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Nile tilapia Oreochromis niloticus is a popular freshwater fish that has been extensively and intensively cultured worldwide. However, cryopreservation of its sperm especially pre-freezing procedure, has not been properly developed. Therefore, the study aimed to determine the best pre-freezing procedure for cryopreservation of Nile tilapia Oreochromis niloticus. The completely randomized design with five treatments and four replications were employed in this study. The tested treatments were T1= 4 oC → 0 oC → -4 oC → -10 oC → -79 oC → -196 oC, T2= 4 oC→ 0 oC → -4 oC → -10 oC → -196 oC, T3= 4 oC → 0 oC → -4 oC → -196 oC, T4= 4 oC → 0 oC → -196 oC), and T5= 4 oC → -196 oC, with a 10 min equilibration at each respective temperature. Furthermore, sperm was cryopreserved for two weeks in liquid nitrogen (-179 oC). The results of the ANOVA test showed that pre-freezing had a significant effect on sperm motility, and viability (P<0.05), but had no considerable impact on sperm abnormality (P>0.05). Treatment T4 exhibited higher motility and viability, but these values were not significantly different from T3 and T5. Based on practical consideration, it was recommended to utilize the T5 pre-freezing procedures (4oC → -196oC) for cryopreservationof Nile tilapia sperm. Considering these results, Nile tilapia sperm can be directly cryopreserved into liquid nitrogen after equilibration at 4 oC for 10 min.

Traumatic ulcers are oral mucosa lesions caused by thermal, mechanical, and chemical factors. Long-term topical medication use can lead to fungal resistance. Patin fish skin contains glycine and glutamic acid, which can enhance macrophage activation and fibroblast proliferation, aiding wound healing. This study aimed to determine the effect of Patin skin gelatin on increasing macrophages and fibroblasts and reducing the diameter of traumatic ulcers. An experimental laboratory study with a post-test-only control group design was conducted. Ulcers were induced on the labial mucosa of the mandible in rats using thermal induction. Treatments included a placebo gel and Patin skin gelatin at concentrations of 10%, 20%, and 30% for 7 days. Rats were euthanized on days 3, 5, and 7, and ulcerated mucosal tissue was biopsied to count macrophages and fibroblasts using Olympus software. Data were analyzed with IBM SPSS Statistics 25. Results showed significant macrophage and fibroblast count differences between the gelatin treatment groups and the placebo. Patin skin gelatin at all concentrations increased the number of fibroblasts and decreased ulcer diameters significantly compared to the control group (p<0.05). Patin skin gelatin effectively enhanced fibroblast cell proliferation and reduced ulcer size over the 7-day treatment period.

Cryptosporidium spp. is a protozoan parasite of many vertebrates worldwide including avian hosts, causing gastroenteritis and diarrhea, although, studies have been conducted on Cryptosporidium spp. in some avians however, there is no information on Cryptosporidium spp. in pigeons from Anhui Province, China. To investigate the prevalence and assess the transmission burden of Cryptosporidium species in domestic pigeons, a total of 376 fecal samples were collected. The acid-fast staining and nested PCR amplification methods were utilized to detect the presence of Cryptosporidium oocysts. Microscopic examination and nested PCR reveal the prevalence rate of 5.05% (19/376) and 1.86% (7/376) respectively. Furthermore, molecular characterization was identified as Cryptosporidium meleagridis. As this study is the first report on Cryptosporidium spp in domestic pigeons in Anhui Province, we expect it to provide baseline information for further studies.

This study was conducted to determine the effects of Echinacea purpurea (EP) root on the growth performance and some haematological parameters of juvenile common carp (Cyprinus carpio). Echinacea meal was incorporated into the diets as follows: no Echinacea meal (E-0), 5 g Echinacea meal kg−1 diet (E-5), 10 g Echinacea meal kg−1 diet (E-10) and 20 g Echinacea meal kg−1 diet (E-20). Triplicate groups of fish averaging 9.6 ± 0.5 g were hand-fed twice daily for 60 days. At the end of the feeding trial, weight gain (WG), feed conversion ratio (FCR), and specific growth rate (SGR) ware significantly higher in fish fed the Echinacea-supplemented diets than in those fed the control diet. Red blood cell count (RBC) and white blood cell count (WBC) were significantly higher in the E-10 and E-20 groups than in the E-0 group. Fish fed the E-10 diet had the highest haematocrit (Hct), and haemoglobin (Hb) among all treatments. The nitroblue tetrazolium (NBT) assay showed a significant increase in the E-10 and E-20 groups compared to the E-0 and E-5 groups. After a 14-day challenge with Aeromonas hydrophila, Kaplan–Meier analysis revealed significant differences in survival between treatments. Survival of carp following the challenge was significantly higher in fish fed the E-10 and E-20 diets than in the E-0 group, whereas the E-5 group showed no significant difference compared to the E-0 group. In conclusion, our findings revealed that supplementing diets with EP root (10 g EP kg−1 and 20 g EP kg−1 diet) can enhance growth performance, haematological parameters, and resistance to A. hydrophila in juvenile common carp.

The objective of this research is to analyze key mineral parameters in Algerian Arbia goats during different reproductive phases to evaluate the variation in these parameters and their influence on some reproductive performance. The study was conducted at a Technical Livestock Institute (I.T.E.L.V) of Ksar Chellala in Tiaret, Algeria, from October 2022 to May 2023. During all stages of reproduction (estrus period, pregnancy, and post-partum period), 21 primiparous and multiparous Arbia goats were studied, aged between one and ten years and weighing 24,19 ± 6,08 kg at mating. Plasma calcium, phosphorus, magnesium, and iron, were estimated using the DIRUI CS-T180 Auto-chemistry Analyzer. Plasma sodium, potassium, and chloride levels were measured with ion-selective equipment (Easy Lyte ® Plus, MEDICA). In this study, mineral levels fluctuate in both groups: in primiparous and multiparous females, there is no appreciable difference (P>0,05) for electrolytes Na, K, and Cl throughout the various reproductive phases. However, P, Ca, and Fe concentrations were significantly lower (P<0,05) in the last month of pregnancy; also, Fe levels were significantly lower (P<0,05) in the post-partum phase. Nevertheless, the mineral plasma parameters during the late pregnancy stage and the age and weight of the goats at mating have a significant relationship. The multiparous kids (n=7) had a higher birth weight than the primiparous kids (n=8) (3,12 ± 0,54 and 2,84 ± 0,78, respectively). In Arbia goats, primiparous females were more fertile (100%), more productive (72,72%), and more prolific (114,28%) compared to multiparous females. Finally, it is found that parity influences the weight of the progeny at birth as well as the mineral concentrations at various stages of reproduction in Arbia goats. On the other hand, the reproductive characteristics of Arbia goats decline with parity.

This study is a continuation of innovative research on the impact of the simultaneous use of standard- and small-cell combs in bee colonies on the characteristics of worker bees and bee colonies. The nests of these colonies had two types of combs: small-cell (approximate cell width/size of 4.90 mm) and standard-cell (approximate cell width/size of 5.50 mm). The aim of the study was to compare the activities of proteases and their inhibitors present on the cuticle of workers reared in small-cell combs (SMC workers) and standard-cell combs (STC workers) in colonies kept simultaneously in standard- and small-cell combs. The width/size of comb cells in which the workers were reared had a significant effect on protein concentrations and activities of the proteolytic system, i.e. proteases and their inhibitors, on the cuticle surface. Regardless of the age of the workers (1 day, 7 days, 14 days, 21 days, and 28 days), the protein concentrations were statistically significantly higher (p ≤ 0.01) in the STC than SMC workers. The opposite was found in the case of the activities of proteases and their inhibitors: regardless of the age of the bees, the activities were significantly higher in the SMC workers. The differences between workers reared in small-cell combs and those reared in standard-cell combs may be responsible for their predispositions to perform different tasks in the colony. In our opinion, workers reared in small-cell combs are more predisposed to work as foragers outside the nest. However, this hypothesis requires confirmation in further research.

A change in testicular volume can indicate decreased male fertility and is useful for comparing spermatogenesis before and after experiments in animals, or before and after treatment of especially valuable domestic breeding animals with pharmaceuticals affecting spermatogenesis, such as antitumor drugs. Various formulae can be used to calculate the volume of living males’ testes. This study examined the suitability of four testicular volume calculation formulas for male Wistar rats of different ages (2-months, 1-year, and 2-years-old) and the changes in testicular volume during 90 days of observation. In 2-months-old rats, the solid ellipsoid triaxial body calculation formula and the prolate ellipsoid formula were most suitable for the calculation of testes’ volume, and the volume of both testes increased very significantly during the 90 days of observation. In 1-year-old rats, the solid ellipsoid triaxial body calculation formula was optimal, and the volume of both testes increased insignificantly. For 2-years-old rats, the solid ellipsoid triaxial body calculation and prolate ellipsoid formulas or their average was recommended, and the volume of both testes decreased slightly during 90 days of observation.

Lumpy skin disease (LSD) is a viral infectious disease that affects cattle and causes significant economic losses, particularly in low and medium income countries, where livestock is often the main source of income for small-scale farmers and rural communities. In the last few years, the disease has become widespread in several countries in Asia, including Pakistan. The present cross-sectional study aimed to determine the seroprevalence and molecular characteristics of LSD virus (LSDV) among cattle in the Bahawalpur district (Pakistan), while identifying potential associated risk factors. A total of 400 serum samples were collected from cattle and analysed using a commercial ELISA kit to determine seroprevalence. Additionally, 12 skin scrapping samples of cattle were collected from sick animals to detect and characterize the currently potentially circulating LSDV strains using PCR, targeting the P32 gene. The overall LSD seroprevalence among cattle was found to be 38.0%, with significant variation observed between different geographical areas of the Bahawalpur district, showing the highest prevalence of 46.2% in Yazman Tehsil. Age and feeding system were identified as significant risk factors for LSD occurrence in cattle. The genetic analysis revealed a high genomic similarity between the LSDV strain sequences reported in Asian and Middle Eastern countries. The P32 gene phylogenetic analysis further confirmed the close relationship between LSDV sequences from Pakistan and vaccine strains of sheep and goat pox viruses. The present study provides important baseline information for an understanding of the epidemiology and characterisation of LSDV enzootic strains in Pakistan, and highlights the need for effective disease control strategies, including vaccination campaigns, particularly in disease-endemic regions.

The aim of the study was to determine the effect of oral administration of selenitetriglycerides on the selenium level and selected hematological and biochemical parameters in Kamieniecka sheep. Sheep in the experimental group received a preparation in the form of selenitetriglycerides orally at a dose of 2 ml per animal for the next 7 days (1 mg Se/kg body weight). Blood was collected from each mother sheep: on day 0 before Se administration, and then on days 1 st , 7 th , 14 th , 21 st , and 30 th of the experiment. Serum selenium concentration in the control group was similar on all sampling days and ranged from 75.38 µg/l to 88.34 µg/l. In the experimental group, the selenium level was the lowest before supplementation and increased significantly during supplementation (p ≤ 0.01), starting from the 1st day of the experiment, to reach the peak value on the 7 th day (791.72 µg/l) and gradually decreased from another blood sample collection. The selenium concentration was significantly higher (p ≤ 0.01) compared to the baseline values even on the 30 th day of the experiment (214.78 µg/l) - 23 days after the end of selenitetriglyceride supplementation. The research showed that the dose of selenitetriglycerides was safe and effectively improved the Se level in the tested Kamieniecka sheep. Monitored hematological and biochemical parameters showed that supplementation does not negatively affect any vital functions of the body. Analyzing the above, it was found that oral administration of selenitetriglycerides is an effective and safe form of selenium supplementation in sheep mothers before reproduction.

The control of respiratory infections is integral to a healthy livestock farm. The current study was conducted to optimize a multiplex PCR assay for simultaneous detection of Pasteurella multocida (P. multocida), Staphylococcus aureus (S. aureus) and Mycobacterium bovis (M. bovis) in nasal samples of cattle having suspicion of respiratory tract infections. The nasal samples were collected from 10 dairy farms affected with respiratory disease outbreaks in the recent past. These outbreaks were associated with respiratory tract infections caused by bacterial pathogens P. multocida, S. aureus and M. bovis. A multiplex PCR assay was therefore optimized to enable the simultaneous detection of these bacterial pathogens directly from nasal samples. The multiplex PCR assay was optimized using primers already validated for efficient amplification of specific DNA segments in reference strains of targeted bacterial pathogens. The standardized assay was specific and sensitive enough to detect ≥ 100 genome equivalents of target DNA segments in each of P. multocida, S. aureus and M. bovis. The assay was successfully applied for the detection of the three bacterial pathogens in 50 nasal samples. PCR amplicons were subjected to Sanger dideoxy sequencing followed by phylogenetic analysis to determine if species identification was specific. In short, the optimized multiplex PCR assay may prove as a reliable, economical, and simple tool for the management of bovine respiratory tract infections caused by some key bacterial pathogens.

Porcine reproductive and respiratory syndrome (PRRS) is caused by the PRRS virus (PRRSV). It leads to reproductive disorders and respiratory failure in sows and piglets. As an endoribonuclease, the PRRSV non-structural protein 11 (NSP11) is crucial in replication and assists immune system evasion. NSP11, which is relatively more conserved than NSP2, could potentially cause a new round of PRRSV epidemics, given a major mutation occurs. Here, we aimed to analyze the PRRSV-2 NSP11 genetic diversity in China between 1996–2022. The NSP11 nucleotide sequence was analyzed in 60 PRRSV-2 strains, revealing a similarity of 83.6%–100%. Similarly, amino acid sequences exhibited homology ranges of 91.0%–100.0%. Amino acid sequence alignment analysis re-vealed multiple substitutions in NSP11. NSP11 phylogenetic analysis of 489 PRRSV-2 strains revealed that Lineages 8 and 1 were the predominant strains of PRRSV circulating in China. These two lineages exhibit relatively close genetic relationships. Although unsupported by SimPlot analysis, recombination analysis suggested a potential recombination event in the 489 PRRSV-2 NSP11 sequences. Recombination analysis and amino acid sequence alignment confirmed the PRRSV NSP11 conservation. Our findings provide genetic diversity of PRRSV-2 NSP11 in China and contribute to effective strategy development to prevent and control PRRSV.

Postpartum inactive ovaries (IO) in dairy cows reduce the economic returns of the dairy industry. It is related to energy metabolism disorder, hormone levels and cytokines.The aim of this study was to evaluate the correlation between insulin resistance (IR), adiponectin (ADPN), and leptin (LEP) at 14 days postpartum to assess the predictive potential for IO risk in dairy cows. Cows at 14 days postpartum were randomly selected and allocated into an insulin resistance group (IR, with IR index > 2.5, n = 30) and a non-insulin resistance (non-IR, with IR index < 2.5, n = 30). Serum Samples were collected at 14 and 55 days postpartum. Six cows of estrus and six cows of IO were randomly selected for slaughter at 55 days postpartum. Then, adipose and ovary samples were allocated for further experiments. A significant association between IR and IO, with 53.33% prevalence in the IR group compared to 16.67% in the non-IR group. Cows with IR had higher levels of β-hydroxybutyrate, non-esterified fatty acid, and lower levels of glucose, total cholesterol, triglyceride, ADPN, and LEP. Reproductive performance was adversely affected, with IR cows showing longer durations for first estrus and reduced milk yield. ADPN and LEP levels were significantly lower in IR cows, suggesting their role in modulating insulin sensitivity and reproductive functions. The combined analysis of ADPN, LEP, and IR index showed high sensitivity (91.3%) and specificity (87.2%) in predicting IO, highlighting their potential as reliable biomarkers. These observations indicate that IR and serum LEP and ADPN at 14 days postpartum can predict IO in dairy cows.

This study aimed to evaluate the impact of body condition score (BCS) on the milk yield, reproductive performance, and health status of lactating dairy cows. Data were collected from 1,960 cows across four dairy farms at 21 days prepartum, on the day of calving, and at 21, 50, 150, 200, and 250 days postpartum. The dataset included BCS, lactation performance, reproductive performance, disease incidence, and economic benefits for each cow. The cows were divided into seven groups based on BCS: ≤ 2.5, 2.75, 3.0, 3.25, 3.5, 3.75, and ≥ 4, with 40 cows per group at each time point (ten cows were selected from each of the four farms based on BCS differences at each time point for the experiment). The results demonstrated significant differences in BCS, milk yield, reproductive performance, and disease incidence among cows from different dairy farms. From 21 days prepartum to the day of calving, cows with BCS of 3.25 and 3.5 exhibited superior milk yield, reproductive performance, and economic benefits, as well as lower disease incidence. From 21 to 250 days postpartum, cows with BCS values of 3.0 and 3.25 continued to show improvements in milk yield, reproductive performance, and economic benefits, along with a reduction in disease incidence. These findings suggest that the ideal BCS for cows at 21 days prepartum and on the day of calving is 3.5, while BCS of 3.25 is optimal for cows at 21 days postpartum, and BCS of 3.0 is recommended for cows from 50 to 250 days postpartum.

Honey is natural nectar that honey bees gather from a variety of flowers. Honey is made up of many nutrients that are essential for the growth and development of various tissues and organs in all kinds of living organisms. The goal of the current study was to determine whether adding honey to diets of naked neck cocks would have a good impact on their semen quality, haematological profile, and immune system. A total of 90 naked neck cocks that were 75 weeks old were gathered for this purpose in March and April 2021 from the University of Agriculture Birds Stock. The birds were divided into 3 equal treatment groups: control, A and B, which each received a dose of 5 and 10 g of honey in 1 litre of water (w/v), respectively. The control group received no treatment. Five weeks of treatment were given, with one week serving as an adaptation period. One-way analysis of variance was applied to the recorded data for statistical analysis. The addition of honey to drinking water at a dose of 10 g (w/v) increased (p<0.05) sperm motility, followed by a dose of 5 g (w/v) and the control group, but the mean values of non-motile sperms were lowest (p<0.05) at a dose of 10 g (w/v), followed by a dose of 5 g (w/v), and the control group. However, there were no significant differences in semen volume, pH, or concentration across the treatment groups. The haematological parameters, such as haemoglobin, total leukocyte count, red blood cells, and packed cell volume, were not significantly affected by the addition of honey to the drinking water, but the ND titer of naked neck cocks treated with 10 g (w/v) honey was significantly (p<0.05) higher than that of the cocks in the 5 g (w/v) and control group. In conclusion, the addition of honey to drinking water at a dose of 10 g (w/v) may have positive effects on sperm motility and antibody titer against Newcastle disease in aged naked neck cocks. Furthermore, these results also suggest that the addition of honey to drinking water at a dose of 10 g (w/v) may rejuvenate naked neck cocks even at old age (75 week age).

Bovine mastitis is primarily treated with antimicrobial agents. Anti-inflammatory agents are also used to alleviate clinical symptoms or reduce antimicrobial use. Glycyrrhizin is an anti-inflammatory agent used in the treatment of bovine mastitis, but its effects are not fully understood. We therefore examined the anti-inflammatory effects of glycyrrhizin both in vivo and in vitro. We first tested whether glycyrrhizin exerts anti-inflammatory effects using MAC-T cells, an immortalized bovine mammary epithelial cell line. Glycyrrhizin decreased the expression of interleukin (IL)-1β mRNA in a concentration-dependent manner in MAC-T cells stimulated with lipoteichoic acid (LTA). We then investigated the effects of glycyrrhizin in bovine mammary epithelial cells (bMECs), which seem to retain more of the characteristics of actual mammary epithelial cells. Stimulation with LTA or lipopolysaccharide significantly increased cytokine mRNA expression in bMECs. Glycyrrhizin exhibited a slight inhibitory effect, but no significant difference was observed. The effect of glycyrrhizin on LTA-induced mastitis was examined in lactating cows. Quarters were divided into test and control areas (test quarter: n=8, control quarter: n=7). All quarters were stimulated with LTA at the start of the trial (0 h). In the test quarter group, glycyrrhizin was administered via intramammary infusion. The somatic cell count and relative gene expression of IL-1β and tumor necrosis factor–α were significantly lower in test quarters than control quarters. Both the in vitro and in vivo studies showed that glycyrrhizin reduces the expression of proinflammatory cytokine genes in response to LTA-induced inflammation and partially revealed the mechanism of the anti-inflammatory effect of glycyrrhizin on mastitis. Further investigations involving field cases of mastitis with bacterial infections are needed to demonstrate the anti-inflammatory effect of glycyrrhizin on bovine mastitis.

An animal body's immune response to viruses might vary depending various factors. Herein, the relationship between 2'-5'-oligoadenylate synthetase like (OASL), interferon alpha (IFN-α), and duck hepatitis A virus type 3 (DHAV-3) virus genome copy number in duck embryo tissues was studied to investigate duck's natural antiviral immunity mechanism.15-day-old SPF duck embryos were infected with DHAV-3 and their organs and tissues were collected at various times after inoculation. RT real-time PCR was used to determine OASL and IFNA mRNA expression levels and the DHAV-3 copy number. Compared with that at 0 hours, liver OASL transcription increased significantly at 24 hours, and extremely significantly in the liver, heart, stomach, intestine, and muscle at 48 hours. The heart had the highest expression level, followed by the liver, with lower expression in the other tissues. There was no significant difference in IFNA expression between the heart and liver at 12 hours, but it started to rise at 24 hours, reaching its maximum at 48 hours. Although IFNA expression increased in the stomach, intestines, and muscles, its relative expression levels were lower than those in the heart and liver. In the tissues, the virus genome copy number peaked at 24 hours, and then decreased. The liver had the highest virus genome copy number, followed by the heart. The results implied that the differences in OASL transcription in the tested tissues were similar to those of IFNA. Thus, the immune response (OASL and IFN-α levels) of duck embryos to DHAV-3 shows tissue differences, especially in the liver and heart.

The purpose of this study was to compare the effect of the age of the breeder flock of commercial BUT - 6 turkeys on the transfer of maternal antibodies to chicks. The blood samples for serological analysis were collected from randomly selected 63 female breeder from flock of BUT Big 6 turkeys and 63 one-day-old hybrid turkey poults hatched from eggs from this flock at 36, 45 and 54 week of age. During blood analysis (serum) in the laboratory, the level of antibodies of the breeder flock against Avian metapneumoviruses (APV), Newcastle disease virus (NDV) and Hemorrhagic enteritis virus (HEV) was determined (ELISA). Maternal antibody (MatAb) titer in chicks (serum) against the same viruses were also determined. The percentage (%) transfer of MatAb to offspring was then evaluated. The effect of the age of the turkeys on the antibody titer to the tested pathogens expressed in geometric mean titers (GMT) was shown. During the laying period, the antibody titer of the tested turkeys against NDV decreased with the age of the flock. The highest antibody titer was demonstrated in 36wk. (GMT=14242), whereas the lowest in 54wk. (GMT=5564). In contrast, serum antibody titer of the tested layers against APV and HEV increased with the age of the birds. The lowest antibody titer (GMTAPV=24818; GMTHEV=12070) was observed at the beginning of the laying period, and the highest at the end of the laying period (GMTAPV =38978; GMTHEV =13980). The highest vertical transfer to offspring was shown for antibodies to – HEV (82.7%), while the lowest was shown when analyzing sera to -NDV (37.6%). The present analysis showed significant differences in the evaluated antibody titres in serum of turkey breeders during the laying period, as well as in the level of MatAb in chicks. The presented results also showed that the transfer of MatAb to chicks is influenced by the age of the parent flock and the type of pathogen against which the layers were vaccinated.

In this study we investigated the antioxidant status of fractionated seminal plasma (SP1, >40 kDa; SP2, <40 kDa), obtained by gel filtration chromatography (Wasilewska-Sakowska et al. 2019a), and mitochondrial functions in SP-treated spermatozoa during pre-freeze equilibration and post-thaw storage periods. The Total Antioxidant Status (TAS), represented by enzymatic and non-enzymatic antioxidants was markedly affected by the treatments, being significantly higher in either SP1 or SP2 than in the whole seminal plasma (wSP). It was observed that ATP content was markedly higher in SP1- or SP2-treated samples following pre-freezing equilibration. Although there were no marked differences in the ATP content of frozen-thawed (FT) spermatozoa among treatments, regardless of the cryopreservation stage, mitochondrial membrane potential (MMP) of FT spermatozoa differed significantly among treatments. Furthermore, significant differences in the sperm MMP at post-thaw were not reflected in the ATP levels, suggesting that variations in the surrounding environments of FT spermatozoa might have varying effects on the sperm mitochondrial functions following cryopreservation. It seems that high TAS levels in the fractionated SP could reaffirm the protective role of antioxidants in sperm cryo-damage.

Enzootic bovine leukosis (EBL) is the most prevalent neoplastic disease in dairy and beef cattle, caused by the Bovine Leukemia Virus (BLV). BLV can infect a wide range of animal species, however, cattle and buffalo are considered natural hosts for BLV. This study aimed to determine the molecular prevalence of BLV in different cattle farms of Khyber Pakhtunkhwa and the prevailing genotypes of BLV by sequencing the partial gp51 gene. A total of 1250 blood samples were collected from cattle of each sex, various age groups, breeds, etc from different cattle farms. Out of a total 1250 samples, BLV was detected in 136 samples (10.88%) through nested PCR. The risk factors analysis revealed that BLV was significantly higher in exotic breeds and cattle having a high age group. The transmission of BLV in the herd may be horizontal i.e., through contaminated equipment, close contact with infected animals, increased stocking density, or blood-sucking insects, or vertical through artificial insemination or natural breeding. A total of four PCR-positive samples were sequenced for confirmation and genotyping of BLV isolates. The phylogenetic analysis revealed that the isolates belong to genotype I and were in proximity with GI BLV isolates from Japan. Moreover, the current study isolates were tightly clustered together which may point to their origin from the earliest virus that was previously introduced to the hosts in the study area. The global human and animal movement, and purchasing of cattle from BLV endemic countries i.e., USA, Japan, Australia, etc have significantly contributed to the transmission of BLV.