Isolation and molecular characterization of fowl adenovirus from inclusion body hepatitis cases in Western India during 2019-2021.

Background: Inclusion body hepatitis (IBH) resulted in a substantial economic loss in Western India during 2019 to 2021. Aims: The study aimed to characterize fowl adenovirus (FAdV) from field outbreaks. Methods: The study was conducted on 290 liver samples from 66 poultry flocks. The samples were subjected to histopathology and molecular detection, followed by phylogenetic typing of the partial hexon gene of the virus. Results: Spiking mortality (14%) was recorded from day 21 to day 35 with peak mortality at the 28th day of age. The necropsy showed a pale and enlarged liver with hemorrhagic and yellowish necrotic foci, accumulation of straw-colored transudate in the pericardial sac which resulted in a flabby appearance of the heart, heart enlargement, and hemorrhages on the spleen, enlarged and congested kidneys. The virus inoculation resulted in stunting and poor feathering with hepatomegaly, hemorrhages and yellowish necrotic foci on the liver as well as greenish discoloration, and kidney swelling in SPF embryonated chicken eggs. Out of 29, 16 liver samples yielded 1219 bp amplicons specific to hexon gene fragments. The sequence and phylogenetic analysis identified 14 isolates as FAdV species E serotype 11 and two as species D serotype 8b. Conclusion: The results indicated that FAdV-8b and FAdV-11 strains are involved in disease outbreaks in western India.

Clinical, hematobiochemical, and pathological findings and therapeutic management of viperine snake envenomation in zebu cattle.

The present investigation was carried out to study the epidemiology, clinical signs, bleeding profile, kidney function, and pathology and to evaluate the therapeutic protocol for viperine snake envenomation in zebu cattle over a period of 14 years. A total of 98 cases of viperine bite were reported with the highest incidence during monsoon months in grazing male zebu cattle. Ascending swelling over the affected limb with lameness or asymmetrical swelling over the face with dyspnea was a consistent clinical finding. Increased bleeding tendency was invariably observed in ailing cattle with major bleeding from the site of the bite. Hematobiochemistry showed neutrophilic leukocytosis, thrombocytopenia, prolonged capillary blood clotting time, and elevation of BUN and creatinine. The characteristic gross pathological lesions observed were widespread petechial to ecchymotic hemorrhages in the lungs, liver, heart, spleen, abomasal mucosa, and intramuscular or subcutaneously at the site of bite. Histopathological examination revealed presence of necrohemorrhagic changes within the liver, heart, kidneys, skeletal muscles, and subcutaneously or intradermally at the site of bite. Diagnosis of viperine snakebite was made based on circumstantial evidence of bite, progressive swelling, and bleeding at the site of bite. The severity was assessed based on prolongation of capillary blood clotting time, thrombocytopenia, and kidney dysfunction. Therapeutic protocol comprising polyvalent anti-snake venom along with supportive therapy for 5-9 days showed encouraging results with 88.30% survival rate. In conclusion, the precise diagnosis of viperine snake envenomation in cattle can be made from history and typical clinical signs while severity and treatment can be monitored based on bleeding profile.

Acaricidal potency of polyherbal spray against Rhipicephalus microplus and Hyalomma anatolicum infestation in cattle.

Acaricidal potential of polyherbal spray (Andropogon citrates, Cymbopogon citratus, Ocimum sanctum, Pinus longifoia, Calotropis procera, Datura stramonium, Aegle marmelos, Ricinus communis, Azadirachta indica, Allium sativum, Carica papaya, Annona squamosa and Pongamia glabra) was assessed against tick infestation in cattle on the basis of measurement of tick count, complete blood count and plasma glucose, total protein, albumin and globulin before treatment and 21 days after treatment. Single application of polyherbal spray over body of 20 randomly selected tick infested cattle revealed significant reduction in mean tick count starting from 3 days post treatment till 21 days post treatment. Highly significant (P < 0.01) increase in total erythrocyte count and packed cell volume was observed in treated cows 21 days after application of spray compared to pre-treatment values indicating the reduction in blood loss due to heavy tick infestation before treatment. Plasma biochemical parameters revealed no significant changes in pre-treatment and post treatment values. The results of present study imply the clinical and haematological improvement in tick infested cattle treated with polyherbal spray and it could be potential product for use in livestock as acaricide.

Identification of Forced Degradation Products of Itopride by LC-PDA and LC-MS.

Degradation products of itopride formed under different forced conditions have been identified using LC-PDA and LC-MS techniques. Itopride was subjected to forced degradation under the conditions of hydrolysis, photolysis, oxidation, dry and wet heat, in accordance with the International Conference on Harmonization. The stress solutions were chromatographed on reversed phase C18 (250×4.6 mm, 5 µm) column with a mobile phase methanol:water (55:45, v/v) at a detection wavelength of 215 nm. Itopride degraded in acid, alkali and oxidative stress conditions. The stability indicating method was developed and validated. The degradation pathway of the drug to products II-VIII is proposed.

Isolation of an unknown compound, from both blood of Bhopal aerosol disaster victims and residue of tank E-610 of Union Carbide India Limited--chemical characterization of the structure.

A total of more than 28 chemical entities/reaction products in the form of gases, vapour and particulate matter were reported from the tank E-610 of methyl isocyanate (MIC) storage tank of Union Carbide India Limited on the night of 2/3 December 1984 in Bhopal. In earlier studies, methyl isocyanate and its trimer, with a few other compounds, were reported in the human victims preserved in deep freeze. Randomly selected samples were analysed by gas chromatograph coupled with mass spectrometer (ITD-800, Finnigan MAT, UK). Four of the cases showed the peaks and fragmentation pattern identified with one of the unidentified compound of molecular weight 269 amu in the Tank Residue, which constituted about 0.2 area per cent on GC-ITD. After isolation by column chromatography and being exposed to characterization, it was identified as a Spiro compound. It was possibly formed by the polymerization of five molecules of methyl isocyanate.

Distribution of malathion in body tissues and fluids.

Six cases of suspected poisoning were studied. The various body tissues and fluids of all the cases were analysed and malathion was found positive. The quantitative analysis was performed using high performance liquid chromatography. The distribution of malathion was studied in lungs, liver, kidneys, spleen, brain, heart, blood, muscles, urine and gastric contents.

GC-NPD and GC-MS analysis of preserved tissue of Bhopal gas disaster: evidence of methyl carbamylation in post-mortem blood.

Twenty-five preserved autopsy blood samples of Bhopal toxic gas exposed victims were analysed by gas chromatography (GC) coupled with either Nitrogen-Phosphorous detector (NPD) or mass spectrometer (MS) for the presence of methyl carbamyl valine in terms of valine methyl hydantoin (VMH). 84% of these samples showed a positive test for VMH on GC-NPD and the identity of the peaks were further confirmed on GC-MS. The concentration of VMH in the gas-affected positive blood samples ranged from 2.56 to 51.28 nanomoles. These results indicate entry of methyl isocyanate (MIC), one of the constituents of the toxic cloud caused by the disaster, into the blood stream of victims who had inhaled gas.

GC-MS identification of MIC trimer: a constituent of tank residue in preserved autopsy blood of Bhopal gas victims.

Based on the external and internal findings of Bhopal gas disaster victims, it was apparent that the gases and particulate matter came out as an aerosol. This was possibly the pyrolysed, reformulated, reconjugated suspension of constituents of the tank E-610 of Union Carbide India Limited, Bhopal, while it was claimed to be methyl isocyanate (MIC) only. It was postulated by the manufacturer of MIC, that the material inhaled by the victims of the Bhopal gas disaster does not cross the lung barrier (UCC press conference on 14th December 1984). It was observed that the more the victims ran, the more aerosol they inhaled and the fatalities were observed in such victims. The tissues, which were preserved in the deep freeze, were randomly selected and analysed by GC coupled with MS (ITD) Finnigan MAT, UK. 14 out of 34 autopsy cases showed MIC trimer peak in extracts of blood. This was one of the constituents of the aerosol and was also located in the tank residue, thereby proving that the trimer had passed the lung barrier.

High performance liquid chromatographic determination of alcohols with reference to body distribution of methanol.

A method has been developed on reverse phase high performance liquid chromatography for simultaneous determination of methyl, ethyl and isopropyl alcohols under refractive index detection using pure water as the mobile phase. A good separation has been achieved between these alcohols. Detector response was linear with a detection limit of 5 mg/100 ml. Recovery studies were performed by adding known amounts of methyl and ethyl alcohols to blood, lung and liver within the range 80-90%. The reproducibility of the results was always greater than 90%. The quantitative distribution of methyl alcohol in postmortem body tissues and fluids has been reported in three cases of poisoning.

High performance liquid chromatographic method for the analysis of organophosphorus and carbamate pesticides.

A method has been developed for the rapid quantitative analysis of organophosphorous and carbamate pesticides using HPLC. Good separation was obtained among the four major groups of organophosphorus pesticides [i.e. aryl phosphorothionate (methyl parathion), alkyl phosphorothionate (malathion), enolphosphate (phosphomidon, monocrotophos, dichlorvos), heterocyclic phosphorothionates (quinalphos)] and carbamates [viz. Carbaryl (Sevin) and Baygon (Dalf)] with a detection limit of 100 ng for all the pesticides. Separation was measured in terms of capacity factor (k') resolution (R) and selectivity factor (alpha ii). The method described can be used for the analysis of biological samples for the presence of organophosphorus and carbamate pesticides in the cases of poisoning. Recovery studies were made in the blood, lung and liver and found to be 85-97% with reproducibility at greater than 95%.