were randomly divided into ten groups. Group I served as normal control consisted of healthy rats. HCC was induced in Group II, IV, V, VI, VII, and X rats using NDEA as inducing agent followed by phenobarbitone as a promoter for 16 weeks. Group II rats were kept untreated as HCC control. Group III rats were kept as vehicle control (0.05% Sodium bicarbonate). Group IV and V rats were treated with aqueous extract of S. nigrum at 200 mg/kg and 400 mg/kg, respectively, and Group VI and VII rats were treated with an alcoholic extract of S. nigrum at 200 mg/kg and 400 mg/kg, respectively, daily orally for 28 days. Group X rats were treated withsorafenib as reference drug at a dose of 11.4 mg/kg daily orally for 28 days. Group VIII and IX rats were kept as aqueous and alcoholic extract control for studying the effect of the same on normal rats. Liver samples were collected to study the gross and histopathological lesions and the activity of cleaved caspase-3 and chemopreventive effect of aqueous and alcoholic extracts of S. nigrum on HCC. RESULTS: The liver sections of rats from HCC control (Group II) showed loss of lobular architecture, necrosis, fatty change, enlarged and darkened nuclei with variable size, dilatation of hepatic sinusoids with Kupffer cell hyperplasia, dilatation and proliferation of bile duct, and intranuclear vacuoles and also showed the presence of more than one nucleolus. Administration of alcoholic extract of S. nigrum and sorafenib to NDEA/phenobarbital-treated rats reduced the severity of lesions in the liver. Immunohistochemical analysis of liver sections for caspase-3-positive cells of hepatic cancer-induced group showed immunoreactivity to rarely few. The immunoreactivity of the hepatocytes treated with a higher dose of alcoholic extract of S. nigrum was limited and was comparable to a standard drug, sorafenib. CONCLUSION: Oral administration of aqueous and alcoholic extracts of S. nigrum for 28 days showed significant rejuvenation in the structure of the liver in the histopathological section in a dose-dependent manner in rats.

Hematobiochemical changes in ehrlichiosis in dogs of Anand region, Gujarat.

AIM: The present research work was undertaken to study the diagnostic importance of hematobiochemical changes in naturally occurring ehrlichiosis in dogs of Anand region, Gujarat irrespective of their age, breed, and sex. MATERIALS AND METHODS: Blood samples from a total of 29 dogs of Anand region of Gujarat state were screened for detection of anti-Ehrlichia canis antibodies using Immunocomb(®) rapid diagnostic kit (Biogal Galed Laboratories, Israel) and subjected to estimation of hematobiochemical parameters by auto hematology analyzers at College of Veterinary Science and Animal Husbandry, Anand. Statistical analysis, interpretation and comparison of hematobiochemical changes with scientific literature was carried out in order to understand the pathophysiology of the disease. RESULTS: Of 29 dogs, 18 were positive for naturally occurring ehrlichiosis based on the presence of anti-E. canis antibodies while 11 were negative. Haematology evinced that the mean values of hemoglobin, total erythrocyte counts, platelet count and packed cell volume in dogs with ehrlichiosis decreased significantly (p<0.01) in comparison to healthy dogs. Among differential leucocyte count, mean values of lymphocytes decreased, neutrophils increased, eosinophils decreased and basophils decreased significantly (p<0.05) in dogs with ehrlichiosis in comparison to healthy dogs while statistically non-significant (p>0.05) difference was observed in values of monocytes in dogs with ehrlichiosis and healthy dogs. Among various red blood cells indices, the mean values of mean corpuscular hemoglobin concentration increased significantly (p<0.01) in dogs with ehrlichiosis in comparison to healthy dogs. Serum biochemistry revealed significant (p<0.01) increase in serum glutamic pyruvic transaminase, serum glutamic oxaloacetic transaminase and creatinine levels as well as decrease in total protein levels in dogs with ehrlichiosis as compared to healthy dogs. CONCLUSION: Clinical importance of hematobiochemical changes in 18 natural cases of ehrlichiosis in dogs of Anand region, Gujarat irrespective of their age, breed and sex is discussed, which would aid new insights in diagnosis and therapeutic management.

Comparison of molecular and microscopic technique for detection of Theileria annulata from the field cases of cattle.

AIM: Tropical theileriosis is fatal hemoprotozoal disease of dairy animals caused by Theileria annulata. The aim of the present study was to detect the T. annulata and comparison of results of molecular and microscopic techniques. MATERIALS AND METHODS: A total of 52 blood samples were collected from the cattle suspected for theileriosis across the Banaskantha district. All the samples were screened for theileriosis using Giemsa's staining technique and polymerase chain reaction (PCR). RESULTS: Total of 17 (32.69%) and 24 (46.15%) samples were found positive for theileriosis by microscopic examination and PCR test, respectively. It revealed that the study area is endemic for theileriosis, and the microscopic technique has 70.83% sensitivity and 100% specificity with respect to PCR technique. CONCLUSION: It may be concluded from the present study that the PCR is comparatively sensitive technique than microscopic examination and may be recommended to use in the field for screening of theileriosis in the study area, where a high prevalence of diseases have been reported due to intensive dairy farming.

Thyroid hormone modulates food intake and glycemia via ghrelin secretion in Zucker fatty rats.

Hyperthyroidism is known to increase food intake and central administration of thyroid hormone shows acute orexigenic effects in rodents. We investigated whether T3 influences appetite and glucose homeostasis by modulating circulating ghrelin, an important orexigenic hormone, in Zucker fatty rats. The acute anorectic effects of T3 and ghrelin mimetic MK-0677 were studied in rats trained for fasting induced food intake. The serum concentration of T3, ghrelin, glucose, triglycerides, and liver glycogen were estimated. The involvement of sympathetic nervous system was evaluated by conducting similar experiments in vagotomized rats. T3 increased food intake and glucose in rats over 4 h, with increase in serum T3 and decrease in liver glycogen. T3 treatment was associated with increase in serum ghrelin. An additive effect on appetite and glucose was observed when T3 (oral) was administered with central (intracerebroventricular) administration of a ghrelin mimetic, MK-0677. Ghrelin antagonist, compound 8a, antagonized the hyperglycemic and hyperphagic effects of T3. In vagotomized rats, T3 did not show increase in appetite as well as glucose. Serum ghrelin levels were unchanged in these animals after T3 treatment. However, T3 showed increase in serum triglyceride levels indicating its peripheral lipolytic effect, in vagotomized as well as sham treated animals. To conclude, acute orexigenic and hyperglycemic effects of T3 are associated with ghrelin secretion and activity. This effect seems to be mediated via vagus nerves, and is independent of glucoregulatory hormones.

Epidemiological survey of equine influenza in horses in India.

A highly contagious virus infection in horses, influenza is the single most important equine respiratory disease in the world. This paper presents details of a one-year study (1 June 2008 to 31 May 2009) to determine the prevalence of equine influenza in the horses of Gujarat State in India. The prevalence of equine influenza A/equi-2 was 12.02%, but none of the samples were positive for equine influenza A/equi-1. The prevalence of equine influenza (A/equi-2) was 15.38%, 11.94%, 10.18%, and 9.09% in horses of the Kathiyawari breed, a non-descript breed, the Marwari breed and the Indian Thoroughbred breed, respectively. The highest prevalence of influenza was observed in yearlings (17.48%) and prevalence was at its highest in the month of April (28.89%). The prevalence rate in males, females and geldings was 11.95%, 10.38% and 8.47%, respectively. The mortality rate and case fatality rate were 1.28% and 10.64%, respectively.

Study on tobacco use and awareness among marginalized children.

BACKGROUND: The current study entailed a survey of children from the lower socioeconomic strata of rural and urban regions of the states of Maharashtra and Assam who are vulnerable to tobacco usage. More than 1700 children were checked for precancerous lesions and 1004 were surveyed for tobacco habits and awareness. AIMS: The objective of the survey was to determine and report on all the variant factors affecting the use of tobacco among the underprivileged children population. The aim of the clinical check-up was to detect precancerous lesions in the tobacco-using children at an early treatable stage. MATERIALS AND METHODS: Awareness lectures and ENT camps were conducted at 12 organizations/community centers. A cross-section of children were interviewed to understand tobacco use among them. All the children were screened for precancerous lesions. Children with suspicious oral lesions were sent for further evaluation at a nearby diagnostic cancer facility. The survey was conducted by trained social workers. RESULTS: The percentage of tobacco users in urban Mumbai was quite low at 4.8% compared with rural Kasara (36%) and Assam (76%); and 74.6% of the children were aware that tobacco use was dangerous and harmful to health. The average age of initiation was 9 years. Out of the 1004 children surveyed, 253 were tobacco users and 79% were males. Of the 1700 children screened, 23.5% presented with precancerous oral lesions. CONCLUSION: This study addresses the tobacco habits of a typical sample of marginalized children in India and the need for effective interventions aiming at reducing the burden of tobacco-related cancers by controlling at the point of initiation.

Novel oxazole containing phenylpropane derivatives as peroxisome proliferator activated receptor agonists with hypolipidemic activity.

alpha-Alkoxy arylpropanoic acids containing 2-phenyloxazole-4yl-alkyl moiety are found to be potent hypolipidemic agents. These compounds were potent activators of the peroxisome proliferator activated receptor gamma (PPARgamma), with moderate PPARalpha activity and known to cause adverse effects such as weight gain and edema, which are essentially attributed to PPARgamma activation. Although extensive work has been done on the phenylpropanoic acid class of compounds, other phenyl propane derivatives such as alcohols, amines, ethers etc. have not received much attention. In order to develop predominant PPARalpha agonists as hypolipidemic agents with minor chemical modifications on compound III, we have synthesised few (2S)-ethoxyphenylpropane derivatives containing a 2-phenyl-5-methyloxazole-4ylalkoxy moiety of the general formula IV and evaluated by PPARalpha and gamma transactivation assay in conjugation with in vivo studies in male Swiss albino mice model. Compounds 3c and 3d showed the desired predominant PPARalpha activity and excellent tryiglyceride reduction in vivo and were selected as lead compounds for further development as hypolipidemic agents.

Peritumoral apparent diffusion coefficient as a metric of response in patients with recurrent glioblastoma multiforme treated with bevacizumab and irinotecan.

Bevacizumab and irinotecan have shown promising results in patients with recurrent glioblastoma multiforme (GBM), which traditionally carries a poor prognosis after first-line therapies have been exhausted. Retrospectively documenting the short-term effects of this chemotherapeutic regimen on recurrent GBM, as evidenced by comparative magnetic resonance images obtained two weeks prior to, and one-month following initiation of treatment, we hypothesize that peritumoral apparent diffusion coefficient (ADC) values will decrease on post-treatment scans. Brain MR data were collected from August 2005 to December 2006, in which post-contrast T1-weighted images demonstrated measurable enhancement or GBM tumor mass. Pre- and post-treatment MR images for ten consecutive patients were collected, each having failed temozolomide and radiation therapy. Pre- and post-treatment recurrent GBM bulk tumor and peritumoral T2 signal abnormality were measured in three dimensions. Diffusion of peritumoral T2 signal abnormality was evaluated on pre- and post-treatment ADC. All patients witnessed a significant decrease in tumor bulk ranging from 15.3% to 96.7% with a mean reduction of 48.2%, having received an average of two cycles of chemotherapy. FLAIR images demonstrated a mean volumetric reduction in peritumoral T2 signal abnormality of 44.3%. ADC measurements demonstrated an average reduction in peritumoral ADC of 20.6%, which was statistically significant (p-value < .005). Recurrent GBM tumor bulk demonstrated a 48.2% mean reduction, with corresponding decrease in peritumoral ADC values of 20.6%, suggesting that ADC may represent a valuable metric in the evaluation of the chemotherapeutic response of recurrent GBM, when treated with bevacizumab and irinotecan.

Up-regulated p75NTR neurotrophin receptor on glial cells in MS plaques.

OBJECTIVE: To investigate the expression of the neurotrophin receptor p75NTR on glial cells within MS plaques. BACKGROUND: In recent studies on the pathogenesis of MS white matter plaques, we found large populations of inflammatory and resident glial cells, including oligodendrocytes undergoing cell death, and identified increased expression of Fas receptor and ligand death pathway signaling molecules on the same glial cell types. In another study, the p75NTR was shown to induce apoptotic death of maturing oligodendrocytes when exposed to NGF in vitro. METHODS: We used immunohistochemistry and in situ reverse-transcription PCR to detect p75NTR expression on inflammatory and resident glial cells in MS plaques and used TUNEL staining for fragmented DNA to detect cell death. RESULTS: Up-regulated p75NTR messenger RNA and protein were demonstrated in both oligodendrocytes and microglia/macrophages in MS plaques but not in control white matter. However, only a fraction of p75NTR expressing oligodendrocytes was also stained by TUNEL. CONCLUSIONS: Glial cell expression of p75NTR receptor is up-regulated during MS plaque formation. The exact role of this receptor in glial cell death and/or survival in MS remains to be elucidated.

Involvement of the CD95 (APO-1/Fas) receptor/ligand system in multiple sclerosis brain.

Immunohistochemical methods were used to search for Fas receptor/Fas ligand system involvement in multiple sclerosis (MS) white matter brain lesions. We found large numbers of Fas ligand (Fas-L)-bearing cells present in two acute lesions and 12 of 16 chronic MS lesions, and very few positive cells in non-inflammatory controls. Four of six brains from non-MS neuropathologic conditions associated with inflammation and white matter disease were, however, also positive for Fas-L. Double staining with cell-specific markers revealed that the pattern of ligand-positive cells in chronic MS lesions was complex and composed of several different cell types which were primarily resident glial cells with a small overlay of macrophages. Fas/APO 1 (CD95) receptor expression in MS tissue was also evaluated and marked upregulation of the receptor was found. In addition, Fas receptor was induced, but to a lesser extent, in numerous control brains. The observations that TUNEL-positive dying cells were present in MS lesions and showed excellent co-localization with Fas-L, indicate that the Fas death system may contribute to plaque pathogenesis and could lead to the development of a new category of therapeutic agents for MS.

A teratocarcinoma-derived neurotrophic activity.

In a search for nerve growth factor (NGF) in tissue extracts of a murine transplantable teratocarcinoma that harbours immature neural tissue in abundance, a trypsin-sensitive and heat labile neurotrophic activity was identified. The final protein fraction obtained by cation exchange chromatography contained five proteins with mol. wts ranging from 52 to 72 kD. It supported the growth and differentiation of immature neurones in neonatal rat cerebellar cultures but had no effect on embryonic chick dorsal root ganglia which are the classical targets for NGF.

A study of continuous bacillaemia in borderline and lepromatous type of leprosy.

The present work has established beyond doubt that bacillaemia occurs practically in all cases of borderline and lepromatous leprosy. The number of bacilli in the blood is significantly high, to the tune of 4,000 bacilli per ml. Even so, it does not produce any adverse symptoms of septicaemia. Special emphasis was laid in the present study for determining the continuity or otherwise of bacillaemia. Some of the patients had continuous bacillaemia according to the criteria fixed in the present study. It was also found that the number of bacilli discharged into the blood bears no relationship with the bacillary load in the body as assessed by skin and nasal smears.