Evaluation of pheromone-based kit: A noninvasive approach of estrus detection in buffalo.

In view of the silent nature of estrus in buffalo, a noninvasive assay kit has long been felt necessary for easy and effective estrus detection. This study was designed to detect estrus in buffalo using a kit formulated in our laboratory based on pheromone compound. Group I: Urine samples collected at estrus phase and group II: randomly collected urine samples were subjected to the test using the kit. No colour developed (i.e., positive reaction) in estrus urine after adding the kit solution. By contrast, pale and/or dark pink colour developed (i.e., negative reaction) in urine from the proestrus and diestrus buffaloes, respectively. Field evaluation of the kit in groups I and II revealed that 60.87% and 71.43% of urine samples were correctly identified as estrus and nonestrus (i.e., proestrus and diestrus), respectively. Therefore, the first of its kind estrus detection kit formulated based on urinary pheromone can as well be used as a simple device to detect estrus in buffalo.

Gracilaria edulis exhibit antiproliferative activity against human lung adenocarcinoma cell line A549 without causing adverse toxic effect in vitro and in vivo.

In the present study, the antiproliferative potential of various solvent extracts of Gracilaria edulis (GE) was tested against various cancer cell lines. In the A549 lung cancer cell line model, GE ethyl acetate extract (GEEA) (100 µg mL(-1)) treated group showed the maximum and significant (P < 0.05) growth inhibition at 48 h. The IC50 value was found to be 24.5 ± 19.1 µg mL(-1) at 48 h. Moreover, a low level of LDH release was observed at 48 h at various concentrations of (40, 60, 80 and 100 µg mL(-1)) GEEA extract-treated group compared to a control group. Changes in the cell morphology and echinoid spikes formation were observed at 48 h. Safety evaluation of GEEA in a non-cancerous liver cell line, PBMC and in Wistar rats positively revealed that the extract did not show any adverse toxic effects. The GEEA extract was partially purified by column chromatography and the active fraction was characterized through LC-MS analysis. Furthermore, HPLC and FT-IR analysis of the active fractions confirmed the presence of phytol, a diterpene compound with potent antiproliferative activity, which positively suggests that the red alga G. edulis contains a potent anticancer active principle.

A correlation of fecal volatiles and steroid hormone profiles with behavioral expression during estrous cycle of goat, Capra hircus.

Chemical signals (both volatile and non-volatile) form the major communication channels in animals. These signals are transferred mainly through excretory sources to facilitate inter-individual communication. In particular, the reproductive cycle of female mammals, including goats, exhibits significant changes in the constituents of their excretory products, and female mammals also express different behavioral patterns. We propose that feces is one of the important sources of chemo-signals in goats. However, the behavioral patterns and analysis of excretory sources based on chemical communication have not yet been studied in the Indian goat, Capra hircus. To validate our hypothesis, we analyzed the behavioral patterns and the volatiles and steroid hormone profiles in the feces samples of female goats during the estrous cycle. Here, we synchronized the estrous cycle in six female goats and obtained feces samples. The samples were extracted with dichloromethane and analyzed using gas chromatography-mass spectrometry. A portion of the sample was used for hormone assay to confirm the phases in the estrous cycle. Induction of she-goats into estrus was detected from the vaginal swelling, mucus discharge, restlessness, reduced milk secretion, bellowing, bleating, frequent urination, standing heat, allowing the male to mount, mounting on other females and teasing of males. The repeated male behaviors viz., flehmen, mounting, penile protrusion, body rubbing, dominance over other males and finally coitus with estrus female by male goats were observed. Analysis of volatiles revealed a total of twenty-four compounds combining all the phases in the estrous cycle. Among those, some of the volatile compounds and two antioxidants (ascorbic acid and vitamin E) were estrus-specific. Based on the fecal steroid analysis, higher level of estradiol during estrus and higher level of progesterone during post-estrus were observed. The behavioral patterns of female and male goats combined with qualitative differences in the volatile compounds and the two antioxidants rendered the estrus identifiable. Furthermore, the fecal steroid analysis also supported the detection of hormonal status during the estrous cycle. To the best of our knowledge, this is the first report correlating the behavior with volatiles and hormones in the feces samples from female Indian goats. It is concluded that the volatile pattern and hormone profile in feces, supported by specific behavioral patterns, should be considered a better modality of non-invasive estrus detection in goats.

Faecal chemical cues in water buffalo that facilitate estrus detection.

Chemo-signals are among the reliable non-invasive methods for estrus detection in mammals. Water buffalo is a silent heat animal and, hence, there is search for chemo-signals which would be effective non-invasive indicators of estrus state. We analyzed the faecal chemical cues during the estrous cycle in buffalo and to find the estrus-specific faecal volatile compounds adopting bull behavior assay. The faecal samples were collected at three phases of the estrous cycle (i.e., proestrus, estrus and postestrus) and subjected to gas chromatography-mass spectrometry analyses. We found 27 volatile compounds in the faeces of buffaloes, of which 4-methyl phenol (4mp) and trans-verbenol (tv) were found only in estrus faeces. The faecal samples of estrus buffaloes and the estrus-specific compound(s) (4mp+tv) at three different concentrations were tested for behavioral responses (flehmen and mounting behavior) in the bull. The bulls exhibited repeated flehmen when exposed to a combination of the two compounds (i.e., 4mp+tv) as compared to the individual compounds or raw faecal sample collected from buffalo when in estrus (P<0.05). However, higher number of mounting behavior was recorded when bulls were exposed to 4mp followed by a combination of the two compounds (4mp+tv) and trans-verbenol (P<0.05), in that order. By contrast, less number of mounting behavior was exhibited by bulls when exposed to the control sample (i.e., Hexadecanoic acid) (P<0.05). As inferred from the bull behavior assay, the present study suggests that the two compounds, 4 methyl phenol and trans-verbenol would be reliable indicators of estrus in buffaloes.