Bisphenol-A (BPA) is a xenoestrogen that mimics the action of natural estrogen, estradiol-17 beta, and can disrupt the endocrine system. Present study was designed to investigate histopathological effects of BPA on some vital organs of major carp, Catla catla. Fish were exposed to various sub-lethal concentrations (1 to 4 ppm) of BPA in a semi-static system for 15 days. After the stipulated time, liver, kidneys and gills were dissected out and fixed immediately in 10 % buffered formalin. After embedding in paraffin wax, 5 micron thick sections were cut and stained with H & E and PAS stains. In liver, BPA caused central vein congestion, inflammation, edema, degeneration and necrosis of hepatocytes. Kidney anomalies included obliteration of bowmen's space, shrinkage and degeneration of tubules and glomerulus. Gills responded to BPA stress by hyperplasia of mucous cells, clubbing and degeneration of secondary-lamellae. The present study revealed that BPA causes degenerative changes in various vital organs of C. catla and severity of histological changes were dose-related.

The aims of the study were to determine immune function circulating leukocytes using an ex vivowhole blood stimulation assay (WBA) with lipopolysaccharide (LPS) and assess gene expression profiles by RNA sequencing in the transition period of dairy cows. The WBA was performed on whole blood of 6 Holstein multiparous cows at −20, −3, 3, 7 d from parturition (DFP) using 0, 0.01 and 5 µg LPS/mL. The plasma collected after stimulation was used to analyze IL-1β and IL-6 concentration via ELISA. The data were analyzed as a factorial design with repeated measures, using PROC MIXED in SAS. At the same day of the WBA test, RNA was isolated from whole blood and sequenced on a Hiseq1000 (Illumina, USA). Differential gene expression analysis was conducted with the edgeR package, and a general linear model was applied considering −20 DFP as the baseline. A threshold of 1.5-fold change and P < 0.05 were used to define differentially expressed genes (DEG), which were subsequently analyzed through the Dynamic Impact Approach (DIA) using the Kyoto Encyclopedia of Genes and Genomes (KEGG) database. The IL-1b and IL-6 released after LPS stimulation was higher at −3 DFP (P < 0.05) in comparison with −20 DFP. After calving, the response of IL-1β to stimulation of LPS decreased markedly, while the IL-6 response was unchanged up to 3 DFP (P < 0.01 vs. -20 DFP) and then declined at 7 DFP. The most-impacted and activated KEGG pathways highlighted by the DIA analysis at −3 vs. −20 DFP were: PPAR signaling, adipocytokine signaling, hematopoietic cell lineage, ECM-receptor interaction and phagosome. After calving (3 and 7 DFP) the impact and activation of the above listed pathways was strongly increased, but there also was a strong inhibition of arachidonic acid metabolism (in particular enzymes regulating leukotrienes synthesis) as well as glycine, serine and threonine metabolism. Overall, the WBA and transcriptomic data confirmed changes in immune-competence of the circulating leukocytes around calving and, in particular, indicate mainly an increase of their activity and function. These data support the idea that the dairy cow's immune system is dysfunctional but not immunosuppressed around calving.

The transition period is known to be the most critical phase in the life of high yielding dairy cow. Changes in the immune functions have been observed during the transition period which may account for the onset of clinical and subclinical (e.g. inflammatory response) problems at calving or at the beginning of lactation however this relationship has not yet been adequately investigated. Thus, to establish the potential of the periparturient dairy cow's immune system to respond to stimuli, two challenges [an ex vivo whole blood stimulation assay (WBA) with lipopolysaccharides and a carrageenan skin test (CST)] were performed in addition to characterizing the metabolic and inflammatory profile. The WBA was performed using 0, 0.01 and 5 μg LPS/mL on whole blood and CST was administered by subcutaneous injection of 0.7 mL solution containing 4.2mg of carrageenan to the shoulder region of the cows. These tests were performed on 10 Holstein-Friesian cows at -45 ± 2, -20 ± 2, -3, 3, 7, 28 ± 2 days from parturition (DFP). Cows were also monitored for health status, body condition score, milk yield. The results demonstrate a higher production of IL-1β and IL-6 from leukocytes after LPS stimulation around calving (from -3 to 3 DFP) compared to -45 DFP (P < 0.05). Moreover, IL-6 (but not IL-1β) was able to reach close to the maximum response at the lower stimulus intensity (0.01 μg LPS/mL), maintaining a higher response over a longer time in early lactation. The release of higher levels of IL-6 in the transition period, with low LPS dose, suggests its crucial role in the regulation of inflammatory response around calving. The response of cows to CST decreased a few days before calving (-3 DFP) compared with response at -45 and 28 DFP (P<0.05), and remained low in the first week of lactation. This result suggests the reduction of the functionality of some vascular factors, which decreases diapedesis. Overall, the WBA and CST tests confirm changes in immunocompetence around calving. These tests are able to better describe the changes of the innate immune response at a local and systemic level, mainly when combined with conventional metabolic and inflammatory indices.

The dietary preferences and paleoecology of Giraffokeryx punjabiensis Pilgrim, 1910, an extinct giraffid species from Chinji Formation of Potwar Siwaliks of Pakistan, have been investigated by incorporating mesowear-I, mesowear- II and hypsodonty methods. The mesowear patterns of Giraffokeryx punjabiensis are very much consistent with the browsers as well as seasonal mixed feeders whereas hypsodonty index (1.31 +/- 0.06) categorizes it within the browsers. Comparison of the results from these research tools suggest that the extinct giraffid reflects diverse dietary spectra ranging from browsers to seasonal mixed feeders and shows no affinities with grazers. The coexistenceof G. punjabiensis with its mammalian paleocommunity reveals the persistence of mosaics of diverse habitats ranging from tropical evergreen forest to subtropical ones, closed seasonal woodlands to wooded savannas.

Acute acidosis was induced in sheep, and gastrointestinal permeability was assessed by using lactulose as a permeability marker. Metabolism was evaluated by monitoring blood metabolites. Four rams (72.5 ± 4.6 kg BW) were used in a 2 × 2 changeover design experiment. The experimental period lasted 96 h from -24 to 72 h. After 24 h of fasting (from -24 to 0 h) for both controls and acidosis-induced rams (ACID), 0.5 kg of wheat flour was orally dosed at 0 and 12 h of the experimental period to ACID, while the basal diet (grass hay, ad libitum) was restored to control. At 24 h, a lactulose solution (30 g of lactulose in 200 mL of water) was orally administered. Blood samples were collected at -24, 0, 24, 48, and 72 h of the experimental periods for the analysis of metabolic profiles and during the 10 h after lactulose dosage to monitor lactulose changes in blood. In addition, rumen and fecal samples were collected at 24 h of the experimental period. The acidotic challenge markedly reduced (P < 0.01) rumen pH and VFA but increased rumen d- and l-lactic acid (P < 0.01). Concurrently, a decrease of fecal pH and VFA occurred in ACID (P < 0.01), together with an abrupt increase (P < 0.01) of lactate and fecal alkaline phosphatase. Blood lactulose was significantly increased in ACID peaking 2 h after lactulose dosage. Blood glucose, β-hydroxybutyrate, Ca, K, Mg, and alkaline phosphatase showed a significant reduction (P < 0.05) at 24 h, whereas urea and NEFA declined (P < 0.05) from 48 to 72 h. A strong inflammatory acute phase response with oxidative stress in ACID group was observed from 24 to 72 h; higher values of haptoglobin (P < 0.01) were measured from 24 to 72 h and of ceruloplasmin from 48 (P < 0.05) to 72 h (P < 0.01). Among the negative acute phase reactants, plasma albumin, cholesterol, paraoxonase, and Zn concentration also decreased (P < 0.05) in ACID at different time points between 24 and 72 h after acidotic challenge start. A rise (P < 0.05) of reactive oxygen metabolites and a drop of vitamin E (P < 0.01) between 24 and 72 h were indicative of oxidative stress in ACID. The perturbation of these blood metabolites suggests that acute acidosis was effectively induced by our model. The increase of lactulose in blood in ACID indicates that gastrointestinal permeability for the marker increased and the large increment after 2 h from dosage suggests that most of the passage occurred through the rumen or abomasal walls.

The resistance/susceptible status of Aedes aegypti (field strain) from slum area of Misri Shah Lahore, Pakistan was determined during May 2009 to January 2011. Two insecticides i.e. deltamethrin 1. 5% EC and cypermethrin 10% EC which were sprayed in various localities of Lahore from the last ten years we re used to evaluate resistance/ susceptible status of field collected adult females of Aedes aegypti by CDC bottle bioassays. The resistance level was expressed as resistance ratio (RR) of lethal time at 50% death determined using field collected and susceptible strain s. Current study indicated RR LT50 1.95 and 1.47 against deltamethrin and cypermethrin in adult females of Aedes aegypti respectively. In addition, the tested population was less resistant against cypermethrin as compared to deltamethrin. In comparison of percent mortalities, there was no significant difference between the two insecticides against adult fema les.

Residual effect of Bacillus thuringiensis var. israelensis (Bti) Technical powder, WDG (water Dispersible Granules) and a Bacillus sphaericus (Bsph) was evaluated against laboratory-reared Anopheles stephensi and field collected Culex quinquefasciatus late third instars from Lahore, Pakistan. In general residual activity of both strains in seven different concentrations (100, 10, 1, 0.1, 0.01, 0.001 and 0.0001 ppm) for laboratory reared and in semi-field bioassays varied in both species of mosquito larvae. Bti TP residual activity varies from 51 days to 24 hours against laboratory reared A. stephensi larvae using maximum concentrations 100 ppm to minimum concentration 0.0001 ppm (fresh stock replacement every 24 hours). Whereas, Bsph TP indicated maximum residual effect for 18 days and minimum for 48 hours against the same species. Percent mortality was significantly higher (P=0.000) for A. stephensi against 100 ppm as compared to Bsph. However, the same effect (P=0.054) was observed for field collected Culex quinquefasciatus against 100 ppm Bsph. In conclusion field collected Culex quinquefasciatus larvae were more susceptible and have prolonged residual effect as compared to laboratory reared A. stephensi against Bsph while Bti have effect vice versa. Residual effect for field evaluation of Bti WDG under low treatment (0.2 mg/litre) against Culex quinquefasciatus lasted 14 days indicating more efficient for field bioassays as compared to laboratory.

In the current study, the status of resistance in Aedes aegypti was evaluated against pyrethroids insecticide. The resistance of field collected populations against Bifenthrin 10% EC was compared with the susceptible population. A range of different concentrations (40, 20,10,5 µg/ml) of Bifenthrin was used on susceptible laboratory reared population (reference population) to find the diagnostic dose which is 20 µg /ml. The diagnostic dose was used in two field collected populations i.e.Government Islamia College for Women Cooper Road, Lahore (GICW) and Government College University, Lahore (GCU) during July to September, 2010. Adult female mosquitoes were exposed to CDC bottle bioassays in two field collected populations i.e. GICW & GCU for the evaluation of resistance. Resistance level was expressed as resistance ratio (RR) of lethal time for 50% death determined in field collected and susceptible strain. Results of bioassays indicated that Aedes aegypti field population from GICW Lahore was resistant to Bifenthrin since 100% mortality occurred post 40 minutes exposure as compared to GCU where the same mortality occurred in 30 minutes post exposure. Resistance ratio (RR) also indicated that the population from GICW cooper road Lahore was found resistant (RR LT50 = 1.97 & LT95 = 1.5). Whereas, the field collected population of Aedes aegypti from GCU was found susceptible (RR LT50 = 1.003 & LT95 = 1.06) as compared to

In the current study, resistance status of Aedes aegypti was evaluated against pyrethroid insecticide. The resistance in field collected population against Deltamethrin 2.5% EC was compared with susceptible (laboratory reared) population. Field population was collected from two localities of Lahore i.e. Government College University and Government Islamia College for Women Cooper Road, Lahore during July- September, 2010. CDC Bottle Bioassays were carried out on adult female mosquitoes (susceptible) in order to determine the diagnostic dose. A range of concentrations (10, 5, 2.5 and 1.25 µg/ml) of deltamethrin was used for 60 minutes exposure. Diagnostic dose 1.25 ug/ml was found in post 30 minutes exposure. The same concentration caused 100% mortality of field collected populations from GCU and Govt. Islamia College for Women in 30 and 40 minutes exposure respectively. Resistance level was expressed as resistance ratio (RR) of lethal time for 50% death determined in field collected and susceptible strain. Results of bioassays indicated that Ae. aegypti field collected population from Government Islamia College for Women Cooper Road, Lahore was resistant to Deltamethrin at RRLT50 =1.3 and RRLT95=1.37 as compared to laboratory reared susceptible population. However, field population of Ae. aegypti from Government College University, Lahore was found susceptible at RRLT50=1.03 and RRLT95=1.0.

Of the seven genes encoding insulin-like peptides (ILPs) in the mosquito, Anopheles gambiae, four are arrayed proximally as duplicate pairs on chromosome three. Amino acid substitutions encoded in the duplicate genes occur in the C peptide and not the B and A peptides. Except for one duplicated gene, sequence-specific transcripts for all other AgamILPs were obtained from female mosquitoes. Transcript expression of each AgamILP was determined by RT-PCR in the head, thorax, and abdomen of all life stages and both sexes of this mosquito. Two AgamILPs were ubiquitously expressed, suggesting a growth factor function, whereas the other AgamILPs were expressed primarily in heads, as confirmed by the immunostaining of ILPs in the neurosecretory cells of female brains, thus indicating a hormonal function.

Midgut proteases contribute to the success or failure of Plasmodium infection of the mosquito. This paper examines the reciprocal effect of Plasmodium yoelii nigeriensis on midgut trypsin, chymotrypsin, aminopeptidase and carboxypeptidase in the mosquito Anopheles stephensi. The total protein ingested and the rate of protein digestion were unaffected by the parasite, but more protein was ingested at the first than the second bloodmeal. All peptidases were unaffected by the presence of the parasite during the first gonotrophic cycle, when ookinetes were penetrating the midgut. In the second gonotrophic cycle, trypsin and chymotrypsin were unaffected by growing oocysts, but aminopeptidase activity was reduced in the midguts of infected mosquitoes. Chymotrypsin activity was depressed and aminopeptidase activity elevated during the second gonotrophic cycle. Plasmodium infection has a negligible effect on bloodmeal digestion and does not limit the availability of the protein for egg production. The significance of changes in aminopeptidase activity when oocysts are present is discussed.

Our previous studies demonstrated a significant reduction in the egg production and survival of Anopeles stephensi Liston infected with Plasmodium yoelii nigeriensis Killick-Kendrick. We investigated the physiological mechanism underlying the malaria-induced curtailment of reproductive fitness. Polyclonal antibodies were raised against An. stephensi vitellin (Vn) and used in an enzyme immunoassay to quantify ovarian Vn and hemolymph vitellogenin (Vg) at 8, 12, 16, 20, 24, and 48 h postblood feeding in infected and noninfected mosquitoes. Initially, the concentration of Vg in the hemolymph and the accumulation of Vn in the ovaries of infected females were identical to females fed on noninfected mice, indicating that the synthesis of yolk protein by the fat bodies was initiated normally. However, there was a significant reduction in ovarian Vn at 24 and 48 h postblood feeding and a significant accumulation of Vg at 20 and 24 h postblood feeding in the hemolymph of infected mosquitoes. This increase in concentration of Vg in the hemolymph just before Vn reduction in the ovaries of infected mosquitoes indicated that synthesis at the fat body level may not be affected, but that sequestration of Vg by ovarian follicles of malaria-infected mosquitoes may be affected before resorbing follicles lose protein.

Infection with the rodent malarial parasite Plasmodium yoelii nigeriensis caused a significant reduction in the reproductive fitness (number of eggs produced and proportion of eggs hatched) of two different generations of Anopheles stephensi. Overall fertility (number of larvae produced) was reduced by 38.3% in the generation containing smaller mosquitoes (with a wing length of 3.2 +/- 0.1 mm) with relatively larger parasite burdens, and by 48.81% in the generation containing larger mosquitoes (with a wing length of 3.4 +/- 0.1 mm). The contribution that reduction in egg production and egg hatching made to overall reduction in reproductive fitness differed in each experiment. No significant difference was observed in the egg size (length from tip to tip and breadth at the centre) of control and infected mosquitoes in another generation of infected An. stephensi.