Ethanolic extract of Azadirachta indica (A. Juss.) causing apoptosis by ROS upregulation in Dirofilaria immitis microfilaria

Dirofilaria immitis is the causative agent of cardiopulmonary dirofilariasis in the Canine family. The aim of the study was to evaluate the efficacy of the ethanolic extract of Azadirachta indica leaves (EEA) against the microfilaria (mf) of D. immitis in vitro. EEA was evaluated for different compound classes through HPTLC. Relative motility, mortality and morphological alterations were observed in the mf after exposure to EEA. The effect of EEA on redox status in the treated mf was evaluated by some key enzymatic and non-enzymatic parameters. An enhanced reactive oxygen species (ROS) level in the treated mf along with altered redox status was evident. DNA fragmentation and terminal-deoxynucleotidyl-transferase dUTP nick end labeling (TUNEL) confirmed apoptosis. In addition, western blotting revealed down-regulation of anti-apoptotic protein and up-regulation of pro-apoptotic proteins. Taken together, the microfilaricidal activity of EEA can be attributed to its capacity to inflict oxidative stress culminating in apoptosis.     

Ultrastructural Alterations in the Gills of Labeo rohita Fingerlings Exposed to Thermal Extremes

This study aimed to determine the cellular alterations in the gill of Labeo rohita exposed to lethal temperature maxima (LT_Max) and lethal temperature minima (LT_Min) by means of transmission electron microscopy (TEM). Acclimation of advanced fingerlings of L. rohita was carried out at 26°C for 30 days. Acclimated fish were subjected to a constant rate of increase or decrease in temperature (0.3°C/min) until the LT_Max and LT_Min values were reached. Dissected gills were processed for TEM, both at the end of acclimation period at ambient temperature (26°C) and at lethal temperatures. Results indicated that at ambient temperature, the gill tissues appeared normal. However, significant changes were observed at lethal temperatures. The gill tissues at lethal temperature maxima showed severely damaged lamellae, with more vacuolated space. At lethal temperature minima, gill tissues showed increased density of mitochondria. Our prima‐facie report indicated that L. rohita exposed to lethal temperatures exhibited marked ultrastructural changes in the gills.     

Surgical treatment of a double aortic arch in a dog

An 8-month-old spayed female Labrador retriever dog was evaluated for regurgitation 6 months after surgery for a suspected vascular ring anomaly. The dog had a history of regurgitation and slow development as a puppy. An initial left-sided exploratory thoracotomy was unsuccessful in identifying and treating a vascular ring anomaly. The dog was subsequently presented to the PennVet Emergency Service for regurgitation. Thoracic radiography showed cranial thoracic esophageal dilation and an esophageal foreign body that was then removed endoscopically. Subsequent computed tomographic (CT) angiography revealed a double aortic arch. A left 4th intercostal space thoracotomy was performed. The smaller left aortic arch and a left ligamentum arteriosum were ligated and transected. The dog recovered uneventfully and was healthy at the 1-month follow-up visit. This is the 5th reported successful surgical correction of a double aortic arch in a dog. Computed tomographic angiography was essential in diagnosis and surgical planning.Key clinical message:Although uncommon, double aortic arches can occur and present a diagnostic and surgical challenge when a persistent right aortic arch is suspected. Computed tomographic angiography provides an accurate preoperative diagnosis and allows for surgical planning.     

Purification of cattle oviduct specific proteins and their effect on in vitro embryo development

The purpose of this study was to determine the effect of oviduct specific proteins as a media supplement for in vitro embryo development in cattle. The proteins were extracted from oviducts of cows and precipitated by ammonium sulfate (30%, 40%, 50% and 60%) followed by dialysis in 50 mM Tris–HCl (pH 7.0) buffer. The dialyzed proteins were fractionated into acidic, basic and neutral fractions using SP sephadex cation exchange and DEAE sephadex anion exchange column chromatography respectively. Cow oviduct specific proteins (cOSPs) constituting all the extracted proteins were used as media supplement in three different concentrations (10, 50 and 100 μg/ml) for in vitro maturation, fertilization and culture (IVMFC) of cow oocytes. Acidic, basic and neutral (unbound) fractions were also used as media supplement in three different concentrations (10, 30 and 50 μg/ml) for IVMFC. Cumulus oocytes complexes were collected from slaughterhouse ovaries, washed thoroughly and cultured in maturation media for 24 h in 5% CO₂ at 38.5 °C with maximum humidity. In vitro matured oocytes were co-incubated with in vitro capacitated sperm in Fert-BO media at 38.5 °C for 18 h in 5% CO₂. The fertilized oocytes were washed and cultured in embryo development media for cleavage. After 40–42 h cleavage was observed and embryos were put in the replacement media for further development. The cleavage rates (%) for cOSPs were observed as 68.24±2.46, 69.28±2.05, 61.77±0.93 and 42.62±1.31 at concentrations of 0, 10, 50 and 100 μg/ml respectively. Rates of blastocyst stage development were 14.49±3.61, 21.17±2.77, 14.66±1.06 and 11.98±1.84. These results indicate that addition of cOSP at10 μg/ml increased blastocyst formation as compared to other concentrations (0, 50 and 100 μg/ml). Although acidic, basic and neutral fractions seemed to have no major effect on cleavage rate, but both acidic and neutral fraction of oviduct specific proteins improved the cleavage rate at 30 μg/ml concentration and basic fraction improved the blastocyst formation at 10 μg/ml concentration.     

Path Analysis of Important Growth Functions of Indian Mustard (Brassica juncea L. Czern and Coss)

Field experiment conducted at the Viswavidyalaya Farm during the winter seasons of 1980—81 to 1983—84 to study the effects of major six productive functions viz. number of inflorescence bearing primary branches, number of inflorescence bearing secondary branches, leaf area indices at flowering, number of seeds/siliqua, number of siliqua/m² and test weight of grains on the grain yields of eight elite varieties of Indian mustard ( Brassica juncea L. Czern and Coss) when sown on three different times revealed that correlations between them were close and positive in all the varieties. Path analysis of the data, further, showed that except the inflorescence bearing primary branches and the seeds/siliqua all other attributes showed high direct influences on grain yield; the maximum direct influence was obtained from number of siliqua/m².     

Polymorphisms in GnRHI and GnRHII genes and their association with egg production and egg quality traits in chicken

1. Two candidate genes, namely, Gonadotropin releasing hormone I (GnRHI) and Gonadotropin releasing hormone II (GnRHII) play pivotal roles in ovulation and egg production in chicken. The objective of this study was to explore polymorphism in these genes and to estimate the effects of polymorphism of these two genes on egg production and egg quality traits in White Leghorn laying hens.

2. Single strand conformation polymorphism followed by sequencing was performed to detect polymorphism in these genes.

3. The coding regions of the GnRHI and GnRHII genes were found to be polymorphic. In the GnRH1 gene, 12 haplotypes were determined, of which the h1 haplotype was predominant and the h5, h9 and h11 haplotypes were the least frequent ones. In the GnRHII gene, eight haplotypes were found, of which the h1 haplotype was the most frequent and the h6 was the least frequent haplotype in the White Leghorn population.

4. The haplogroups of GnRHI had a significant effect on body weight and egg production up to 64 weeks of age, yolk content, Haugh units and egg shell parameters. The h1h2 haplogroup of the GnRHI gene showed the highest egg production, with 211.0 ± 24.3 eggs up to 64 weeks of age, while the highest yolk content and Haugh unit was found in h3h10 haplogrouped birds. The haplogroups of GnRHII had a significant effect on age at sexual maturity (ASM) where the shortest ASM was found in the h1h4 birds (147.3 ± 5.9 d) and the longest ASM was observed in the h1h3 birds (160.6 ± 23.4 d).

5. It was concluded that GnRHI and GnRHII genes are polymorphic and have a significant effect on body weight, egg production and egg quality traits in White Leghorn laying hens.