First record of molecular and pathological investigations of physalopterid larvae infection in northern palm squirrel Funambulus pennantii.

Around 100 reported species of Physaloptera commonly infect mammals, reptiles, birds, and amphibians. The identification of Physaloptera species solely on morphological characteristics is difficult, especially in the case of larval and congeneric infections. The present study is an attempt to identify molecularly and to perform phylogeny and pathology of natural Physaloptera larval infection in northern palm squirrels. The molecular confirmation of the recovered parasitic stages was performed by targeting the nuclear 18S rRNA gene sequence. Phylogenetic analysis and evolutionary divergence of the present study isolate with GenBank™ archived Physaloptera sequences were performed. The cysts (containing the larval stages) were subjected to histopathological examination. Morphological identification of the larval stages revealed the presence of pseudolabia, two spines, and a collar-like projection at the anterior end. Histopathology of the cysts revealed transverse sections of parasites in the lumen along with the thickened cystic wall, infiltration of mononuclear cells, fibrous tissue proliferation in the wall, and cellular debris in the cystic lumen. The molecularly confirmed and sequenced present study isolate was submitted to GenBank™ under the accession number LC706442. Blast analysis revealed 96.82-98.64% nucleotide homology of the present study isolate to the GenBank™ archived Physaloptera sequences. The isolate of the present study was monophyletic with Physaloptera sp. and P. praeputialis recovered from the cats of Haryana, India. Also, evolutionary divergence studies revealed no difference among these sequences. The present study evinced the most probable role of the northern palm squirrel Funambulus pennantii as an aberrant or second intermediate host for P. praeputialis.

Phylogenetics and Pathology of Hydatid Disease in Slaughtered Buffaloes of North India.

PURPOSE: Hydatid disease is one of the neglected and challenging (for diagnosis as well as for treatment) parasitic diseases. Along with adverse effect on animal's health leading to production losses, hydatidosis is also associated with huge economic losses. The present study was envisaged with an aim to assess the phylogeny and pathological changes due to natural hydatid cysts in lungs and liver of slaughtered buffaloes in north India. METHODS: A total of 137 slaughtered buffaloes intended for human consumption were screened for the presence of cysts. The retrieved cysts were confirmed molecularly based on the amplification of mitochondrial cytochrome oxidase 1 gene (mtCO1), exhibiting a product size of approximately 446 bp. The samples collected from infected lungs and liver were subjected to histopathological examination. RESULTS: The hydatid cysts were recorded in 25 (18.2%) animals. Phylogenetic analysis revealed the isolated strain to be closely related to Echinococcus granulosus sensu stricto (G1) genotype. The nucleotide diversity (π) obtained was 0.014685, whereas, Tajima's D was negative (- 2.796053), which indicated purifying selection or recent population expansion. Histopathologically, in the infected lungs, fibrosis and inflammatory reaction comprising of mononuclear cells and fibroblasts around the thick coat of granulation tissue were observed. Marked calcified masses and necrosis were also observed in the calcified cysts. However, in case of infected livers, Kupffer cell hyperplasia, degeneration of hepatocytes, fibrosis and inflammatory cells were most commonly observed around the hydatid cysts. CONCLUSION: The findings of the present study are of significant veterinary and medical importance owing to economic and public health impact of G1 genotype of E. granulosus.

Biochemical studies in experimentally Escherichia coli infected broiler chicken supplemented with neem (Azadirachta indica) leaf extract.

AIM: An experimental study was conducted on 192-day-old broiler chicks for evaluating the effect of 10% neem leaf extract (NLE) supplementationon biochemical parameters in chickens experimentally infected with Escherichia coli O78 at 10(7) CFU/0.5 ml at 7 days of age. MATERIALS AND METHODS: The 192-day-old broiler chicks were procured. These chicks were divided into two groups (A and B) containing 96 birds each on the 1(st) day. Diet of all the chicks of Group A was supplemented with 10%NLE in water, whereas chicks of Group B were given feed and water devoid of NLE supplementation throughout the experiment. After rearing for 1 week, chicks of both the groups (A and B) were again divided into two subgroups (Group A into A1 and A2 and Group B into B1 and B2) of 54 and 42 birds, respectively. At the age of 7 days all the chicks of groups A1 and B1 were injected with E. coli O78 at 10(7) CFU/0.5 ml intraperitoneally. Blood samples were collected from six chicks from each group at day 0, 2, 4, 7, 14, 21, 28 days post-infection and serum was separated for biochemical studies. RESULTS: There was a significant increase in serum alanine transaminase (ALT), aspartate transaminase (AST), lactate dehydrogenase (LDH) activities, globulin concentration and a decrease in total protein (TP), albumin concentrations, and alkaline phosphatase (ALP) activity in both the infected groups. However, the changes in biochemical values, i.e., ALT, AST, LDH, ALP, TP, albumin, and globulin wereof lower magnitude in NLE supplemented group suggesting hepatoprotective and cardioprotective effect of NLE. CONCLUSIONS: Fromthe present study, it is reasonable to conclude that significant increase in the value of ALT, AST, LDH, globulin, and significant decrease in the value of ALP, TP, and albumin was of lower magnitude in supplemented infected group (A1) as compared to non-supplemented infected group (B1) suggesting hepatoprotective and cardioprotective effect of NLE.

Pathomorphological and microbiological studies in sheep with special emphasis on gastrointestinal tract disorders.

AIM: The present study was envisaged to elucidate the pathomorphological and microbiological aspects of gastrointestinal tract (GIT) disorders of sheep/lambs. MATERIALS AND METHODS: Samples for research were collected from 12 sheep died with a history of GIT disorders which were brought for post-mortem examination to the Department of Veterinary Pathology, Lala Lajpat Rai University of Veterinary and Animal Sciences, Hisar, for pathomorphological and microbiological examination. RESULTS: Gross pathological changes in various organs noticed were abomasitis, congestion and hemorrhages in intestine; necrotic foci on liver surface; enlarged, hard, and indurated mesenteric lymph nodes, hydropericardium, congestion, hemorrhages and consolidation of lungs and congestion and soft kidneys as the major change. On histopathological examination, there were abomasitis with leukocyte infiltration, enteritis with desquamation of mucosal epithelium and goblet cell hyperplasia, lymphadenitis with depletion of lymphocytes in the germinal center of lymphoid follicle, and splenitis with depletion of lymphocytes in the white pulp. In the liver congestion, degenerative changes in hepatocytes including cloudy swelling, fatty changes, congestion in sinusoids, and dilatation of sinusoids leading to atrophy of hepatocytes. Lungs evidenced edema, congestion, emphysema, serous inflammation, thickening of interlobular septa, fibrinous pleuritis, and peribronchiolar lymphoid follicle formation. Heart revealed sarcocystosis, fibrinous pericarditis, and hyalinization of the myocardium. In kidneys, congestion, focal interstitial nephritis, hyaline degeneration, and coagulative necrosis were seen. For microbiological aspects; cultural isolation was done from samples of liver, abomasum, mesenteric lymph nodes, spleen, heart blood, lungs, and kidneys from the carcasses of sheep/lambs. Escherichia coli was the only bacterium isolated during present studies. E. coli isolates from different tissues of carcasses of sheep/lambs were subjected to in-vitro drug sensitivity testing. Ciprofloxacin, cefixime, polymyxin B, amoxicillin + sulbactam, and amoxicillin + clavulanic acid were the most sensitive drugs followed by amikacin, ofloxacin, ampicillin, co-trimoxazole, and amoxicillin. CONCLUSIONS: From the present study, it is reasonable to conclude that the major etiopathological cause of GIT disorders in sheep was E. coli infection, which causes a pathomorphological effect on various cadaver organs viz. abomasum, intestine, liver, mesenteric lymph nodes, lungs, spleen, kidneys, and heart followed by parasitic infection of Haemonchus contortus.