Asad Ullah

Products derived from organisms play an important role in the discovery of new treatments of different chronic and debilitating diseases. Examining the components of annelids has provided a slew of biologically active substances over the years. Antifungal, antibacterial, anti-allergic, antioxidant, anti-inflammatory, and a variety of other properties medically useful properties were found in annelid components. This study investigated the antioxidant, anti-inflammatory, and anti-angiogenic properties of Pheretima posthuma an annelid Asian species. The earthworm's methanol and aqueous extract were tested after it was sacrificed (05 percent ethanol). The anti-angiogenic efficacy of Pheretima posthuma was investigated in-vivo utilizing a Chorio Allantoic Membrane (CAM) assay using distilled water extract., the antioxidant capacity of earthworm (Pheretima posthumous) was determined using methanol extract and in-vitro antioxidant tests of 2,2-Diphenyl-1-picrylhydrazyl (DPPH) and hydrogen per oxide (H₂O₂). A membrane stabilizing assay and a heat induce hemolysis assay utilizing distilled water extract were used to regulate the anti-inflammatory activity of earthworm hypotonicity (HRBCs). Protein denaturation was inhibited by 59.72 percent in the HRBC membrane stabilizing experiment at 80µg/ml. The aqueous extract of Pheretima postmortem exhibits the maximum inhibition at 80µg/ml, i.e., 62.16 percent, in the HRBC membrane stabilizing experiment. The extract was synthesized and tested using an antioxidant DPPH scavenging assay, it produced a great outcome at 40µg/ml, was 54.01 percent. The maximum H₂O₂ scavenging capacity of the Pheretima posthuma methanol extract was recorded 69.80 percent at 80µg/ml. The highest anti-angiogenic activity was detected at 80µg/ml, i.e., 62.16 percent using aqueous extract of Pheretima posthuma. In conclusion, the earthworm extract caused significant changes and has the capacity to influence angiogenesis-growth of new blood vessels; to prevent and treat neoplastic diseases in human beings.

| Toxoplasmosis is an infectious disease of protozoan origin infecting wide range of domesticated animals and poses significant threat to human population as well. Toxoplasma gondii is transmitted to ruminants through ingestion of oocytes-contaminated herbage. The purpose of the current study was to report prevalence and access the risk factors associated with toxoplasmosis. A total of 652 specimens of blood were taken from 9 regions situated within the jurisdiction of Punjab province, Pakistan. The collected samples were labeled in order to have randomization and to pull 384 samples out of 652 harvested samples. Survey Tool Box was used to have representative samples. Latex agglutination test (LAT) was performed on 384 the collected sera. Research data showed that there was high prevalence in sheep than in goats at 29.35% and 13.11%, respectively and there was high prevalence in Mundri breed and lowest in Thalli at 40% and 10.96%, respectively. The district wise data showed that Bhakkar has highest prevalence 31.30% and Khanewal showed lowest prevalence 11.40%. Data showed that Non-lactating animals showed high prevalence 35.29% and lactating animals have low prevalence 10.74%. Pregnant and non-pregnant animals have big differences of 10.68% and 38.66% prevalence, respectively. Prevalence rate was high by using the Bio-Scien kit at 29.5% and was low by using Atlas kit at 13.04%. Parity base prevalence was highest in animals more than 4th parity (22.65%) and lowest in animals 1-2 parity was 11.11%. Females showed more prevalence rate 31.53% as compared to male animals 8.02%. The results indicate that the disease is prevalent in the small ruminant population hosted in Punjab and more studies are needed to molecularly characterize T. gondii variants.

| Ticks (Acari: Ixodidae) are obligate ectoparasites of diverse hosts that affect livestock globally and are carriers of several bacterial, viral, and protozoan infections that affect both animals and human. Ticks (Class Arachnida) are ectoparasites of a wide variety of vertebrates, including livestock, and wild animals. Ticks are arachnids of veterinary and medical importance because they can transmit various diseases. Accurate identification of tick species is crucial for effective disease surveillance, prevention, and control strategies. Morphological identification and epidemiology based on the examination of key external features remain a fundamental and widely used approach for tick taxonomy. This research work provides a comprehensive overview of the morphological characteristics used in the identification of ticks, focusing on the main genera and species of medical and veterinary significance. The purpose of the current study was to determine the prevalence of tick infestation, epidemiological characteristics, associated risk factors, and investigation of a variety of hard tick infestation in small ruminant populations in District Mardan, Buner, and Dir Lower, Khyber Pakhtunkhwa, Pakistan. A total of 5000 goats and sheep of various ages, breeds, sexes, and locations were inspected and examined for tick infestation. A total of 1774 goats and 1044 sheep were found to be tick infested, and the percentage prevalence was 59.37 for goats and 51.45 for sheep were recorded. Our studies showed tick diversity, infestation rate, and numerous factors (season, age, and gender of host) influencing tick infestation rate in goats and sheep population in District Mardan, Buner, and Dir lower, Khyber Pakhtunkhwa. Among the identified species Haemaphysalis and Rhiphicephalus were the most prevalent genera. To build a precise nationwide model of tick infestation, more study is required to confirm our findings in the area and to validate our findings in other locations of Pakistan. It is possible to boost domestic animal productivity while reducing the spread of deadly tick-borne illnesses in society by educating farmers about ticks and providing nearby veterinary services. A large-scale study is needed to explore the hard tick’s diversity across the country and awareness is needed to minimize the risk of infection, especially among farmers and farms owners.

The black soldier fly larvae Hermetia illucens L. (Diptera: Stratiomyidae) are an ideal insect to produce compost and animal feed. The goal of this study was to use Kitchen waste 50 grams, Cow dung 50 grams, and Rotten fruits 50 grams attractants to gather wild H. illucens and their egg clutches from their natural microhabitat in dumping sites. The fly was trapped in a 1×1×1 feet trap net. The results indicate that humidity 42.5±50.5%, F = 1.754, P = .233 and temperature 27.975±37.05ºC, F = 1.126, P = .353) during the capture of H. illucens and egg clutch in a natural environment. The life cycle has included seven instars, a duration of 39 days, and the longest time period of the instar 6th recorded. We have optimized the temperature range to 24-30ºC and humidity to 60-95% in indoor rearing of the H. illucens. We recorded the varying ingredient contents level, highest Moisture at 38.5%, Crude protein at 55.13%, Crude fiber at 28.18%, Crude fat at 28.18%, and Total Ash at 15.5%. The H. illucens indoor farming is recommended for quality of protein production under a controlled environment which will boost the Pakistani economy and reduce threats to aquatic fauna.

| Nile Tilapia ( Oreochromis niloticus ) is the most common marketable fish species in biofloc throughout the world. It has higher growth rate and is reared in limited resources when compared to other aquaculture species. This research work identified the requirements needed for installation of biofloc setup in winter session; different physico-chemical parameters and optimized the floc density for growth of Nile Tilapia ( O. niloticus ) in the study area. A total of three tanks were taken and the floc concentration was 121.071g in tank first, 131.16g in second and 135.19g in tank third. Total fingerlings (750) stocked were divided in three tanks (dimensions of length was 15feet, height=4.5feet, water level=3.2 feet) with water volume of 17000 liters in each tank. Formulated feed was used with 30% and 40% crude protein (CP) and buffalo meat. Water temperature was maintained at 31 o C using heater (500Watts) with thermostat. The results were recorded with significant value in the growth of fish during 4 th to 9 th weeks. The values of fish weight were recorded higher (P<0.05) in group 3 than the group 1 and group 2. The floc concentration, feed and physico-chemical parameters have positive effects on biofloc system at winter session for quality production of Nile Tilapia ( O. niloticus ) production.

| Pesticides are extensively used throughout the world in industrial, health sector for agriculture and domestic purposes. Despite beneficial effect, these pesticides when released into the environment lead to induced toxicity in a large number of non-target organisms. Some of these are non-biodegradable and are persistently present in environment leading to environmental pollution. These chemicals eventually reach aquatic bodies and cause various histopathological, hematological, bio-chemical and enzymatic alterations in the bodies of aquatic organisms, especially fish, leading to huge economic loss. Consumption of the affected fishes also poses a serious health threat to humans. Pakistan being an agricultural country uses a variety of pesticides to protect its crops. The use of pesticides has substantially increased in Pakistan over the last decades, which when reach the water bodies, adversely affect the rich biodiversity found in aquatic systems of Pakistan. This review discusses research over past decades regarding toxic effects of pesticides induced in edible freshwater fishes of Pakistan and future considerations.

| This study was conducted to detect the effect of inorganic selenium on the liver enzymes, kidney function and changes in biochemical parameters of broiler chicken under dexamethasone (DEX) induced stress. The day old, two hundred (n=200) chicks were bought from commercial hatchery and randomly divided into five groups with eight replicas(n=5) in each group. Broiler chickens were fed with starter and finishing commercial corn based basal diet (BD) with different doses of inorganic Selenium (Se) powder. Group-A was kept negative control provided with BD twice a day. Group-B was positive control group fed with 15mg DEX/kg twice/day. The Group-C was fed with BD+0.2mgSe+15mg DEX/kg. Similarly, Group-D was fed with BD+0.3mgSe+15mgDEX. Group-E was fed with BD+0.4mgSe+15mg DEX/kg. Two chicks were randomly slaughtered from each replicate and blood glucose level, kidney function and liver enzyme were examined. After slaughtering, the collected blood was centrifuged for 15 minutes at 1000 rpm for serum analysis and anticoagulant added tube for whole blood. The serum was stored at -20°C until further analysis. Biochemical parameters were measured through commercial kits. The result of the current study showed that the values of Alanine transaminase (ALT), Aspartate aminotransferase (AST) and Alkaline phosphatase (ALP) were higher significantly (P≤0.05) in negative control group (B) as compared to the group C, D and E. The level of uric acid, creatinine and blood urea were significantly higher (P≤0.05) in group A (negative control) (P≤0.05) as compared to the group B (positive control) C, D, and E. The blood cholesterol level and low-density lipoprotein (LDL) in group B (positive control) was significantly higher (P≤0.00) than the group A (negative control) and Se supplemented group C, D and E.

Aedes aegypti are slender, fragile, long-legged mosquito, which are distributed worldwide. In general, it is a terrestrial and nocturnal species that act as pollinator for few plant species as well as it is a hematophagous insect and act as main vector for many arboviruses (dengue, yellow fever, etc.). In many cities of Pakistan, dengue fever is becoming more prevalent as an infectious disease. as there is currently no vaccine for the dengue virus and control of the disease depends on vector control. The present study aims to investigate morphological and molecular identification of Aedes aegypti in district Buner. A total of 365 samples of mosquitoes were randomly collected from six tehsils of the district Buner. Before examination, all the collected samples were preserved in labelled Eppendorf tubes containing silica gel and transported to Veterinary Research & Disease Investigation Center (VR&DIC), Balogram Swat, Khyber Pakhtunkhwa, Pakistan for further laboratory analysis. Morphological identification was performed with the help of stereomicroscope using taxonomical keys. To increase identification accuracy, each sample was reconfirmed by PCR using mitochondrial Cytochrome oxidase first gene (COI). The amplified PCR product was sequenced, and the obtained sequences were trimmed by Bio Edit and BLAST in NCBI. After BLAST, phylogenetic tree was constructed by using MEGA-X. In phylogenetic tree, the sequence (COI) genes of Aedes aegypti species as previously reported from Sri Lanka and India. Furthermore, the results of this study will generate basic information about mosquitos spp; and it will help to understand the epidemiology of Aedes aegypti disease in Khyber Pakhtunkhwa, especially in district Buner, Khyber Pakhtunkhwa, Pakistan.

The present study was conducted on bovine mastitis in three tehsils of district Karak, Khyber Pakhtunkhwa Pakistan. A total of 124 cow milk samples were collected randomly and were screened for presence of subclinical mastitis through surf field mastitis test (SFMT) and polymerase chain reaction (PCR)and microbiological procedures to isolate Staphylococcus aureus (S. aureus). Furthermore, the disc diffusion technique was applied, and the isolate of positive S. aureus was phenotypically assessed for methicillin-resistant S. aureus (MRSA) and vancomycin-resistant S. aureus (VRSA). The overall results showed that out of 124 milk samples, 16.93% (21/124) were found positive for subclinical mastitis on the Surf Field Mastitis Test (SFMT). Further, amongst these 21 positive samples, 8 (38.09%) samples were found positive for S. aureus when swabbed on Mannitol Salt Agar (MSA). Through PCR, the resistant genes were amplified and identified. In each of these 8 positive samples, genes were examined for mecA, tetK, etA, etB, fbe, embP, and IS256. Additionally, good results were found for mecA, fbe, embP, and IS256. The S. aureus isolates displayed 100% resistance to Erythromycin,followed by Cefoxitin, Cefradin and Augmentin (75%); Ciprofloxacin(62.5 %);Gentamicin (50%);Clindamycin (50%) followed by Amikacin and Tigecycline who showed 25% resistance. Vancomycin and Linezolid were the only antibiotics that shown zero resistance and were discovered to be susceptible to study isolates.

The Black solider fly Hermetia illucens, larvae meal (BSFLM) is a good source of protein alternative to fish meal (FM). The quality of fish fishmeal has been challenged economically in the aquaculture feed industry. The current research was designed to identify the varying ingredient content levels of black soldier fly products (BSFP), recorded highest crude protein 42% and moisture 28.5% in mature fly, crude fiber 20.5%, and total Ash 14.5% in BSFL shell, crude fat 25.5%, in the pupa. The Black soldier fly larvae meal was tested 6-week experiments on Ctenopharyngodon idella 60 fingerlings (initial average length 1.58inch, 3.4gm weight and final body length and weight (4.38±0.10a, 15.00±0.00 a). Respectively, Channa marulius fingerlings initial average length is 6.52cm, weight 3.52gm and gain final body length and weight (8.50±0.18 a, 6.00±0.16 a). The Sixty fingerlings of each fish were divided into three groups: commercial and BSFLM feed. The feeds conversion ratio (FCR) of C. idella (2.564) and C. marulius (2.155) was calculated with the commercial base diet. The BSFLM (FCR) value of C. idella (1.4368) and C. marulius (2.106). The growth rate has a significant difference (P <0.05) and the strongest correlation recorded, temperature, ammonia, total dissolved solids, and water pH to affect the fish growth. No adverse effects were noted from the utilization of BSFLM, the formula was recommended to be used for C. idella and C. marulius fish. We concluded from our study that the BSFLM supplementation shown promising biological effects and cost-effective alternative protein and energy sources for C. idella and C. marulius fish in term of their growth traits.