The chitin and chitosan biopolymers are extremely valuable because of their numerous industrial and pharmacological uses. Chitin and chitosan were extracted from the exoskeleton of Periplaneta americana (cockroaches) and termites using various acid and alkali techniques. The extraction process involves an initial demineralization step, during which integument dry powder was subjected to 500 mL (2.07 mol/L) of concentrated HCl at 100 degrees Celsius for 30 min, followed by meticulous rinsing with distilled water to restore the pH to its baseline. Deproteinization was conducted at 80 degrees Celsius using 500 mL (1 mol/L) of NaOH solution, which was repeated for 24 h. A total of 250 mL (0.06 mol/L) of NaOH was added at 100 degrees Celsius for 4 h to obtain chitosan, followed by extensive washing and subsequent drying. FTIR analysis was used to identify the functional groups in Periplaneta americana and termites. The crystallinity of these biopolymers, which have a face-centered cubic structure, was determined by X-ray diffraction analysis. This study assessed the analgesic properties of chitin and chitosan via an acetic-acid-induced writhing test in mice, revealing a significant reduction in writhing behavior following the chitin and chitosan extract. Notably, chitin exhibits the highest degree of analgesic activity compared to chitosan. Both chitin and chitosan show anti-inflammatory effects, with chitosan absorbing proton ions at sites of inflammation, while chitin effectively inhibits ear edema and elicits an analgesic response in mice. Furthermore, the present study revealed antipyretic activity, with termite chitin demonstrating the most significant effect at a concentration of 500 µL/mL, followed by chitosan and chitin at 100 µL/mL. These findings indicate the potential of using chitin and chitosan derived from termites and Periplaneta americana as natural anti-inflammatory compounds, implying prospective uses in anti-inflammatory, antipyretic, and analgesic capabilities.

Background and Aim: Trypanosomiasis, a parasitic infection caused by various Trypanosoma species, poses a significant threat to global livestock, affecting both human health and economic sectors. This study aimed to estimate the prevalence of trypanosomiasis in Southern Punjab, Pakistan, focusing on key ruminant species, including camels, cattle, buffaloes, goats, and sheep. Materials and Methods: A total of 240 blood samples, comprising 48 samples from each animal species (camel, cattle, buffaloes, goat, and sheep) were collected from three districts in Southern Punjab. The collected samples were subjected to thin smear microscopy, DNA extraction, and polymerase chain reaction (PCR) amplification. The molecular characterization was conducted using the TBR primer set, which targeted repeated satellite DNA regions and the cytochrome oxidase II gene of Trypanosoma brucei. Results: About 22.08% (53/240) of overall samples were positive for trypanosomiasis, with prevalence rates being 23.75% (19/80), 21.25% (17/80), and 21.75% (17/80) for districts Muzaffargarh, Lodhran, and Bahawalpur, respectively. 5.83% (14/240) of samples tested for T. brucei using PCR were positive in the districts of Muzaffargarh 7.50% (6/80), Lodhran 5.00% (4/80), and Bahawalpur 5.00% (4/80). Among the animals tested, camels had the highest positivity rate. The microscopic examination confirmed infection rates of 45.83% (22/48) for camels, 18.75% (9/48) for cattle, 8.33% (4/48) for buffaloes, 18.75% (9/48) for goats, and 18.75% (9/48) for sheep (p < 0.001). PCR results did not reveal substantial differences (p < 0.05) in prevalence: camels 12.50% (6/48), cattle 6.25% (3/48), buffaloes 0% (0/48), goats 8.33% (4/48), sheep 2.08% (1/48); while distinct disparities were detected district-wise: Muzaffargarh 23.75% (19/80), Lodhran 21.25% (17/80), and Bahawalpur 21.25% (17/80). The PCR results for these districts were insignificantly different: 7.50% (6/80), 5% (4/80), and 5% (4/80). The microscopic infection rate in camels from Bahawalpur was 56.30% (9/16). The microscopic analysis in Buffaloes reported a 6.30% (1/16) infection rate, but PCR results indicated no infections (0%) in any district. A significant difference (p < 0.001) in identifying Trypanosoma species was found between positively and negatively tested animals in both microscopic and PCR methods. Conclusion: This study emphasizes the necessity of regularly using PCR-based screening for its superior sensitivity and specificity over traditional microscopy. The varying occurrence of trypanosomiasis among districts reflects the intricate nature of this diseases epidemiology in the region. Reducing economic losses from trypanosomiasis in Southern Punjab, Pakistan, requires targeted interventions, such as vector control measures and farmer education. Keywords: domestic ruminants, microscopic examination, Pakistan, polymerase chain reaction detection, Trypanosoma brucei, trypanosomiasis.

Parasitic diseases, notably babesiosis, exert a substantial impact on the global cattle industry, posing challenges to commerce, economies, and human health. This study, conducted in Southern Punjab, Pakistan, aimed to assess the prevalence of Babesia spp. across various livestock species using microscopic and PCR methods. A total of 180 blood samples (60 from each district) were systematically collected from apparently healthy animals, with 36 samples obtained from each domestic animal species, including camel, cattle, buffalo, goat, and sheep, noting that 12 samples were collected from each district for each animal species. Overall prevalence was determined to be 32.8% (59/180), with varying rates among species: 25.0% in cattle, 41.66% in buffalo, 30.55% in goats, 33.3% in sheep, and 33.3% in camels. Microscopic examination revealed slightly varied infection rates among large and small domestic animals (22.2%), while PCR results indicated a 32.8% overall infection rate in both large and small domestic animals, with no statistical significance. District-wise analysis showed regional variations, with Muzaffargarh recording a prevalence rate of 23.33% through microscopic examination, while Lodhran and Bahawalpur recorded 21.67%. PCR results revealed higher rates (38.33%, 26.67%, and 33.33%, respectively), underlining the importance of employing PCR for accurate detection. Examining ruminant types, large ruminants exhibited a 32.4% infection rate, while small domestic animals showed 33.3%, with no significant difference (p=0.897). District-wise prevalence showcased significant variation, with Muzaffargarh demonstrating a 25% prevalence, Lodhran 22%, and Bahawalpur 22%, through microscopic examination. PCR results displayed 38.33%, 27%, and 33.3%, respectively, with no statistical significance. Detailed analysis of individual districts highlighted variations in infection rates among camels, cattle, buffalo, goats, and sheep. The binomial test indicated significant differences through microscopic analysis (P=0.011) but non-significant variations through PCR (P=0.065), emphasizing the precision of PCR. Regional variations in prevalence, notably with Punjab exhibiting the highest frequency (33.87%) and KPK the lowest (13.24%), suggest potential influences from varying veterinary practices and environmental factors. This study underscores the pivotal role of PCR alongside microscopy for accurate babesiosis diagnosis. These findings contribute to the broader understanding of babesiosis prevalence, emphasizing the necessity of advanced molecular techniques for informed control measures.

Background and Aim: Coccidiosis, caused by protozoan parasites of the genus Eimeria, is a significant concern in poultry farming, leading to substantial economic losses worldwide. In Pakistan, poultry is a major component of the agricultural sector, with both broiler and egg-laying chickens playing crucial roles in meeting the country’s protein needs. Despite the importance of the poultry industry, there is limited data on prevalence and species distribution of Eimeria in different types of chickens in District Swabi, Khyber Pakhtunkhwa, Pakistan. This study aims to estimate the prevalence and determine the distribution of Eimeria species in broiler and egg-laying chickens in this region. Materials and Methods: Nine hundred fecal samples were collected from broiler (380) and egg-laying domestic chickens (520) in District Swabi, Pakistan. Microscopic analysis was used to identify Eimeria parasites in all samples. After microscopic examination for positive identification, Eimeria species were determined using polymerase chain reaction (PCR) assays. Results: Microscopic examination identified Eimeria oocysts in 44.4% (400/900) of the samples. Eimeria parasite infection significantly varied based on chicken type, age, and gender (p < 0.05). The study found that broiler chickens (52.63%, 235/450), young chickens (4–6 weeks) (55.5%, 285/500), and females (52.2%, 200/380) were more infected with Eimeria spp. than egg-laying domestic chickens (38.5%, 200/520), adults (above 6 weeks) (28.8%), and males (36.7%, 165/450). PCR indicated a distribution rate of 42.5% (170/400) Eimeria tenella, 26.25% (105/400) Eimeria acervulina, 20% (80/400) Eimeria maxima, and 11.25% (45/400) Eimeria mitis. None of Eimeria necatrix, Eimeria brunetti, or Eimeria praecox was found in the study. Conclusion: This study underlines the essential requirement for targeted interventions due to the prevalence and predominance of E. tenella among identified Eimeria species. Future research should focus on refined sampling strategies and investigate the clinical significance of these parasites for effective disease management in the local poultry industry. Keywords: coccidiosis, domestic chickens, Eimeria species, microscopic and molecular identification, Pakistan, prevalence.

Limited research exists on the breeding ecology of the black francolin (Francolinus francolinus) in northern Pakistan. This study assessed egg dimensions, clutch size, hatching, fledging, and overall breeding success across different habitats and nests (n = 25) at Totali Game Reserve, Buner. Generalized linear models (GLMs) were used to analyze the effects of nest site characteristics and nest traits on breeding parameters. Egg dimensions were consistent across sites whereas bush nests had slightly wider eggs. The average clutch size was 5.9 ± 1.7 eggs, with an average of 4.8 ± 1.0 hatchlings per nest. A total of 111 chicks fledged, averaging 4.4 ± 1.0 per nest, yielding an overall breeding success rate of 75.5%. Nests containing six eggs had higher hatching success (76.6%). GLMs results showed a significant positive relationship between clutch size and hatchling, while nest site and traits had no significant effects. However, fledgling success was positively influenced by hatchling numbers, with nests in wetland habitats yielding significantly more fledglings (4.6 ± 0.9) than those from dryland habitats (4.0 ± 1.2). These findings suggest Black Francolins prefer nesting in wetland areas in bushes, likely due to better protection and favorable conditions.

The standard technologies and hazardous chemicals used in the synthesis of silver nanoparticles (Ag-NPs) can be harmful to the environment and as well as for human health. Sustainable and eco- friendly method of synthesizing nanoparticles are required. To synthesize silver nanoparticles (Ag-NPs), Grewia optiva leaf extract was used in the current research as a reducing agent. The leaves were thoroughly cleaned and dried before forming in an aqueous solution using a Jeldal apparatus for 20 minutes at 100 ºC. For the purpose, the extract and the silver nitrate (AgNO3) solution were thoroughly mixed. The method of synthesizing nanoparticles was optimized through the manipulation of multiple parameters, such as pH, temperature, and salt content. It was found that the ideal temperature for the synthesis of nanoparticles was 85 °C, the ideal pH range was 6-7, and the ideal salt concentration was 2 mL. Different techniques were used for characterization, including X-ray diffraction (XRD) to determine the crystalline structure of nanoparticles, Fourier-transform infrared spectroscopy (FT-IR) to identify chemical bonds and functional groups, using an Energy Dispersive X-ray Spectrometer (EDX) and transmission electron microscopy (TEM) to determine particle size in detail, and scanning electron microscopy (SEM) to analyze morphological aspects. The biosynthesized Ag-NPs showed potent antibacterial, phytotoxic, insecticidal, antipyretic, and analgesic properties in addition to efficient radical scavenging. The results showed that Ag-NPs made from G. optiva aqueous extract are appropriate for the production of novel medical and agricultural remedies. Since this study was carried out in a lab, more investigation is required to determine whether large-scale production is feasible and to look into possible uses of these Ag-NPs in the agricultural and biomedical sectors.