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 Trypanosoma evansi in Southern Punjab, Pakistan, focusing on key ruminant species, including camels, cattle, buffaloes, goats, and sheep.  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 T. evansi.  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. evansi 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.  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. 

Bluetongue is a vector-borne, emerging disease that poses a severe threat to most domesticated animals. A cross-sectional study was conducted to estimate the prevalence of bluetongue virus (BTV) disease in apparently healthy sheep and goats in Kalat, Balochistan.  A total of 270 serum samples (sheep: 150 and goat: 120) were collected and screened for the detection of anti-BTV antibodies using a competitive enzyme-linked immunosorbent assay. The data regarding different contributory risk factors were also collected using a predesigned questionnaire.  It revealed that overall, 27.4% (74/270, 95% confidence interval, χ2 = 1.71, p = 0.12) prevalence in both sheep and goat populations. The highest prevalence of 47% (32/68) was recorded in Surab city with the lowest prevalence of 15.49% (11/71) in the Manguchar area. In contrast, in Kalat 28.1% (9/32), Daan area 24% (12/50), and Marap area 22.44% (11/49), seropositivity was recorded. Upon sex bases, antibodies were almost equally found in both male 28.57% (8/28) and female 27.27% (66/242) animal populations. Moreover, all four breeds (Balochi, Khurasani, Lehri, and Rakhshani) were equally and potentially seropositive. The Khurasani breed was the most susceptible to 34.69% (17/49), followed by the Balochi breed, 45/145 (31%) seropositivity. The prevalence of BTV was 16.66% (1/6) in Rakhshani breed and 15.71% (11/70) in Lehri breed., Ticks were found in almost 21% of animals, while 93% of animals were reared on open grazing in rangelands.  This study clearly indicates widespread BTV infection in small ruminants in the study area that may pose serious threats to livestock farming. Further extensive studies are recommended to study the prevalence of disease in different agroecological zones of the province. This also warns the high-ups to manage concrete efforts to eradicate and control the disease in the area. 

Alzheimer’s disease (AD) poses a significant health-care challenge, often linked to cognitive decline caused by oxidative stress. This study investigated the potential neuroprotective effects of the Paederia foetida leaf extract (PFE) in rats that exhibited scopolamine-induced dementia mimicking AD.  Forty-two male rats were treated with either donepezil (0.5 mg/kg) or PFE at doses of 250, 500, and 1000 mg/kg for 14 days before and 14 days after the beginning of Alzheimer’s-like symptoms after 14 consecutive days of scopolamine administration. Behavioral tests, including the open-field test for locomotor activity and the Morris water maze task for learning and memory assessment, were conducted. Neuronal cell counts and biochemical assays were performed to further analyze outcomes.  All groups exhibited normal locomotor activity. The scopolamine group displayed longer escape latency times, reduced time in the target quadrant, decreased number of surviving neurons, and increased malondialdehyde and decreased glutathione levels compared with the control group. However, pre-treatment with 1000 mg/kg PFE notably mitigated the neurotoxic effects of scopolamine.  The neuroprotective properties of PFE are highlighted, suggesting its potential as a promising treatment strategy for AD. 

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.  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.  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.  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. 

Ducks worldwide are infected with duck circovirus (DuCV), which causes feather abnormality, emaciation, and poor growth performance. DuCV is similar to other circoviruses that induce immunosuppression due to the occurrence of the bursae of Fabricius (BF) and spleen atrophies. In Thailand, retarded ducks with feather losses were submitted for disease investigation. The ducks presented low body weight gain, had small BF and spleens, and were consistent with duck-infected DuCV. Our study investigated the possibility of DuCV infection in duck flocks in Thailand. We also analyzed the genetic characteristics of the virus.  BF and spleen samples were collected from affected meat and layer ducks from six farms thought to have been infected with DuCV. These tissues were then subjected to histopathological examination and molecular identification using conventional polymerase chain reaction and nucleotide sequencing. To identify DuCV, phylogenetic trees were generated using MEGA version X software. Samples of tissues or swabs were collected to determine whether coinfections with bacteria and viruses existed.  Phylogenetic analysis using the entire genome (1995–1996 bp) and cap gene (762 bp) revealed that the DuCV isolates circulating in Thailand belonged to DuCV genotype I, which was further subdivided into two sub-genotypes: sub-genotype I b and an unclassified sub-genotype based on reference sub-genotypes. Thai isolates have variations in 10 amino acid residues in the capsid protein. Ducks infected with Thai DuCV were also coinfected with Riemerella anatipestifer, Escherichia coli, Pasteurella multocida, duck viral enteritis, and duck Tembusu virus, which is consistent with previous DuCV infection studies.  Six DuCVs from ducks who were previously found to have feather loss, were underweight, had growth retardation, and had poor body condition were identified in this study as belonging to genotype Ⅰ and constituting at least two sub-genotypes. Due to the immunosuppressive effects of DuCV, coinfection of bacterial and viral pathogens was typically observed in Thai DuCV-infected ducks. 

The sun conure (Aratinga solstitialis), a bird belonging to the Psittaciformes family, is a popular pet because of its bright color and beautiful appearance. The sun conure is a monomorphic bird with similar appearances between males and females, making sex identification difficult by observing the external morphology. Therefore, molecular techniques are utilized. Loop-mediated isothermal amplification (LAMP) is a molecular technique that is often applied for sex identification in birds and is a quick and simple method that can be used in the field. This study used the LAMP technique to improve sex identification in sun conures by observing the color change of hydroxy naphthol blue.  Two primer sets, SunSpin-W and SunSpin-Z, were designed for sex identification in sun conures using the LAMP technique specific to the spindlin gene. The developed LAMP reaction was tested for optimal conditions, sensitivity, and specificity compared with the polymerase chain reaction (PCR) technique.  The SunSpin-W primer set amplified only female birds, whereas the SunSpin-Z primer set amplified DNA from both male and female birds. The primer sets were optimized at 62°C for 45 min. A positive result was visible to the naked eye from the color change of the reaction. In the LAMP assay, the lowest detectable concentration was 10 pg/μL and in the PCR assay, it was 1 ng/μL, while a 100% accuracy rate in sex identification was observed when comparing the LAMP assay results with the PCR assay.  This study successfully developed a LAMP technique for sex identification of sun conure, which took 45 min to complete and can be expanded for use in the field. 

The dearth of new antibiotics necessitates alternative approaches for managing infections caused by resistant superbugs. This study aimed to evaluate the lytic potential of the purified bacteriophage PKp-V1 against extended-spectrum β-lactamase (ESBL) harboring hypervirulent Klebsiella pneumoniae (hvKp)-K1 recovered from veterinary specimens.  A total of 50 samples were collected from various veterinary specimens to isolate K. pneumoniae, followed by antimicrobial susceptibility testing and molecular detection of various virulence and ESBL genes. Multilocus sequence typing of the isolates was performed to identify prevalent sequence types. The bacteriophages were isolated using the double-agar overlay method and characterized using transmission electron microscopy, spot tests, plaque assays, stability tests, and one-step growth curve assays.  Among 17 (34%) confirmed K. pneumoniae isolates, 6 (35%) were hvKp, whereas 13 (76%) isolates belonging to the K1 type were positive for the wzy (K1) virulence gene. All (100%) hvKp isolates exhibited the allelic profile of ST258. Overall, PKp-V1 exhibited an 88 % (15/17; (p ≤ 0.05) host range, among which all (100 %; p ≤ 0.01) hvKp isolates were susceptible to PKp-V1. PKp-V1 exhibited a lytic phage titer of 2.4 × 108 plaque forming unit (PFU)/mL at temperatures ranging from 25°C to 37°C. The lytic phage titers of PKp-V1 at pH = 8 and 0.5% chloroform were 2.1 × 108 PFU/mL and 7.2 × 109 PFU/mL, respectively.  Although the incidence of ESBL-infected K. pneumoniae in veterinary settings is worrisome, PKp-V1 phages showed considerable lytic action against the host bacterium, indicating the potential of PKp-V1 as a possible alternative therapeutic option against MDR K. pneumoniae. 

Extended-spectrum β-lactamases (ESBLs) are β-lactamase enzymes produced by Gram-negative bacterial pathogens that harbor the ESBL genes. In addition, most ESBL genes are plasmid-mediated and usually encode a broader spectrum of antimicrobial resistance, especially to penicillins, first-generation, second-generation, and third-generation cephalosporins, as well as monobactam, such as aztreonam. Escherichia coli has become an opportunistic pathogen, especially in poultry, and has been implicated in zoonotic diseases that can be transmitted to humans, resulting in public health problems. Poultry can act as carriers of ESBL-producing E. coli (ESBL-EC) bacteria to humans through poultry meat that is contaminated by waste products, feces, and excretions. The ESBL gene CTX-M type was identified as the main cause of infection in humans and was detected in poultry as a cause of infection accompanied by clinical symptoms. Several studies have also shown a link between E. coli and ESBL gene transfer from birds to humans. Controlling the spread of ESBL-EC involves maintaining the cleanliness of poultry products, especially meat, and eliminating contaminant sources from poultry. Likewise, maintaining the environmental cleanliness of poultry slaughterhouses and poultry farms must be taken as a precautionary measure to curtail the increasing spread of ESBL-EC into the environment. This review aimed to explain the spread of ESBL-producing E. coli in poultry. 

Postpartum uterine disease, such as endometritis, is widespread in highly productive dairy cows, leading to fertility problems and economic losses. Despite existing diagnostic methods, early and effective detection of uterine infections remains problematic due to the subclinical nature of some conditions. This study aimed to develop and evaluate a rapid diagnostic test for endometritis in cows on different days postpartum (PP).  The study was carried out on dairy Holstein–Friesian cows (n = 121) kept on farms in northern Kazakhstan. The study included both clinically normal cows and those diagnosed with endometritis, ensuring a comprehensive evaluation of the diagnostic methods across different stages of uterine health. The following laboratory tests were performed to diagnose and evaluate the presence and severity of endometritis in the cows: Nagorny-Kalinovsky test, Whiteside test, Katerinov test, Gavrish silver nitrate test, peroxide test, and clinical tests (rectal and vaginal examination). These tests were specifically chosen to identify inflammatory processes in the uterus, particularly focusing on detecting endometritis.  From day 21 to 30 PP, rectal and vaginal examinations were 32% and 28% more effective than the Nagorny-Kalinovsky test and the Whiteside test. From day 61 postpartum, the Whiteside test was 37.5% more effective than the Nagorny-Kalinovsky test. Comparatively, among laboratory diagnostic methods from days 10 to 110 PP, the peroxide test showed the greatest effectiveness in identifying 80.9% of sick animals. In sick animals from day 10 to 20 PP, during the interaction of the cervicovaginal mucus with 10%, 20%, and 30% hydrogen peroxide (H₂O₂), an 8.1 ± 1.9–8.8 ± 1.6 cm foam column was formed within 4–5 min.  The experiment showed that a 10% H₂O₂ solution yielded better results. Using H₂O₂ as a diagnostic agent for endometritis in cows has several advantages, including ease of use, it does not require special laboratory conditions and provides a visual reading of the reaction within 4–5 min. A limitation of this study is the focus on H₂O₂ without exploring other potential reagents that may enhance diagnostic accuracy. Future research could explore the long-term stability of cervicovaginal mucus samples and investigate the integration of additional substances that may expedite the detection of subclinical endometritis and improve the clarity of diagnostic results. 

Fracture healing can cause serious complications, both preoperatively and postoperatively, including malunion or non-union. Biomaterials can enhance the fracture healing process. This study aimed to compare platelet-rich plasma (PRP)-chitosan and platelet-rich fibrin (PRF)-chitosan on the basis of biochemical parameters for fracture treatment in rabbits.  This study involved 12 clinically healthy rabbits. After preparing PRP and PRF, a 3-mm bone defect was created in the tibia of each rabbit. The animals were divided randomly into two groups (A and B). Group A received PRP-Chitosan, and Group B received PRF-Chitosan. Bone healing was assessed using biochemical parameters (calcium [Ca], phosphorus [P], serum alkaline phosphatase [ALP], and osteocalcin [Ocn]) at 2-, 4-, 6-, and 8-week postoperatively. The data were compared using repeated-measures analysis of variance (p < 0.05) with Statistical Package for the Social Sciences statistical software.  Group-wise comparison showed no significant differences (p > 0.05) between the groups, except for ALP levels, which were significantly higher in Group B than in Group A (p < 0.05). In the week-wise comparison, there was a significant difference between both groups, as Ca and ALP levels showed significant differences at all weeks postoperatively, whereas Ocn showed a significant difference at 2- and 4-week postoperatively (p < 0.05). However, there was no difference in P levels between the groups at any post-operative week (p > 0.05).  Both combinations enhanced bone regeneration. However, PRF-Chitosan is a better combination for bone repair than PRP-Chitosan. There were some limitations of this study, such as a small sample size, only male rabbits were used, and a lack of mechanical testing; these limitations should be addressed in future studies. The insights gained from the present study may open a new approach to the use of a combination of biomaterials for bone healing, which should be further investigated clinically and in other animal models as a future scope. 

Environmental stress poses serious threats to animal welfare and production, particularly in poultry, which are susceptible to such stress. It can increase susceptibility to diseases and infections, reduce growth rates and reproductive performance, and increase behavioral issues. Environmental stress caused by conventional housing conditions can negatively affect well-being and productivity. High temperature, overcrowding, poor ventilation, insufficient lighting, and wire cages are some of the most prominent stressors in conventional housing systems. To address environmental stress in chicken farms, some strategies and tools, such as using anti-stress feed additives and enriching cages, can help improve bird behavioral activities and welfare. Breeders can improve overall bird performance by implementing these strategies and creating a more enriched and comfortable environment. Thus, this review discusses the importance of using different feed additives and environmental enrichment materials to reduce stress in chicken farms (broiler and layer) and improve bird productivity and well-being. 

Cholangiocarcinomas are malignant neoplasms that originate from any part of the bile duct epithelium. It is one of the most common liver tumors in dogs. This study described the clinical, cytological, hematological, biochemical, and pathomorphological findings of five cholangiocarcinoma cases in exotic breed dogs aged 2–5 years to aid in clinical diagnosis.  This study used dogs presented at different times from 2012 to 2021 at the Veterinary Teaching Hospital, Federal University of Agriculture, Abeokuta. History, clinical signs, and vital parameters were recorded. Blood samples were collected for hematology and serum chemistry. Abdominocentesis was performed for cytological diagnosis. All dogs died during treatment, and postmortem examinations were performed. At postmortem, fine needle aspirates were collected from the liver and mesenteric lymph nodes and liver and kidney samples were fixed in 10% neutral-buffered formalin.  The dogs showed signs of severe malnutrition, jaundice, and bloating. The hematological analysis indicated anemia, neutrophilia without band neutrophils, and lymphopenia, indicative of a stress hemogram. The serum biochemistry test revealed lower levels of total proteins, albumin, and globulin and higher levels of serum enzymes. Abdominal fluid and mesenteric lymph node cytology revealed clusters of epithelial neoplastic cells. A postmortem examination revealed the liver’s nodular enlargement with the presence of button-like ulcers. Neoplastic epithelial cells are solid masses with hyperchromatic nuclei surrounded by fibrous connective tissues.  Cholangiocarcinoma, diagnosed over a period of time in five exotic breeds of dog, consistently presents with the same clinical and postmortem findings, aiding in clinical diagnosis. However, the diagnosis of the disease is not possible in the early stage because of the absence of specific clinical signs. In dogs and possibly other animal species presenting with emaciation, lethargy, icterus, and distended abdomen, cholangiocarcinoma should be suspected, and cytological examination of the abdominal fluid and lymph node aspirates should be performed despite the absence of advanced equipment. 

Cryptosporidiosis is a major waterborne disease affecting ruminants and humans worldwide. It causes diarrhea and neonatal mortality in buffalo calves, and watery diarrhea and mortality in children and immunodeficient patients. This study aimed to investigate the efficacy of Myristica fragrans methanolic extract in treatment of C. parvum infection in comparison with nitazoxanide (NZX) (a Food and Drug Administration-approved drug control) in immunosuppressed and immunocompetent mice.  One hundred laboratory-bred male Swiss albino mice were equally divided into immunocompetent and immunosuppressed groups. Each group was further divided into five subgroups: (1) non-infected and non-treated control, (2) infected and non-treated control (infected with Cryptosporidium parvum oocysts 3 × 10³), (3) NZX-treated (100 mg/kg, 200 μL/mouse), (4) M. fragrans Houtt. methanol extract-treated (500 mg/kg), and (5) combination-treated (NZX + M. fragrans extract). Number of oocysts/g of feces, serum immunoglobulin (Ig) G level, and interferon (IFN)-γ, and interleukin (IL)-4 levels were used to evaluate the therapeutic effect.  C. parvum oocyst shedding in stool samples was significantly decreased in all treatment groups, with 79.7%, 81.2 %, and 85.5 % reduction in immunocompetent mice treated with NZX, M. fragrans, and their combination, respectively. In immunosuppressed mice, oocyst shedding was reduced by 77.7%, 80.5 %, and 83.7 % upon NZX, M. fragrans, and their combination treatments, respectively. The serum IgG level was lowest in mice treated with a mixture of M. fragrans and NZX, followed by those treated with NZX, and was highest in mice treated with M. fragrans alone. Regarding cytokine levels, all groups treated with M. fragrans had low levels of IFN-γ and IL4 on day 21 post-infection.  Collectively, the treatment of cryptosporidiosis with M. fragrans extract was successful in mice, as demonstrated by the measured parameters. M. fragrans reduced C. parvum oocyst shedding and serum IgG, IFN-γ, and IL-4 levels in immunocompetent and immunosuppressed mice. 

Male fertility is essential to bovine reproduction, particularly when bulls are used for artificial insemination or single-sire breeding. Bull breeding and soundness examinations (BBSE) are routinely undertaken to identify potentially unfit bulls for breeding. Multiple criteria, including physical examination and determination of testicular and semen parameters, characterize BBSE. Knowledge interstices within this realm, especially in tropical African settings, necessitate pragmatic approaches to address the same. This study aimed to investigate the potential effects of bull parameters on testicular attributes in a tropical setting.  The present study recruited healthy bulls (n = 96) aged >16 months with active semen collection used for artificial insemination at the Kenya Animal Genetic Resources Center. The breed breakdown was as follows: Ayrshire (n = 40), Boran (n = 3), Friesian (n = 36), Guernsey (n = 5), Jersey (n = 7), and Sahiwal (n = 5). Age, breed, body weight, scrotal circumference (SC), and testicular echotexture were collected, and the findings were analyzed using R statistical software.  SC increased with age and body weight (p < 0.0001). SC varied from one breed to the others (p < 0.0001). Furthermore, as determined using trans-scrotal ultrasonography, hyperechoic testicular lesions were present in 30.21% of the bulls imaged, and the incidence was significantly related to age (p < 0.001).  SC is significantly affected by age and body weight. The mean SC was higher in the Bos indicus, but this finding is only indicative because the Sahiwal and Boran sample sizes were small. Notably, the prevalence of hyperechoic testicular foci following trans-scrotal ultrasonography was common in older bulls. However, there is a need to further elucidate this phenomenon’s pathophysiology with age as the etiology and possible sequelae of semen quality. 

Rice straw, a widely available agricultural byproduct globally, has significant potential as a basal diet for livestock. The major challenge lies in obtaining high-protein foliage that can be easily extracted using natural water rather than chemical solvents. This study aimed to assess the ability of distilled water to extract protein concentrate from Indigofera leaves (Indigofera zollingeriana Miq.) and to evaluate its effectiveness in enhancing rumen feed fermentation and digestibility in low-quality rice straw basal diets.  The study was conducted in two experimental series. Experiment 1 was designed to explore the ability of distilled water to extract protein concentrate from fresh and dry Indigofera leaves by comparing it with the 0.1 N NaOH standard solvent. Experiment 2 focused on the in vitro digestibility of protein concentrates extracted from fresh Indigofera leaves based on optimal findings from experiment 1. Five treatments consisting of 0.5% and 1.0% protein concentrate and two extractants (distilled water and 0.1 N NaOH) were used to extract protein from Indigofera leaves. These extracts were then added to rice straw-based diets. Rice straw without supplements was used as a control. The treatments were arranged using a randomized complete design with five replicates.  The results of experiment 1 showed that distilled water was superior to 0.1 N NaOH for extracting protein concentrate from fresh Indigofera leaves, as revealed by higher dry matter, protein yield, total amino acids (AA), and total essential AA (EAA) production. For in vitro experiment 2, supplementation with distilled water-extracted protein concentrates successfully increased rumen fermentation and digestibility in rice straw basal diets, as indicated by higher gas production, total volatile fatty acid, and microbial protein levels compared with 0.1 N NaOH.  Findings from this study confirm that Indigofera leaf protein concentrate offers a new alternative for enhancing rumen feed fermentation and the digestibility of low-quality rice straw diets. This study implies that it is an easy, cost-effective, and environmentally friendly approach, particularly beneficial for smallholders, to extract protein concentrate from fresh Indigofera leaves using distilled water and use it to enhance the quality of rice straw for ruminant feed. The limitation of this study is that the Indigofera supplement was established using in vitro digestibility under controlled laboratory conditions, which does not reflect real rumen conditions. Therefore, further studies using in vivo digestibility in ruminant animals are required to confirm the ability of the protein extracted from indigofera to enhance rumen feed fermentation in low-protein basal diets. 

Aluminum (Al)-induced neurotoxicity is known to play a pivotal role in the development of various neurodegenerative diseases, and this is alleged to occur through neuroinflammation and oxidative stress in the brain. This study aimed to determine the effect of Ficus religiosa (FR) leaf extract on oxidative stress and neuroinflammation induced by Al exposure in the rat brain by estimating malondialdehyde (MDA), interleukin-6 (IL6), and total antioxidant (TAO) levels along with the degree of neurodegeneration in the brain of AlCl₃-administered and FR leaf extract-treated rats.  Two- to three-month-old male albino Wistar rats weighing 250–280 g were used in the present study. The animals were randomly divided into seven groups, with 12 rats in each group. The groups were categorized as control, Al-intoxicated, FR treatment groups of two dosages, FR control rats of two dosages, and FR pre-treatment group.  We observed a substantial increase in the levels of MDA and IL6 along with a decline in the TAO level in Al-intoxicated rats, suggesting increased lipid peroxidation (LPO), neuroinflammation, and oxidative stress, respectively. In the FR-treated animals, MDA as well as IL6 levels was decreased, and TAO was enhanced in addition to improved neuronal architecture, demonstrating the ameliorative effect of FR.  The present study observed a decline in LPO and neuroinflammation in FR-treated rats, demonstrating the protective effect of FR leaves against Al-induced neurotoxicity. The level of TAO also improved along with improvement in neuronal mass in FR-treated rats, adding to its ameliorative effect. However, further elaborate research is needed to confirm its therapeutic potential against inflammation-driven neurodegenerative diseases. 

Bovine interleukin 15 (bIL15) is a potential immunotherapy that can block the spread of bovine leukemia virus (BLV). However, immune checkpoints that maintain body homeostasis may reduce their effectiveness. Thus, an analysis of the effectiveness of bIL15 while blocking negative immune regulators is necessary. We aimed to obtain recombinant bIL15 (rbIL15) and determine its percentage using monoclonal antibodies against bovine cytotoxic T-lymphocyte antigen 4 (CTLA-4) and programmed cell death ligand 1 (PD-L1). To achieve this goal, peripheral blood mononuclear cells (PBMCs) from healthy and BLV+ cattle were treated with bIL15 using a CTLA-4– and PD-L1–blocking algorithm.  The codon-optimized bIL15 gene was synthesized under de novo conditions using polymerase chain reaction (PCR). The synthesized gene was cloned into pET28 and transformed into electrocompetent Escherichia coli BL21 cells; rbIL15 was purified using metal affinity chromatography and analyzed using sodium dodecyl-sulfate– polyacrylamide gel electrophoresis (SDS-PAGE) and western blotting. The expression of the Bcl2, STAT3, and STAT5 genes was studied using qualitative PCR. An enzyme-linked immunosorbent assay (ELISA) was used to analyze interferon (IFN)-γ production by rbIL15-treated mononuclear cells.  Analysis of rbIL15 using SDS-PAGE and western blotting revealed a specific product weighing 24 kDa. The optimal conditions for rbIL15 induction were 0.2 mM isopropyl-β-D-1-galactopyranoside and 37°C. When rbIL15 was added to PBMCs from healthy cattle, the Bcl2, STAT3, and STAT5 genes were expressed. ELISA of the culture medium of rbIL15-treated PBMCs revealed IFN-γ production. When PBMCs from healthy cows were treated with rbIL15, CTLA-4, and PD-L1 blockade together, they did not produce more IFN-γ than the rbIL15 group. Using PBMCs from BLV+ cattle, combination treatment increased IFN-γ production.  The biological activity of rbIL15 is characterized by the induction of transcription factors and the production of IFN-γ. Using rbIL15 with CTLA-4 and PD-L1 blockade in PBMCs from healthy and BLV+ cows led to the production of a transcription factor and cytokine. The results demonstrate the possibility of using this method to improve immunity and immunological memory in patients with chronic viral infections. 

Edible insects with high protein content and bioactive peptides with health promotion against chronic disease. Deficiency of nitric oxide (NO) contributes to hypertension, a leading cause of cardiovascular diseases and death worldwide. This study assessed the antihypertensive effects of house cricket protein hydrolysates (HCPH) in NO-deficient hypertensive rats. Male Sprague-Dawley rats (n = 12/group) were hypertensive after the administration of Nω-nitro-L-arginine methyl ester (L-NAME) at a dose of 50 mg/kg body weight (BW)/day in drinking water for 7 weeks. The animals were then treated with HCPH (250 or 500 mg/kg BW/day) or lisinopril (Lis) (1 mg/kg BW/day) for the last 4 weeks of L-NAME administration. Blood pressure (BP), vascular function, and structural changes, endothelial NO synthase (eNOS), and p47^phox nicotinamide adenine dinucleotide phosphate (NADPH) oxidase protein expression in aortic tissues, plasma nitrate/nitrite, plasma angiotensin-converting enzyme (ACE) activity, and oxidative stress markers in blood and tissues were evaluated.  Induction of hypertension resulted in significantly elevated BP, decreased plasma nitrate/nitrite concentration, abolished vascular function, and increased vascular wall thickness. Overproduction of carotid and mesenteric superoxide, increased plasma, heart, and kidney malondialdehyde, and protein carbonyl levels, and increased plasma ACE activity were observed. Down-expression of eNOS with overexpression of p47^phox NADPH oxidase subunit was also found in L-NAME hypertensive rats. Oral treatment with HCPH, particularly at a dose of 500 mg/kg BW/day, significantly alleviated these alterations in a manner comparable to that of Lis.  HCPH improved vascular function and exerted antihypertensive effects, mainly due to the improvement of NO bioavailability, reduction of oxidative stress, and inhibition of ACE. 

Lymphatic filariasis (LF) is a vector-borne disease in various regions of Indonesia. The transmission dynamics within a locality are intricately linked to the presence of the pathogen (microfilaria), definitive host (humans), intermediate host (mosquitoes), reservoir, and environmental factors. The geographic landscape of Central Bengkulu Regency, which is characterized by plantations, marshlands, and forests, serves as a suitable habitat for mosquitoes. Understanding species diversity, vector behaviors, habitat characteristics, and microfilarial presence is crucial for devising effective and efficient control strategies. This study aimed to identify species diversity, assess biting patterns, characterize larval habitats, and detect microfilarial presence in mosquitoes.  Mosquito collection was conducted using human landing collection (HLC) and resting collection indoors and outdoors for 6 months at a frequency of twice monthly from November 2022 to May 2023. The larvae were collected using dippers and pipettes. Adult mosquitoes and larvae were identified at the species level and analyzed using diversity indices. The measured larval bioecological parameters included physical, chemical, and biological conditions. The mosquito density obtained through HLC was calculated using the man-hour density (MHD) and man-biting rate (MBR) formulas. The presence of microfilaria was confirmed using a polymerase chain reaction.  A total of 808 adult mosquitoes from five genera and 18 species were captured, along with 485 larvae from four genera and eight species. The mosquito diversity was moderate. The dominant adult species included Armigeres subalbatus (44.8%), whereas Aedes albopictus (25.4%) and Ar. subalbatus (22.3%) were abundant larvae. The highest larval density was observed in natural ponds. The average MBR was three mosquitoes per person per night, with fluctuating nightly activity (mean MHD of 1.8 mosquitoes per person per hour). Larval habitats had temperatures of 25.4°C–28.7°C, illumination of 224–674 lx, and pH of 7.1–7.9, with over half being turbid and nearly two-thirds lacking predators. Microfilariae were not detected in the tested mosquitoes.  The presence of mosquitoes, their habitat, and the high density of Ar. subalbatus contributes to the transmission of LF in Central Bengkulu Regency, Indonesia. 

Diagnosis of trichinellosis at the intestinal stage during larval development is the primary challenge in the early detection and treatment of trichinellosis. The use of serine protease as a diagnostic marker for serological tests has been the subject of various studies, but data on Trichinella nativa serine protease in the intestinal phase are still insufficient for a proper diagnosis. This study aimed to establish the duration of the intestinal phase for early diagnosis and to determine the level of expression of the serine protease gene in T. nativa and Trichinella spiralis larvae.  We used European isolates from T. spiralis pigs and T. nativa larvae isolated from spontaneously infected wild carnivorous animals (wolf, Karaganda region) in Central Kazakhstan. Isolation of larvae from the meat of infected animals was carried out using the compressor method. For two species of Trichinella, 36 mice (in each group 18 mice) were infected with 250 larvae and euthanized by intramuscular injection of xylazine followed by an intravenous overdose of anestofol at 3, 5, 7, 14, 21, and 30 dpi (each day 3 infected mice) and one control group (3 mice). Sequencing and bioinformatics methods were used to determine the DNA and cDNA of the serine protease gene, and molecular methods (DNA extraction, reverse transcription polymerase chain reaction, and sequence) were used to measure the accumulation of serine protease transcripts in isolated larvae.  The results showed differences in the duration of intestinal phase between T. spiralis and T. nativa. The intestinal larvae of T. nativa were observed from 7 to 30 dpi, and the intensity of invasion increased up to 30 dpi (p < 0.001), while in the case of T. spiralis, the increase in larval growth in the intestinal phase decreased to 21 dpi, and only an increase of 1.6 ± 0.88 (p < 0.01) was detected at 30 dpi. T. nativa muscle larvae were detected at 21 dpi, compared with T. spiralis at 14 dpi. This characteristic was also reflected in the levels of serine protease transcripts in the samples. Accumulation was observed in both cases higher in the muscular stage of development, whereas the duration of the intestinal stage of T. nativa made it possible to detect serine protease at 30 dpi.  The intestinal stage of T. nativa lasts for 30 days, indicating that the use of T. nativa serine protease is useful for the identification of intestinal infection. Furthermore, this protein can be used to identify T. spiralis and T. nativa in laboratory samples. Serine protease can be used as a marker for serological diagnosis. Within the framework of the research topic, it is important to conduct further studies on the species specificity of the obtained recombinant protein. It is necessary to focus on identifying highly specific Trichinella proteins for early disease detection. 

Horses used for antisera production are repeatedly hyperimmunized to produce high levels of specific antibodies. This prolonged process can lead to various health issues, including amyloidosis, which involves the accumulation of amyloid proteins in organs and tissues, potentially causing organ dysfunction and failure. These horses are often retired when they no longer produce adequate antibody levels. This study aimed to evaluate the impact of prolonged antisera production on the health of retired horses by examining their blood biochemistry and serum amyloid A (SAA) levels, which are indicators of systemic inflammation and organ damage.  Blood samples were collected from 12 horses for this study. Nine horses were retired antisera-producing horses that had been discontinued for 2 years, while three healthy non-antisera-producing horses were used as controls. These twelve horses were divided into four groups based on the duration of their active period as antisera producers (never been used, 2–3 years, 4–5 years, and 6–7 years). We measured key blood biochemistry parameters and SAA levels to evaluate the health status of the horses.  Total protein, fibrinogen, and globulin levels were elevated, whereas other parameters remained normal. The findings indicate that despite normal SAA levels, the horses exhibited signs of ongoing health issues related to their previous use in antisera production, such as increased total plasma protein, fibrinogen, and globulin levels, as well as the presence of amyloid deposits in vital organs such as the liver and kidneys, as observed in post-mortem examinations.  Despite normal SAA levels, retired antisera-producing horses showed elevated total protein, fibrinogen, and globulin levels, indicating ongoing health issues. 

Pulmonary hypertension (PH) is a condition characterized by abnormally elevated pressure in the pulmonary vasculature. It is a common complication of myxomatous mitral valve disease (MMVD) in dogs. Several vasoactive substances, including endothelin-1 (ET-1), have been suggested to contribute to pathological changes in the pulmonary arteries of patients with PH. This study aimed to examine the local and systemic expression of ET-1 in dogs with PH secondary to MMVD.  Lung tissues were collected from 20 client-owned dogs during the first stage of the study and divided into three groups: normal dogs (n = 5), MMVD dogs (n = 8), and MMVD+PH dogs (n = 7). The expression of ET-1 and endothelin A receptor (ETAR) in the pulmonary arteries was determined using immunohistochemistry. Blood samples were collected from 61 client-owned dogs for the second stage of the study and divided into three groups: normal (n = 22), MMVD (n = 20), and MMVD+PH (n = 19). Plasma ET-1 concentration was measured using a sandwich enzyme-linked immunosorbent assay.  There was no difference in ET-1 and ETAR expression in the pulmonary arteries among the three groups. Similarly, there was no difference in the plasma ET-1 concentration between the groups. In addition, no correlation was found between the immunohistochemical expression of ET-1 and ETAR and the thickness of the pulmonary arteries or between the plasma ET-1 level and echocardiographic variables.  The lack of difference in the expression of ET-1 and ETAR in the pulmonary arteries and in the circulating ET-1 concentration among the studied groups suggests that ET-1 may not be related to the pathological development of PH secondary to MMVD in dogs. Due to the small sample size in this study, further research is needed to confirm these findings. 

During black cumin oil production, black cumin meal (BCM) is produced as a by-product. This study investigated the potential use of BCM to partly replace concentrate in a rice straw-based diet of fattening Garut lambs.  Twenty-eight heads of male Garut lambs aged approximately10 months with an average initial body weight of 20.7 kg/head (coefficient of variation 12.9%) were used. A completely randomized design was used to compare four different levels (0% [control], 5% [BCM-5], 10% [BCM-10], and 15% [BCM-15]) of BCM in rice straw- and concentrate-based diets on the performance of Garut lambs during 35- and 70-day feeding trials using seven replicates (n = 7). Blood metabolites and nutrient digestibility were also measured after the performance study.  BCM was rich in crude protein (36.8%) and tannins (21.6%). The BCM-10 and BCM-15 treatments increased (p < 0.05) average daily gain and feed efficiency compared with the control treatment in the 35-day feeding trial. All BCM treatments had greater dry matter (DM) intake compared with the Control at 70 days (p < 0.05). Furthermore, BCM-10 and BCM-15 had greater (p < 0.05) total protein, DM, and organic matter (OM) digestibility. BCM-15 had the highest (p < 0.01) blood triglyceride while BCM-10 tended to have higher (p < 0.1) blood glucose concentrations among the other treatments.  BCM supplementation can partly replace concentrate and improve the overall quality of rice straw and concentrate-based diets, resulting in improved performance of fattening Garut lambs due to increased DM and OM digestibility, as well as protein and energy absorption. Approximately 10% of the BCM supplementation is suggested as the optimum level. 

This study focuses on the scientific output of ehrlichiosis, a tick-borne disease that affects a variety of animal species, including dogs, cats, and livestock. Therefore, this study aimed to perform a scientometric mapping of the trends, impact, and thematic evolution of scientific production on ehrlichiosis in veterinary medicine.  The study design was descriptive and observational, with a quantitative scientometric approach. This study was based on Scopus data collection and analysis from 2018 to 2023. A literature search was conducted on February 12, 2024, and a total of 200 documents were found, of which 177 were articles, 15 book chapters, and eight reviews. A specific search formula was used to obtain documents. The documents were analyzed using SciVal and Bibliometrix in R Studio, focusing on four key metrics: Scholarly Output, View Count, Field-Weighted Citation Impact, and Citation Count.  This bibliometric study covered the period from 2018 to 2023 and analyzed 200 papers from 84 different sources. The average number of citations was 3595 and the mean age was 3.17 years. A total of 1874 keywords and 1085 authors were identified, with an average of 6.25 co-authors per paper. International co-authorship was present in 23% of the papers. The papers were distributed as articles (177), book chapters (15), and reviews (8).  The combination of these metrics enabled a more complete and accurate assessment of research performance. A total of 1874 keywords and 1085 authors were identified. The thematic evolution from “canine ehrlichiosis” and “Ehrlichia canis” to “dog” and “canine” was observed. Bradford’s and Lotka’s laws were confirmed, with some sources and authors generating most publications. 

Free-grazing duck (FGD) raising is a unique domestic duck production system that is widely practiced in several Asian countries, including Thailand. FGD is a significant reservoir for influenza A viruses (IAVs). In this study, we genetically characterized IAV-H10N6 and IAV-H10N7 isolated from avian influenza surveillance in FGDs in Thailand.  We collected 640 swab samples from 29 FGD flocks located in 6 provinces of Thailand. IAVs were isolated from swab samples using egg inoculation. Hemagglutination test-positive samples were then subjected to IAV detection. Viral RNA was subjected to IAV detection using real-time reverse-transcription polymerase chain reaction (rRT-PCR) specific to matrix (M) gene. IAV subtypes were identified using the RT-PCR assay specific to all hemagglutinin and neuraminidase subtypes. Whole-genome sequencing of IAVs was performed to genetically characterize IAV-H10N6 and IAV-H10N7.  Our results showed that 41 (6.41%) samples tested positive for IAV using rRT-PCR specific to the M gene. Among these, only two IAVs were subtypes as IAV-H10N6 and IAV-H10N7 and were subjected to whole-genome sequencing. IAV-H10N6 and IAV-H10N7 belonged to the Eurasian lineage and did not show any evidence of reassortment from the North American lineage. The viruses exhibited low-pathogenic characteristics and preferred binding to avian-type receptors. Genetic analysis revealed no mutations in PB2 and M genes, unlike human IAV-H10N3 and IAV-H10N8, which exhibited increased virulence in mammals. IAV-H10N6 and IAV-H10N7 viruses have less potential as zoonotic viruses. However, IAV in FGDs should be monitored for novel reassortant or zoonotic viruses. This study provides information on the genetic characteristics and diversity of IAV-H10N6 and IAV-H10N7 that are circulated in FGDs in Thailand. 

Immunoglobulin (Ig)Y, a specific type of Ig found in chicken eggs, has potential use in the diagnosis of human and animal diseases. This study assessed the feasibility of using spent laying hens to produce IgY. In addition, the effects of antigen injection on egg and antibody production in new and spent laying hens were compared.  Hens were intramuscularly injected with three booster shots of antigens. IgY was extracted from egg yolks using polyethylene glycol 6000 precipitation followed by dialysis.  Spent laying hens (83 weeks) consistently showed lower egg production than new laying hens (27 weeks) throughout the study. Post-immunization, a further decline in egg production was observed in spent laying hens, and egg production stopped after the second antigen injection. Eggs from spent laying hens were less dense than eggs from new hens. Despite lower IgY levels in eggs from spent laying hens, the heavy-to-light chain ratio remained consistent in both groups. Notably, IgY from spent and new laying hens demonstrated effective hemagglutination against cat erythrocytes in the A blood group.  This study demonstrated the potential of using spent laying hens to produce IgY, with significant implications for future research, immunotherapy, and diagnostic applications, despite the observed reduction in egg production compared with new laying hens. 

Pig industries are currently facing a crisis in terms of protein and energy costs. Proteases were used to increase protein digestibility and metabolizable energy (ME) in diets. This study evaluated the effects of protease supplementation on in vitro protein digestibility and productive performance in starter-to-finisher pigs.  A total of 691 starter pigs were randomly allocated into three dietary treatments using a randomized complete block design. Diets were provided in three phases according to body weight (BW): Starter, grower, and finisher phases. Each phase was fed for 30, 60, and 24 days of treatment diets as T1: basal diet and T2 and T3: the basal diet supplemented with 240 ppm protease reduced by 50 kcal/kg ME plus 1% crude protein (CP) and by 100 kcal/kg ME plus 2% CP, respectively. Protease and in vitro protein digestibility were measured. BW and feed intake were recorded to calculate the average daily gain (ADG), average daily feed intake (ADFI), feed-to-gain (F:G), and gain-to-feed (G:F) ratios.  There were no significant differences (p > 0.05) in the percentage of in vitro protein digestibility between the groups with and without protease supplementation. In the finisher phase, T2 had lower (p < 0.05) ADFI and F:G than T1 and T3. Overall, T3 had lower (p < 0.05) ADG, ADFI, and F:G than T1 and T2.  Protease supplementation significantly affects protein digestibility. Supplementing basal diets with 240 ppm protease reduced ME by 50 kcal/kg and CP by 1% without affecting ADG, ADFI, F:G, and G:F ratios for starter-to-finisher pigs.