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This study investigates the molecular prevalence and phylogenetic characteristics of two prominent blood-borne pathogens, Toxoplasma gondii (T. gondii) and Plasmodium spp., in common quails (Coturnix coturnix) sampled from both wild (N = 236) and farmed (N = 197) populations across four districts (Layyah, Dera Ghazi Khan, Lahore, and Multan) in Punjab, Pakistan, during the hunting seasons from 2021 to 2023. Additionally, the impact of these pathogens on the complete blood count (CBC) of the hosts is examined. Out of 433 quails tested, 25 (5.8%) exhibited amplification of the internal transcribed spacer (ITS-1) gene for T. gondii, while 15 (3.5%) showed amplification of the Cytochrome b gene for Plasmodium spp. A risk factor analysis indicated that the prevalence of both pathogens was not confined to specific sampling sites or bird sexes (P > 0.05). District-wise analysis highlighted that hens were more susceptible to both T. gondii and Plasmodium spp. infections than cocks. Wild quails exhibited a higher susceptibility to T. gondii compared to farmed birds. Significant CBC variations were recorded in infected birds as compared to uninfected ones. BLAST analysis of generated sequences has confirmed the identity of recovered PCR amplicons as T. gondii and Plasmodium relictum. Phylogenetic analysis revealed that Pakistani isolates clustered with those reported from various countries globally. This study provides the first documentation of T. gondii and Plasmodium sp. infections in Pakistani quails, underscoring the need for detailed investigations across different regions to enhance our understanding of infection rates and the zoonotic potential of these parasites. [ABSTRACT FROM AUTHOR]

Bovine mastitis is a widespread and costly disease that affects dairy farming globally, characterized by mammary gland inflammation. Bovine intramammary gland infection has been associated with more than 135 different pathogens of which Staphylococcus aureus is the main etiology of sub-clinical mastitis (SCM). The current study was designed to investigate the prevalence, antibiotic resistance pattern, and the presence of antibiotic resistance genes (mecA, tetK, aacA-aphD and blaZ) in S. aureus isolated from the raw milk of cows with subclinical mastitis. A total of 543 milk samples were collected from lactating cows such as Holstein Friesian (n = 79), Sahiwal (n = 175), Cholistani (n = 107), and Red Sindhi (n = 182) from different dairy farms in Pakistan. From the milk samples microscopic slides were prepared and the somatic cell count was assessed to find SCM. To isolate and identify S. aureus, milk was streaked on mannitol salt agar (MSA) plates. Further confirmation was done based on biochemical assays, including gram staining (+ coccus), catalase test (+), and coagulase test (+). All the biochemically confirmed S. aureus isolates were molecularly identified using the thermonuclease (nuc) gene. The antibiotic resistance pattern of all the S. aureus isolates was evaluated through the disc diffusion method. Out of 543 milk samples, 310 (57.09%) were positive for SCM. Among the SCM-positive samples, S. aureus was detected in 30.32% (94/310) samples. Out of 94 isolates, 47 (50%) were determined to be multidrug resistant (MDR). Among these MDR isolates, 11 exhibited resistance to Cefoxitin, and hence were classified as methicillin-resistant Staphylococcus aureus (MRSA). The S. aureus isolates showed the highest resistance to Lincomycin (84.04%) followed by Ampicillin (45.74%), while the least resistance was shown to Sulfamethoxazole/Trimethoprim (3.19%) and Gentamycin (6.38%). Polymerase chain reaction (PCR) analysis revealed that 55.31% of the isolates carried blaZ gene, 46.80% carried tetK gene, 17.02% harbored the mecA gene, whereas, aacA-aphD gene was found in 13.82% samples. Our findings revealed a significant level of contamination of milk with S. aureus and half (50%) of the isolates were MDR. The isolated S. aureus harbored various antibiotic resistance genes responsible for the absorbed phenotypic resistance. The alarmingly high prevalence of MDR S. aureus isolates and MRSA strains in these cases possess a serious risk to public health, emphasizes the urgent need to address this issue to protect both human and animal health in Pakistan. [ABSTRACT FROM AUTHOR]

Introduction: The present study evaluated the effect of preslaughter stress intensities on the behavioral, physiological, blood biochemicals, and hormonal responses in goats. Methods: Twenty-seven intact male goats (Ardi breed, 10 months of age, 27 kg liveweight) were divided into three treatment groups viz., the control (C) group, ear pulling (EP) group, and leg pulling (LP) group. Various behavioral, physiological, blood biochemical and hormonal responses were recorded before and after handling. Results and Discussion: The EP and LP goats had a higher frequency and intensity of vocalization as compared to control goats. The preslaughter handling stress intensities had a significant effect on the before and after handling values of heart rate, respiration rate, rectal temperature, and ear temperature. Further, among groups, the glucose value increased significantly upon preslaughter handling as compared to the baseline value. The LP goats had significantly higher after-handling value for lactate dehydrogenase (LDH) as compared to the before-handling value. The catecholamines (adrenaline and noradrenaline) and β-endorphin concentrations increased significantly upon preslaughter handling. The higher physiological, behavioral, blood biochemical, and hormonal response indicated higher preslaughter stress in EP and LP goats. Both levels of intensity revealed unfavorable responses in goats that may adversely affect animal welfare and meat quality. Thus, to ensure minimal adverse effects on behavior, physiology, blood enzymes, and hormonal responses, it is recommended to follow animal welfare principles when implementing preslaughter handling practices. [ABSTRACT FROM AUTHOR]

Importance: The creation of robust maternal-embryonic interactions and implantation models is important for comprehending the early stages of embryonic development and reproductive disorders. Traditional two-dimensional (2D) cell culture systems often fail to accurately mimic the highly complex in vivo conditions. The employment of three-dimensional (3D) organoids has emerged as a promising strategy to overcome these limitations in recent years. The advancements in the field of organoid technology have opened new avenues for studying the physiology and diseases affecting female reproductive tract. Observations: This review summarizes the current strategies and advancements in the field of 3D organoids to establish maternal-embryonic interaction and implantation models for use in research and personalized medicine in assisted reproductive technology. The concepts of endometrial organoids, menstrual blood flow organoids, placental trophoblast organoids, stem cell-derived blastoids, and in vitro-generated embryo models are discussed in detail. We show the incorportaion of organoid systems and microfluidic technology to enhance tissue performance and precise management of the cellular surroundings. Conclusions and Relevance: This review provides insights into the future direction of modeling maternal-embryonic interaction research and its combination with other powerful technologies to interfere with this dialogue either by promoting or hindering it for improving fertility or methods for contraception, respectively. The merging of organoid systems with microfluidics facilitates the creation of sophisticated and functional organoid models, enhancing insights into organ development, disease mechanisms, and personalized medical investigations. [ABSTRACT FROM AUTHOR]

This experiment aimed to ascertain whether adding chitosan-oligosaccharides (COS) to the diet will affect Mandarah laying hens' productivity and reproductive efficiency. At the age of 34 weeks, 120 laying hens and 12 cocks of the Mandarah chicken strain were separated into four groups, each consisting of 30 hens and three cocks. The 1st group served as a control group, receiving only a basic diet. The 2nd, 3rd, and 4th experimental groups received 0.1, 0.2, and 0.5 g/kg of chitosan-oligosaccharides as an addition to the base diet, respectively. According to the findings of this study, the majority of the analyzed attributes differed significantly between treatment groups. Comparing the COS-treated birds to the control birds, the COS-treated birds significantly increased egg production, improved feed conversion, Haugh unit score, shell thickness, fertility, hatchability, ovary weight, and the number of yellow ovarian follicles as well as blood estradiol-17β (E2) and testosterone concentrations. Furthermore, compared to the other treated groups, introducing COS at a level of 0.1 g/kg diet resulted in the best laying rate, egg mass, and feed conversion. The highest percentage of fertility, hatchability of hatching eggs, ovary weight, number of yellow ovarian follicles, and serum E2 concentration was also found in birds fed COS at a dose of 0.2 g/kg feed. In addition, compared to the control diet, COS treatment at 0.1, 0.2, and 0.5 g/kg enhanced relative economic efficiency by 464.43, 457.41, and 352.75%, respectively. It is concluded that chitosan improves economic efficiency, reproductive performance, productivity in laying hens, and overall health when used at 0.1 g/kg diet. [ABSTRACT FROM AUTHOR]

Metabolic syndrome is one of the most frequent diseases with the augmentation of the rapid changes in societies' lifestyles. On the other hand, various medicinal plants have recently been extensively used for plentiful therapeutics. Among these, curcumin and turmeric are the main components derived from the rhizome of plants, with significant effects on preventing metabolic syndromes. Due to their antioxidant and anti-inflammatory activities, the biological activities of turmeric and its bioactive element, curcumin, have antidiabetic properties via augmenting insulin release and anti-hyperlipidemia effects via enhancing fatty acids uptake, anti-obesity properties via declining lipogenesis, and antihypertensive properties via augmenting nitric oxide synthesis. Moreover, the dietary presence of turmeric and its bioactive element, curcumin, significantly affected poultry productivity, health and welfare. According to human and animal trials, turmeric or curcumin has important values as dietary supplementation and complementary therapy in some metabolic syndromes. This scenario highlights on the potential effects of curcumin (turmeric) on poultry health and its role in complementary therapy in metabolic syndrome. Additional high-quality clinical trial investigations are required to establish the clinical efficiency of the curcumin complement confidently. [ABSTRACT FROM AUTHOR]

Background Cutaneous leishmaniasis (CL), an emerging vector-borne ailment in Khyber Pakhtunkhwa (KPK), Pakistan, exhibits diverse spread patterns and outbreaks. Methods To comprehend its epidemiology and identify parasite species, we conducted an active survey on suspected CL cases (n=8845) in KPK. Results Microscopy and internal transcribed spacer-1 PCR–restriction fragment length polymorphism (RFLP) molecular techniques detected Leishmania spp. in blood samples. Phylogenetic analysis gauged genetic affinities with other areas. District Bannu displayed the highest CL impact (14.58%), while Swat had the lowest impact (4.33%) among cases. Annual blood examination rate, parasite incidence and slide positivity rate were 4.96 per 1000 people, 0.0233 and 0.047%, respectively. CL infections were prevalent in 1- to 20-y-olds, with males (57.17%) more vulnerable than females (42.82%). Single lesions occurred in 43.73% of patients, while 31.2% people had two lesions, 17.31% had three lesions and 7.74% had more than three lesions. Most had sand-fly exposure but lacked preventive measures like repellents and bed nets. Leishmania tropica was confirmed via RFLP analysis in amplified samples. Phylogenetic analysis unveiled genetic parallels between L. tropica of KPK and isolates from China, Iran, Afghanistan, India, Syria and Morocco. Conclusions Urgent comprehensive control measures are imperative. Early detection, targeted interventions and raising awareness of CL and sand-fly vectors are vital for reducing the disease's impact. International collaboration and monitoring are crucial to tackle Leishmania spp.'s genetic diversity and curtail its cross-border spread. [ABSTRACT FROM AUTHOR]

This study aimed to determine the effects of Coenzyme Q10 (CoQ10) in the freezing medium on functional and oxidative stress parameters and in vitro fertilization (IVF) rate of buffalo sperm. Collected samples were relocated to the laboratory for initial evaluation, gentle dilution in extenders, cooling (4°C, 2 h), equilibration (4°C, 4 h), packaging (straws, 0.5 mL), programmable freezing, and thawing (37°C, 30 s). Statistical analysis depicted that adding CoQ10 (100 μM) in a freezing medium caused a significant augmentation in total motility (%), average path, and straight‐line velocities (μm/sec) of buffalo sperm than control. Adding CoQ10 (100 μM) improved sperm progressive motility, rapid velocity, and functional parameters (%) compared to the control and 10 μM of CoQ10. Moreover, CoQ10 in a freezing medium caused a significant augmentation in seminal plasma catalase (U/mL) and glutathione reductase (GSH; nmol/109) at 100 μM than control and other treatments. CoQ10 inclusion (100 μM) ameliorates seminal plasma superoxide dismutase (U/mL), glutathione‐S‐transferase (GST; nmol/mL/min) fructose (μg/mL), and ATP (nmol/million) than control. Furthermore, CoQ10 at 100 μM improved seminal plasma glutathione peroxidase (μM) levels than control, 10 μM, and 20 μM. Lastly, hydrogen peroxide (H2O2; nM) production was significantly lower at 100 μM than at control and 10 μM. CoQ10 (100 μM) caused a significant augmentation in the un‐capacitated pattern followed by a reduction in the capacitated pattern, and apoptosis‐like changes (%) than control, and other treatments, whereas viability was increased than control and other treatments. CoQ10 (100 μM) significantly improved the IVF rate in comparison with control, CoQ10 at 10 μM, and 20 μM groups. In conclusion, the addition of CoQ10 (100 μM) in the freezing medium can improve the quality and in vitro fertility of post‐thawed buffalo semen via its antioxidative effect. Further studies are needed to evaluate the effect of CoQ10 on the in vivo fertility of buffalo bull semen. [ABSTRACT FROM AUTHOR]

Background: A hydroponic feeding system enables more effective utilization of the entire plant than typical grazing, which only consumes the plant's shoot. Aim: This study evaluated the effects of feeding maize hydroponic fodder on growth performance, nitrogen balance, nutrient digestibility, hematology, and blood metabolites of buffalo calves. Methods: Twelve water buffalo calves, weighing an average of 112 + 1.18 kg and between 8 and 10 months old, were divided into three treatments, each with four calves. Each group received one of the treatment diets: T1: the basal diet (BD) at 100%; T2: the BD plus hydroponic feed meal (HFM) at 80%: 20%; and T3: the BD plus HFM at 60%: 40%. For 100 days, each animal was fed ad libitum; the first ten days were used for nutritional adaptation, and the final ten days were used for collection. In addition to their BD, each animal received 200 g/day of a normal concentrate mixture to meet their maintenance needs. The BD included Green Hay (Lucerne) 80% and Wheat straw 20%. Each animal's daily feed consumption was noted. Calves were weighed biweekly to track growth. Upon completion of the experiment, blood samples were obtained. Results: The amount of dry matter (DM) consumed by ruminants fed diets, including hydroponic fodder, was considerably higher (p < 0.05). Similar trends were seen in crude protein (CP), acid detergent fiber, and neutral detergent fiber intake. Ingesting of CP was highest in animals fed T3. Animals fed diets comprising BD 60% + HFM 40% had the highest levels of DM and CP digestibility. Animals fed the T3 diet (BD 60% + HFM 40%) showed the best feed conversion values (p < 0.05). Blood metabolites like blood urea nitrogen, creatinine, and glucose showed non-significant variations in all experimental animals. In hematology, a similar trend was seen. Conclusion: Therefore, it can be said that supplementing the diet with more HFM helped growing buffalo calves gain weight, have a lower feed consumption ratio, and digest their food more efficiently. [ABSTRACT FROM AUTHOR]

Background: Feed additives that increase nutrient availability in feeds have gained a lot of interest. Aim: An experiment was conducted to determine whether amylase, protease, and their combined supplementation affected broiler performance. Methods: Two hundred eighty broiler chicks were selected and distributed randomly into 28 replicate pens with four treatment groups and seven replicates under a completely randomized design. A total of four diets were developed, having 0, 100, 100, and 100 + 100 g of control (AP0), amylase (A1), protease (P1), and amylase + protease (AP1)/ton of feed, respectively. Four replicates of each treatment were fed each diet. Each diet was randomly allotted to each group. Ad-libitum feeding was provided to the birds. The feeding program had starter and finisher diets. Upon completion of the experiment, three birds from each pen were slaughtered to analyze the carcass characteristics and organ weight. Results: Differences were insignificant between 100 g/ton of amylase supplementation and FI, body weight gain (BWG), or feed conversion ratio (FCR) (p > 0.05). Supplementation with 10 0g/ton of protease did not significantly affect FI, BWG, and FCR (p > 0.05). Similarly, 100 + 100 g/ton of amylase + protease addition had no significant effect on FI, BWG, and FCR (p > 0.05). None of the treatments significantly affected carcass weight, abdominal fat percentage, dressing percentage, drumstick, wings, breast, and thigh weights (p > 0.05). In addition, there were no significant effects (p > 0.05) on the weight of the heart, liver, gizzard, and spleen. Conclusion: In conclusion, amylase, protease, and their combined supplementation at a rate of 100 g/ton of feed did not influence BWG, FI, FCR, carcass characteristics, or organ weight. [ABSTRACT FROM AUTHOR]