Muhammad Hafeez

Association of detoxification enzymes with butene-fipronil in larvae and adults of Drosophila melanogaster

Insecticide resistance is a major challenge in successful insect pest control as the insects have the ability to develop resistance to various widely used insecticides. Butene-fipronil is a novel compound with high toxicity to insects and less toxicity to the non-target organisms. In the present study, the effect of butene-fipronil alone and in combination with three enzyme inhibitors, piperonyl butoxide (PBO), diethyl maleate (DEM), and triphenyl phosphate (TPP), was carried out on larvae and adults of Drosophilia melanogaster. Our results indicated that the co-toxicity indices of butene-fipronil + PBO, butene-fipronil + TPP, and butene-fipronil + DEM mixtures were 437.3, 335.0, and 210.3, respectively, in the second-instar larvae, while 186.6, 256.2, and 238.5, respectively, in the adults, indicating synergistic effects. Interestingly, butene-fipronil increased the expression of CYP28A5 in the larvae; CYP9F2, CYP304A1, CYP28A5, and CYP318A1 in the female adults; and CYP303A1 and CYP28A5 in the male adults. Furthermore, high-level expression of Est-7 was observed in the female adults compared to larvae and male adults. Our results suggest that there is no difference in butene-fipronil metabolism in larvae and male and female adults of D. melanogaster.

Isolation and functional identification of three cuticle protein genes during metamorphosis of the beet armyworm, Spodoptera exigua

The beet armyworm, Spodoptera exigua (Hubner), is one of the major crop pests and is a target for current pest control approaches using insecticides. In this study three cuticular protein genes CPG316, CPG860 and CPG4855 have been cloned from 0u2009h pupal integument of S. exigua through race PCR Strategy. The deduced amino acid sequences were found to contain the RR-2 consensus region of other insect cuticular proteins and construct phylogenetic trees for each protein. Using quantitative RT-PCR, the developmental expression of the three genes through several larval and the early pupal stages was studied. All three genes contribute to the endocuticle although CPG316 may have a different role from the other two genes. All three newly isolated genes were analyzed and their functions were determined by using direct injection of the dsRNA into early 5th instar larvae. All genes are expressed in the larvae and early pupae but in different patterns. Furthermore, phenotypic results show that these genes have differing effects on the development of cuticle, its flexibility and a big role in metamorphosis in both larval and pupal stages.

Fitness parameters of Plutella xylostella (L.) (Lepidoptera; Plutellidae) at four constant temperatures by using age-stage, two-sex life tables

Different temperature zones have significant impact on the population dynamics of Plutella xylostella. Effective management of P. xylostella requires the knowledge of temperature tolerance by different life stages. In the current study, fitness parameters of diamondback moth were reported by using age-stage, two-sex life table traits at four constant temperatures (15, 20, 25 and 30u202f°C). The life cycle of P. xylostella was significantly longer at 15u202f°C. The 20u202f°C level of temperature was found optimal for fecundity, gross reproductive rate (51.74 offspring) and net reproductive rate (44.35 offspring per individual). The adult pre-oviposition period was statistically at par at all four level of temperatures. However, the survival was maximum at 20u202f°C as compared to other three temperature ranges. Based on the current study, it was concluded that temperature has a great role in population build-up of P. xylostella and effective management tactics should be applied to prevent significant damage to cabbage and other cruciferous crops when the temperature in the field is near 20u202f°C.

Gossypol-induced fitness gain and increased resistance to deltamethrin in beet armyworm, Spodoptera exigua (Hübner): Gossypol-induced fitness gain and increased resistance to deltamethrin in Spodoptera exigua (Hübner)

BACKGROUNDnIn plant-insect interactions, phytotoxins such as gossypol, exert a defensive role on behalf of the plant by interfering with the essential metabolic, biochemical and physiological pathways of herbivorous insects. The beet armyworm, Spodoptera exigua (Hübner), is a key pest for many important crops including a range of vegetables, ornamentals, and cotton. In this study, we investigated how resistance to deltamethrin relates to enzyme activity in gossypol-pretreated larvae of S. exigua.nnnRESULTSnFollowing selection with deltamethrin insecticides on gossypol-pretreated larvae for 10 generations, the Gos-SEL population developed a 113.29-fold resistance. Under the same conditions, the Delta-SEL selected population showed a 69.76-fold increase in resistance along with corresponding levels of xenobiotic defense enzyme activity. Similarly, the fecundity of the Delta-SEL population together with male and female longevity were found to be significantly lower when compared with the Gos-SEL population and the laboratory susceptible-strain group (SS-Strain). In addition, the activities of cytochrome P450s in S. exigua were significantly enhanced when the insects were fed on a deltamethrin and gossypol-pretreated diet compared with being fed on deltamethrin alone.nnnCONCLUSIONnThe reproductive capacity of S. exigua is significantly reduced in Delta-SEL and Gos-SEL populations compared with the control group (SS-Strain). Elevation of the major detoxification enzyme cytochrome P450 monooxygenase and esterase might have an important role in inducing tolerance to deltamethrin in gossypol-fed S. exigua populations. This study enhances our understanding of detoxification enzyme pathways for S. exigua gene expression and their role in responses to insecticides and plant secondary metabolites.

Gastric emptying scintigraphy in postprandial distress syndrome

Objective: To find out the pattern of gastric emptying scintigraphy (GES) in patients with post prandial distress syndrome (PDS). Methods: This study was carried out from January 2015 to July 2016 at Combined Military Hospital (CMH) Kharian and Nuclear Medical Centre (NMC) of Armed Forces Institute of Pathology (AFIP) Rawalpindi. Patients inclusion criteria were dyspepsia of post prandial distress type for more than six months duration. Patients with dyspepsia due to epigastric pain syndrome and other organic disorder were excluded. Upper gastrointestinal endoscopy was performed in all patients to rules out organic causes. Four-hour Gastric emptying scintigraphy was carried out at NMC, AFIP. Results were compiled and statistical assessment was done by utilizing SPSS IBM 22 version. Results: Thirty-eight patients were included in the study with age range from 15-72 years with mean age of 37.05±13.5 years. Males were 28(73.7%) and 10(26.7%) were female. Mean gastric retention with SD at one, two, three and four hours were 63 ± 19.04, 37± 20.62, 19±16.66 and 10±12.73 percent respectively. Early gastric emptying was in 3(7.89%) and delayed gastric emptying at two and four hours was seen in 4(10.52%) and 12(32%) respectively. Seventeen (44%) of the patients had normal gastric emptying despite the classical symptoms of PDS. Conclusion: Gastric dysmotility in GES seen in half of the patients points some additional mechanism as well like gastric accommodation or visceral hypersensitivity in the patients with PDS.

Microscopic cuticle structure comparison of pupal melanic and wild strain of Spodoptera exigua and their gene expression profiles in three time points

The beet armyworm, Spodoptera exigua (Hubner), is one of the major crop pests and is a target for current pest control approaches using insecticides. S. exigua melanic mutants (SEM) spontaneously occurred in the S. exigua wild type (SEW) strain and have been maintained under laboratory conditions on an artificial diet. Scanning electron microscopy showed that the inner cuticle of the SEM had a denser and less orderly structure. We investigated the cuticle protein genes using RNA-seq at three different developmental stages of both SEM and SEW. Comparison of cDNA libraries showed that 7257 CPs were significantly up-regulated and 664 genes were significantly downregulated in SEM at the developmental stage of 46-h in the fifth instar. In addition, 460 genes were significantly up-regulated and 439 genes were significantly down-regulated in the SEM at the development stage of 4-h before pupation. Moreover, 162 genes were significantly up-regulated and 293 genes were significantly downregulated in the SEM, just after pupation. Two genes CPR63 and CPR97 were identified from RNA sequences to verify the differentially expressed gene (DEG) results through quantitative real-time PCR (qRT-PCR). The results show that expression of both CPR63 and CPR97 structural cuticular proteins were significantly different between SEM and SEW. This functional analysis may help in understanding the role that these genes play in the cuticle pattern of the SEM.

Transcriptomic analysis of differentially expressed genes and related pathways in Harmonia axyridis after sulfoxaflor exposure

Harmonia axyridis (Pallas) is a major biological control agent that is indirectly threatened by sulfoxaflor insecticide applications targeting pests in agro-ecosystems. However, the genome-wide response of H. axyridis to sulfoxaflor remains largely unknown. Here, we used high-throughput RNA-seq technology to characterize the variation in the transcriptomic profile of second-instar H. axyridis larvae after exposure to sulfoxaflor. Two libraries were generated, with 50,702,976 and 47,005,096 clean reads for the sulfoxaflor treatment and control, respectively; these were grouped into 112,497 transcripts and 52,229 unigenes, of which 33,820 and 25,175 unigenes were respectively matched to proteins in the NCBI (Nr) and Swiss-Prot databases. From these unigenes, 794 significant differentially expressed genes (DEGs) consisting of 446 up- and 348 down-regulated genes, namely P450s, GSTs, USTs, ESTs, and others, were identified between the sulfoxaflor-treated and control groups. Furthermore, these DEGs were involved in several KEGG pathways including drug metabolism and metabolism of xenobiotic pathways. For transcriptome validation, 10 specifically expressed genes were selected and verified by quantitative real-time PCR analysis. Together, these results demonstrated that sulfoxaflor induces notable variation in the transcriptome profile and pathways of H. axyridis, which establishes a basis for further molecular investigations.

The trade-environment nexus in light of governance: a global potential

The theme of paper is to explore the trade-environment relationship and the role of institutions for 117 countries from global standpoint and five regions: Sub Saharan, European, Middle East and North Africa, Asia and Pacific, and Latin America and Caribbean, using the panel data span 2002–2014. By considering the endogeneity problem, to validate the nature of trade-environment nexuses, we applied the GMM first difference model in two steps. Likewise, Dumitrescu-Hurlin panel causality analysis is employed to affirm the causal relationship among the concern variables. The empirical findings of this study validate that the overall trade is significantly good to environment for sample countries (117) of the entire World, Europe, Asia, and Pacific regions. In further assessment, we incorporate interactive terms of institutions with trade, scale effect, and scale-technique effect. The estimated results confirm that institution is the persistent instrument for resolving the environmental problems. Furthermore, we find the evidence of inverted u shape EKC in overall selected sample of the World, Sub-Saharan, Europe, Asia, and Pacific regions. In contrast, there is no confirmation of inverted u shape EKC hypothesis in Middle East and North Africa regions. Similarly, no strong evidence of inverted u-shaped EKC hypothesis is observed in Latin-America and Caribbean region.

Sucralfate and Lidocain: Antacid 50:50 solution in Post Esophageal Variceal Band Ligation Pain.

Objective: To compare the effectiveness of pain relief of Sucralfate and lidocain antacid 50:50 solution in post esophageal variceal band ligation pain. Methods: All patients who had under gone Esophageal Variceal Band Ligation (EVBL) were included in the study. Patients un-willing to be included in the study or those who didn’t have post EVBL pain were excluded. Patients with post EVBL pains were divided into two groups: one group was given sucralfate and other was given lidocaine: antacid 50:50 solution. Both were inquired about the duration of the pain relief after the medication. The results were analyzed on SPSS 23. Independent samples T-test was performed to find out whether the difference in duration of pain relief was significantly different in the two groups Results: Out of 110 patients who have EVBL, 66(60.00%) had pain and 44(40.00%) were pain free. In the pain group 46 (69.7%) were given sucralfate and 20 (30.3%) were given lidocain: antacid 50:50 solution. Mean duration of pain relief in two groups was 2.78 (SD ± 2.096) and 2.5 days (SD ±. 0.76) respectively. Independent samples T-test results revealed that there was no statistically significant difference in the duration of pain relief between these two groups with p value 0.426. Conclusion: Both Sucralfate and Lidocain: antacid 50:50 solutions are effective in relieving the post EVBL pain. However, no statistically significant difference in duration of pain relief was detected in separate groups of patients treated with either treatment.

Ruptured hydatid cyst in biliary tract.

A lady aged 26 years reported with a 2 months history of fever, upper abdominal pain and weight loss. Her abdominal ultrasonographic scan revealed a complex cystic mass in left lobe of liver suggestive of hydatid cyst that was confirmed on magnetic resonance imaging of abdomen and magnetic resonance cholangiopancreatogram. With strong suspicion of a hydatid cyst, endoscopic retrograde cholangiogram was performed which confirmed the diagnosis. During the procedure, hydatid membranes protruding from the papilla were removed after sphincterotomy. She was put on albendazole 400 mg twice daily after the procedure and showed a remarkable clinical improvement.