University of Agriculture Faisalabad

The present study was conducted to evaluate the quality of milk sold at Tandojam, Pakistan. A total of 125 milk samples (25 samples from each source) collected from five milk marketing agencies viz direct seller (DS), milk collection center(MCC), milk vendor shop (MVS), hotel (HT) and buffalo dairy farm (DF), which served as control. Acidity of milk obtained from DS, MCC, MVS and HT averaged 0.13, 0.15, 0.12 and 0.13, respectively compared to 0.14 for DF milk. The pH values of milk from MVS (6.54) and HT (6.53) were significantly different (P 0.05) to that of DF milk i.e. 6.65. Viscosity, specific gravity and freezing point of milk procured from DS (1.48, 1.026 and –0.460, respectively), MCC (1.58, 1.026 and –0.470, respectively), MVS (1.34, 1.026 and – 0.440, respectively) and HT (1.46, 1.027 and –0.480, respectively) were significantly (P<0.001) lower than DF milk (1.86, 1.031 and -0.551, respectively). Chemical quality of milk procured from DS, MCC, MVS and HT compared to DF milk (control) averaged 13.45, 14.18, 13.19 and 14.06% vs. 16.30% for TS content, 8.25, 8.81, 8.06 and 8.51% vs. 9.79% for SNF content, 5.20, 5.41, 5.13 and 5.54% vs. 6.51% for fat content, 3.85, 3.96, 3.91 and 4.23% vs. 4.35% for protein content, 2.70, 2.77, 2.56 and 3.20% vs. 3.56% for casein content, 3.65, 4.03, 3.34 and 3.52% vs. 4.53% for lactose content and 0.75, 0.78, 0.74 and 0.76% vs. 0.91% for ash content, respectively. All the attributes of chemical quality of milk supplied through four agencies were significantly lower (P<0.05) than DF milk.

The increase in resistance pattern of Staphylococcus aureus (S. aureus) has been an emerging threat in therapeutic areas of the dairy industry throughout the globe. The current study was conducted in bovines of district Faisalabad, Pakistan to investigate the phenotypic prevalence of methicillin-resistant (MRSA) and vancomycin-resistant S. aureus (VRSA) in milk samples positive for subclinical mastitis. The study further aimed to assess the associated risk factors and antimicrobial susceptibility pattern against MRSA and VRSA isolates. A total of 385 milk samples (n=193 cattle; n=192 buffalo) collected and screened for subclinical mastitis by surf field mastitis test (SFMT) were further subjected to standard microbiological techniques for the isolation of S. aureus. The positive isolates of S. aureus were phenotypically evaluated for MRSA and VRSA by the disc diffusion method. The study results revealed that out of 385 milk samples, 45.97% (177/385) samples were found positive for subclinical mastitis on SFMT while 37.14% (143/385) samples were confirmed for the presence of S. aureus. Out of these S. aureus isolates, MRSA and VRSA were confirmed in 17.48% (25/143) and 12.58% (18/143) samples respectively. The in-vitro trials of various antibiotics for MRSA and VRSA isolates showed 100% resistance towards Cefoxitin followed by 50% towards Gentamicin, Tylosin, and Trimethoprim + Sulfamethoxazole then 25% to Oxytetracycline, and Fusidic acid while ciprofloxacin, moxifloxacin, and linezolid were found sensitive against study isolates. The public health importance of S. aureus and emerging resistance against antibiotics like methicillin and vancomycin demands regular monitoring of effective use of antimicrobial agents against the isolates of VRSA and MRSA.

Anticoccidial drug resistance has been increased due to which alternative protocols are required to control avian coccidiosis.In current study, Trachyspermum ammi (Ajwain) was supplemented in crude form in feed (3%) of broiler chickens which were given infection with sporulated oocysts of different Eimeria species.Toltrazuril ® anticoccidial drug was administered in control group.Anticoccidial activity of T. ammi was evaluated on the basis of different parameters such as feed conversion ratio, mortality rate, intestinal lesion, fecal scores, oocyst scores and organ weight.Data on hematological parameters and serum chemistry was also collected.Dietary supplementation of T. ammi reduced Eimeria infection in terms of better feed conversion ratio, lower lesion and fecal scores in broiler chickens (P>0.05).Supplementation of T. ammi also improved hematology and serum chemistry of infected chickens.It was concluded from the experiment that T. ammi can serve as alternative candidate against poultry coccidiosis however, further in vivo studies are needed to formulate its dose against coccidiosis.

Bovine babesiosis is a vector-borne parasitic disease of cattle that causes significant economic losses worldwide.Disease control is largely hampered by factors such as tick resistance and the lack of an effective vaccine.Other preventive methods, such as chemoprophylaxis, have high costs and resistance issues.Thus, to address this research gap, anti-idiotype antibodies against Babesia were prepared, and their immunomodulatory potential was compared with chemoprophylaxis.The current study was designed to analyze the immunomodulatory potential of Montanideadjuvanted anti-idiotype antibodies against experimental bovine babesiosis infection.Blood samples from PCR-confirmed Babesia (B.) bigemina and B. bovis-infected cattle were taken, and those parasitized erythrocytes were subjected to gamma-ray (γrays) irradiation.The irradiated, parasitized erythrocytes were inoculated into experimental goats to induce idiotype antibodies (IgG) against B. bigemina and B. bovis.The F(ab')2 fragments of these antibodies were injected into experimental layer hens to produce anti-idiotype antibodies (IgY).The anti-idiotype antibodies were subsequently evaluated for immunogenicity in experimental mice and immunomodulatory potential in calves.In experimental mice, group M Big showed mean antibody titers ranging from 76% to 83%, while M Bov showed titers of 74% to 82%.In experimental calves, the ELISA results showed that group C calves elicited a greater immune response at 21 days post-infection (DPI) for B. bigemina (optical density (OD) value=1.709)and B. bovis (OD value=1.698)as compared to calves of group D, which showed mean OD values for B. bigemina and B. bovis as 1.344 and 1.264, respectively.The results confirmed that anti-idiotype antibodies are an effective candidate for immunomodulation against bovine babesiosis.

The increasing population has led to the rising demand of poultry meat and eggs due to its exceptional nutritional value.To meet these environmental demands, studies are being conducted on innovative approaches to enhance the sustainability of poultry feed.Research have depicted alternative feed formulations that are low-cost yet high in nutrition.Fruits processing industries produce plethora of fruits waste every year.The utilization of agro-industrial (fruit) waste is considered as a potential source of low cost, highly nutritious poultry feed.In current research study fruits (apple, mulberry, apricot and pomegranate) waste (peel, pomace and leaves), rich in dietary fiber, protein, vitamins, minerals, amino acids, antioxidants, bioactive phytochemicals, tocopherols, flavonoids, polyphenols, phyto-sterols and essential fatty acids were focused to enrich poultry feed.Study results showed that with increasing concentration of fruits waste from T0 (Controlled diet) to T4 (87.5% commercial diet and 12.5% supplementation) in poultry feed, the phenolic content and antioxidants potential was increased (TPC: To = 0.25 g GAE/g → T4 =0.67 mg GAE/g, DPPH: T0 =1.64 → T4= 2.17, OH: T0 =1.84 →T4 =2.35).Mineral profiling of broiler feed showed a significant increase in potassium, calcium, magnesium, and phosphorus by the addition of agricultural wastes.On 42 nd day of study, experimental birds were evaluated for oxidative stability, fatty acid profile, cholesterol, HDL, LDL and triglycerides and found significant positive impact on poultry bird and its meat.Thus, fruit waste can be utilized as feed source of poultry animals to improve health and to reduce waste burden.

The emergence of triple-reassortant H3N3 avian influenza viruses (AIVs) poses a threat to poultry and public health.This study characterized three novel H3N3 isolates (SQ2049, HN0120, and GT2125) from chickens in China, evaluating their evolution, pathogenicity, and vaccine efficacy.Phylogenetic analysis revealed these isolates originated from H10N3 and H3N8 lineages, carrying an H9N2-derived internal gene backbone.Although identified as low pathogenic AIVs (IVPI=0), they exhibited distinct replicative fitness.GT2125 showed reduced efficiency in embryos, and carried unique PB2 substitutions (V598I and K702).Pathogenicity trials in SPF chickens demonstrated that SQ2049 achieves efficient horizontal transmission in SPF chickens without clinical signs.Notably, cloacal viral shedding persisted for 11 days, whereas oral viral shedding only lasted 2 to 3 day, with mild histopathological lesions observed in the trachea and immune organs.Importantly, the inactivated SQ2049 vaccine elicited high cross-reactive antibody titers (HI≥8log2) and provided 100% protection in SPF chickens.Mouse infection studies identified two HA substitutions (G174E and N209D) after viral replication in mice.These two mutations have previously been associated with mammalian adaptation and enhanced binding to human-type α-2,6 receptors, though their functional effects in the H3N3 context were not evaluated in this study.These findings expand our understanding of H3N3 AIV evolutionary dynamics and highlight the importance of surveillance for reassortant influenza viruses.The demonstrated vaccine efficacy provides a foundation for future H3N3 prevention and control strategies.

Diabetic cardiomyopathy (DCM) involves myocardial structural and functional remodeling, in which mitochondrial dysfunction and oxidative stress play central roles.Carnitine palmitoyltransferase 1A (CPT1A) may participate in diabetic cardiac injury, but its underlying mechanism remains unclear.A type 2 diabetes mouse model was established using a high-fat diet and low-dose streptozotocin, while H9c2 cardiomyocytes were exposed to high glucose and palmitic acid.CPT1A was overexpressed using AAV9 in mice and a lentiviral vector in cells.Cardiac injury, lipid metabolism, oxidative stress, and mitochondrial function were assessed through biochemical, histological, RT-qPCR and Western blot analyses.The regulatory relationship between FOXA1 and CPT1A was examined using gainand loss-of-function approaches.CPT1A expression was reduced in diabetic myocardial tissues and high-glucose/palmitate-treated H9c2 cells.CPT1A overexpression attenuated cardiac injury, partially improved FBG, LDL, TC and TG levels, reduced cardiomyocyte hypertrophy and fibrosis-related gene expression, restored mitochondrial respiratory chain activity and ATP production, and alleviated oxidative stress.FOXA1 was identified as a potential upstream regulator of CPT1A.FOXA1 overexpression reduced cardiomyocyte injury, whereas FOXA1 knockdown aggravated mitochondrial dysfunction, which was partially reversed by CPT1A restoration.CPT1A alleviates diabetic cardiac injury by preserving mitochondrial function and limiting oxidative stress.The FOXA1-CPT1A axis may represent a potential therapeutic target for DCM.

Zoonotic food-borne trematodes are transmitted to humans via aquatic vegetables, fish, crabs and crayfish as part of the freshwater ecosystem.Though both parasites may be delivered by the same food vehicle, the infective metacercariae are found in different biological food vehicles making global comparisons and food-safety surveillance difficult.A systematic evidence synthesis was performed as an integrative approach and included: indexed biomedical literature, systematic reviews, meta-analyses, primary prevalence surveys and authoritative WHO and EFSA reports.Evidence was grouped into three prespecified pathways as follows: (1) Fasciola spp. on aquatic vegetation; (2) Clonorchis spp., Opisthorchis spp.and Paragonimus spp. on freshwater crustaceans.Pooled estimates were not recalculated, but primary surveys were compared based on food vehicle, host species, production system and diagnostic method.Quantitative evidence was most well developed for fish-borne trematodes.The prevalence of C. sinensis metacercariae in fish was estimated from 71 studies and found to be 30.5%,with figures of 35.1, 29.7 and 8.4% in China, Korea and Vietnam, respectively.Overall prevalence of zoonotic trematodes in individual Vietnamese and Lao surveys ranged from about 40 to 69%.The prevalence of these second intermediate hosts in China was 52.07%for P. westermani and 31.10% for P. skrjabini.Fasciola metacercariae of edible aquatic plants were poorly documented: only F. hepatica DNA was found in 2 of 500 samples of water-dropwort in Korea and low but persistent metacercarial contamination of natural beds of watercress was found in France over a long period.The presence of zoonotic metacercariae in freshwater food systems is widespread; however, there is unevenness among the evidence both taxonomically and geographically.Estimates based on fish, crustacean and plant should not be combined into a single prevalence estimate without stratification.For credible global surveillance, food-unit sampling, species confirmation, viability assessment and metadata for the supply chain must be harmonized.

The unique ecological environment of the Qinghai-Xizang Plateau has a significant impact on the genetic differentiation of parasitic organisms.However, the information on the mitochondrial genome of ectoparasites on goats in this region is extremely limited.Flea samples were collected from the body surface of grazing goats and the genome was annotated using MITOS2.A phylogenetic tree was constructed using the maximum likelihood method, and a comparative analysis was conducted with known species of the flea family.A total of two complete mitochondrial genome sequences (G2 and G3) were obtained, with lengths of 16,238 bp and 16,274 bp, respectively.They contain 13 protein-coding genes, 20 tRNA genes and 2 rRNA gene fragments.The base composition shows a significant A+T preference (G2: 79.37%; G3: 79.41%).The 13 protein-coding genes use 5 types of start codons (ATG, ATA, ATT, ATC, GTG) and 2 types of stop codons (TAA, TAG).The phylogenetic analysis shows that G2 and G3 are supported by 100% bootstrap support to form an independent branch, which is located within the flea family but is significantly separated from known flea species.The G2 and G3 samples are on independent branches on the phylogenetic tree and have significant genetic distances from other species.The genomic length and base composition also differ from those of reported species.Based on molecular evidence, these two samples may represent a new undescribed species of the flea family, which awaits further morphological verification.This study reports the first mitochondrial genome of ectoparasites on the body surface of goats in the Qinghai-Xizang Plateau region, providing new data for the phylogeny and species diversity of the flea family.

Giardia duodenalis is a widespread zoonotic protozoan causing giardiasis in humans and livestock.A one-pot RPA-CRISPR/Cas12a assay was developed for rapid, visual detection of G. duodenalis for the detection of a specific genomic segment (GenBank: CP110919.1).For this crRNA and RPA primers were designed and optimized.The assay completed the detection within 30min at 37°C in a single tube, which demonstrated high specificity with no cross-reactivity against six nontarget pathogens.The limit of detection was 1 copy/μL for recombinant plasmids and 1 cyst per 10mL for cysts, surpassing nested PCR (10 3 cysts/10mL).In 30 clinical fecal samples, the assay showed 100% concordance with nested PCR.This one-pot system eliminates aerosol contamination risk and provides intuitive visual readouts, offering a field-applicable tool for giardiasis screening in cattle and goats.

In this study, reference materials (RMs) were developed to enable quantitative comparability for six major swine bacterial pathogens, including Actinobacillus pleuropneumoniae, Glaesserella parasuis, Streptococcus suis, Mycoplasma hyopneumoniae, Mycoplasma hyorhinis, and Salmonella enterica subsp.enterica serovar Typhimurium.These pathogens impose significant health and economic burdens on global swine production through severe respiratory diseases, systemic infections, polyserositis, arthritis, and zoonotic risks, resulting in increased mortality, reduced performance, and substantial treatment and vaccination costs.The optimized primer-probe assay targeted conserved genomic regions for each species to ensure broad detection capabilities across relevant strains using droplet digital PCR (ddPCR).RMs were prepared by combining bacterial genomic DNA with a swine tissue matrix mixture, followed by a comprehensive assessment of homogeneity and stability.The ddPCR-based approach demonstrated high reproducibility and sensitivity for homogeneity and stability assessment of RMs, providing absolute quantification without standard curves.Between-bottle homogeneity showed variations of 6.4-22.9%,representing the relative variation among aliquots observed during homogeneity assessment, while stability assessments confirmed reliability during storage at various temperatures and through multiple freeze-thaw cycles.Each RM was assigned a reference value with well-characterized measurement uncertainty.These RMs support quantitative comparability of analytical results under consistent analytical conditions and provide reliable reference values for evaluating diagnostic assays and standardizing molecular testing.However, further comparative evaluations across different analytical platforms, laboratories, and workflows are required to extend their applicability to diverse diagnostic environments.

Histological grade is an important prognostic factor in dogs with soft tissue sarcoma (STS) and is a key consideration in surgical planning.Complete surgical excision is the mainstay of treatment, and high-grade tumors generally require wider surgical margins and consideration of adjuvant therapy because of their higher risk of recurrence and metastasis.Accurate preoperative assessment of tumor grade is therefore important for treatment planning, but preoperative biopsy grading does not always match the final histological grade after surgery.Therefore, additional methods that can complement preoperative grade assessment are needed.This exploratory study investigated whether computed tomography (CT)-based radiomic features differed between low-and high-grade canine STS.Twenty-seven dogs with STS (20 low-grade [13 grade 1, 7 grade 2] and 7 high-grade [grade 3]) were included in this study.From preoperative contrast-enhanced CT images, 107 radiomic features were extracted according to the Image Biomarker Standardisation Initiative (IBSI) guidelines.Nine features were then selected through a three-stage process consisting of statistical filtering, exclusion of volume-dependent features, and hierarchical clustering.These features showed discriminatory performance for differentiating low-grade from high-grade tumors, with apparent AUC values of 0.843-0.957.GLDM-SmallDependenceLowGrayLevelEmphasis had the highest apparent AUC of 0.957 (95% CI: 0.864-1.000).Low-grade tumors tended to show higher values for low-gray-level texture features, whereas high-grade tumors showed higher values for features related to heterogeneous high-gray-level texture patterns.These findings suggest that CT-based radiomics may serve as a complementary tool for preoperative grade assessment in canine STS.Although these findings are encouraging, the reported AUC values reflect exploratory performance within this study cohort, and future prospective studies with larger cohorts and external validation are needed to confirm these findings.

The wildlife-livestock interface is an important setting for the emergence and transmission of infectious and parasitic diseases, driven by environmental change, animal movement, shared grazing resources and livestock management practices.Central Asian rangelands, particularly those in Kazakhstan, provide an important context for examining these dynamics because migratory saiga antelopes (Saiga tatarica) share habitats with domestic livestock, including sheep.This review examines the ecological, epidemiological, host-related and molecular factors influencing disease emergence at the wildlife-livestock interface, using the saigasheep system as a case study within a One Health framework.Particular attention is given to Pasteurella multocida, gastrointestinal nematodes and Coenurus cerebralis, alongside the potential influence of host genetic susceptibility, climatic variability and shared environmental exposure.Available evidence indicates that spatial overlap and the occurrence of related pathogens in wildlife and livestock provide opportunities for cross-species transmission but do not, in themselves, establish transmission or its direction.The review highlights the importance of integrating ecological surveillance, molecular characterization, genomic epidemiology and temporally matched sampling to distinguish shared exposure from genuine pathogen exchange and sustained transmission.It further discusses the role of livestock management, conservation-based veterinary interventions and coordinated institutional collaboration in mitigating disease risks.An integrated One Health approach combining movement ecology, molecular diagnostics, host susceptibility assessment and targeted preventive measures is essential to strengthen disease surveillance, protect wildlife populations and support sustainable livestock production in shared rangeland ecosystems.