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The Chukar Partridge (Alectoris chukar) is a wild bird of the Phasianidae family. In Pakistan, it occurs in a wide range of rugged, sloping and dry areas, rising to the interior of the Himalayas, the western Himalayas and plains, the higher piedmont valleys on the dry slopes of the Baluchistan provinces and Punjab and Sindh provinces. It is the “national bird” of Pakistan, however, little is known about its ecology and reproduction in the northern part of the country. This study investigated the existence, distribution and population of birds in the Bajaur region of Khyber Pakhtunkhwa (KP) Province from December 2019 to August 2020. Usually in the early morning (5 to 78 AM) and evening (4 to 9 pm). The areas occupied by Alectoris chukar mainly include Mulberry, Ficus banyan, Gum arabic, Arabian acacia, Barberes lycium, Dilberia sisso, Melia azedarach, Ailanthus, Alalthus altissima, Zanthoxylum alatum, Olea europaea, Olea europaea, Ingres, Celtis eriocarpa and Pinus wallichiana. Habitat destruction (overgrazing, mowing and landslides) caused by hunting, shooting, capture, explosives, excavation, and road construction is the main threat to existing wildlife (including Alectoris chukar).

Obtaining oocytes from the adult female zebrafish (Danio rerio) ovary has enormous importance in the studies of developmental biology, toxicology, and genetics. It is vital to establish a simple and effective approach to ensure the quantity and quality of oocytes, which will enable the success of follow-up experimental investigation finally. Usually, oocytes are separated with mechanical or enzymatic methods, however, little studies have been done with concerns about the comparative effects. The present study separated zebrafish oocytes of Stage III with five frequently used methods, including stripping, pipetting, hyaluronidase (1.6 mg/ml), collagenase (0.4 mg/ml), and trypsin (0.1%). The cell viability, oxidative stress, mitogen-activated protein kinase (MAPK) protein phosphorylation, and apoptosis levels were selected as main biomarkers to evaluate the oocytes health status. The results showed that both trypsin and hyaluronidase isolation significantly upregulated germinal vesicle breakdown (GVBD) rates and downregulated p38 MAPK activity simultaneously. GVBD rates and survival rates were decreased notably in oocytes separated by the collagenase method. Above results indicate that zebrafish oocytes in vitro are sensitive to enzymatic treatments and the enzymatic isolation is not the suitable mean for collecting zebrafish oocytes although it is time-saving. The mechanical strategy of pipetting remarkably increased the reactive oxygen species and malondialdehyde level in isolated oocytes. Interestingly, oocytes separated with stripping show less physiological and biochemical damages. Therefore, stripping isolation is comparatively recommended as the optimum method for separating and collecting numerous intact and healthy zebrafish oocytes in vitro for the subsequent developmental research.

Cyanobacterial blooms and their associated toxins pose a great threat to human beings. The situation is even worse for those whose drinking water source is a cyanotoxin-polluted water body. Therefore, efficient and safe treatments urgently need to be developed. The present study verified the application of vitamin C on the inhibition of toxic Microcystis aeruginosa. Our results showed that vitamin C drove the Fenton reaction and significantly sterilized cultures of M. aeruginosa. The algicidal activity of vitamin C was dependent on its involvement in iron (Fe) metabolism. Vitamin C enhanced iron absorption leading to high ferrous ion levels. The ferrous ion increased production of reactive oxygen species (ROS) by Fenton reaction, which play a crucial role in the killing process. Interestingly, vitamin C also dramatically decreased the release of microcystins. This study highlights the possible benefits of using a vitamin C-induced Fenton reaction to remove M. aeruginosa and microcystins from drinking water sources.

We compared the bait consumption by rodents inhabiting poultry farms of Rawalpindi-Islamabad from i) wooden double opening bait station ii) PVC pipe bait station iii) cardboard bait station iv) earthen bowl bait station (3.5 cm deep and 17 cm diameter) made as safe as possible from non-target animals. The consumption of bait from the four different types of bait placement differed significantly. The amount of bait eaten from wooden double opening bait station was significantly greater than from the other three bait station designs. Least bait was eaten from the cardboard bait stations. Wooden double opening bait stations are essential tools for the application of rodenticides on poultry farms.

Locally available and palatable food grains viz. millet (whole), wheat (cracked), maize (cracked) and rice (broken) were tested for their preference to rodents. Results of no-choice and multiple choice feeding tests indicated that millet was the most preferred food. Similarly locally available taste additives namely peanut cracked (5%), yeast (2%), fish meal (5%), egg shell powder (5%), carbon disulphide (CS2) 30 ppm and jaggery (gur/sugar 5%) were offered mixed in millet-wheat (50:50 by wt.) bait. The results of paired choice and multiple choice feeding tests revealed that consumption of bait with added egg shell was significantly higher (p > 0.05) than bait with the other five additives. Grain bait with egg shell was then tested for its effectiveness as a carrier for the rodenticides zinc phosphide (2%) and coumatetralyl (0.0375%). The reduction in rodent activity after zinc phosphide and coumatetralyl treatment was 70% and 82%, respectively.

While microcystins (MCs) have been reported to exert reproductive toxicity on fish with a sex-dependent effect, the underlying mechanism has been rarely investigated. In the present study, zebrafish were exposed to 1, 5 and 20 μg/L MC-LR for 30 d. The gonad-somatic index declined in all treated males. 17β-estradiol (E2), testosterone (T), 11-keto testosterone (11-KT) and follicle-stimulating hormone (FSH) levels increased in serum from all treated females, while T, FSH and luteinizing hormone (LH) levels changed in all treated males. Histomorphological observation showed that MC-LR exposure evidently retarded oogenesis and spermatogenesis. Transcriptional changes of 22 genes of the hypothalamic-pituitary-gonad (HPG) axis exhibited sex-specific responses and the relationship between gene transcriptions and gametogenesis was evaluated by principle component analysis (PCA). Major contributors to PC1 (gnrh2, gnrhr3, ar, lhr, hmgra, hmgrb and cyp19a) were positively correlated with the number of post-vitellogenic oocytes, while PC1 (gnrh2, lhβ, erβ, fshr, cyp11a and 17βhsd) were positively correlated with the number of spermatozoa. The protein levels of 17βHSD and CYP19a were affected in both females and males. In conclusion, this study first investigated the sex-dependent effects of microcystins on fish reproduction and revealed some important molecular biomarkers related to gametogenesis in zebrafish suffered from MC-LR.

Cyanobacterial blooms caused by water eutrophication have become a worldwide problem. Microcystins (MCs), especially microcystin-LR (MC-LR), released during cyanobacterial blooms exert great toxicity on fish and even lead to massive death. The present study mainly investigated the pathological damage and immune response of spleen, gut and gill in zebrafish exposed to MC-LR. Fish were exposed to 0, 1, 5 and 20 μg/L of MC-LR for 30 d. In zebrafish exposed to 5 and 20 μg/L MC-LR, edematous mitochondria, deformation of the nucleus and compaction of chromatin were observed in lymphocyte of spleen; frayed gut villi, exfoliation of epithelial cells and widespread cell lyses were observed in intestines; hyperemia in gill lamellae, epithelial tissue edema and uplift and lamellar fusion were observed in gill. Varied changed gene expression was observed in spleen, intestine and gill of zebrafish. The transcriptional levels of IFN-1 and IL-8 in spleen significantly up-regulated in 20 μg/L group, and the transcription of IL-1β and TNFα in spleen increased in 1 μg/L MC-LR treated fish. In addition, the mRNA levels of IFN-1, IL-1β, IL-8, TGF-β and TNF-α dramatically increased in intestine and gill in all MC-LR treated groups. The present studies indicated that MC-LR exposure caused marked pathological damage, however, fish could adjust actively the expression of innate immune-related genes to resist the tissue damage. Our findings provided strong evidence of the recovery potential of fish exposed to microcystins.