Pigs are the main livestock commodity in Bali Province, Indonesia, where traditional farming practices are widely used. Traditional pig farmers are often closely associated with poverty and a perceived lack of knowledge regarding health and hygiene. Data on soil-transmitted helminthiasis (STH) and risk factors associated with STH worm infection among traditional pig farmers in Bali were previously unavailable. This study aimed to analyze the prevalence and risk factors for STH infections among traditional pig farmers in Bali Province, Indonesia. This study involved the fecal examination of 238 traditional pig farmers from all areas of Bali Province in Indonesia. In addition, several pig feces samples were combined into one pooled sample belonging to each farm. All fresh fecal samples were stored in a 5% formaldehyde solution before being analyzed using concentration flotation techniques. Subsequently, risk factors were determined through an interview and a questionnaire. The odds ratio (OR) and Chi-square tests were used to determine the risk factors associated with STH infections. The result showed that there was a high prevalence of STH infections among traditional pig farmers and pig farms in Bali, with rates of 21.8% and 76.5%, respectively. This could be due to risk factors, such as personal hygiene (OR: 5.756; confidence interval [CI]: 2.96-11.193; p=0.00), sanitation (OR: 1.914; CI: 1.024-3.576; p=0.042), education level (OR: 7.579; CI: 2.621-21.915; p=0.00), household income (OR: 2.447; CI: 1.122-5.338; p=0.025), and occupation (OR: 2.95; CI: 1.356-6.415; p=0.006). The infections seen in farmers were distributed among hookworm, Ascaris spp., and Trichuris spp., at 15.1%, 9.2%, and 4.2%, respectively. The risk factors associated with infections of STH and Ascaris spp. were personal hygiene, home sanitation, education level, household income, and having a primary occupation as a traditional pig farmer. In contrast, personal hygiene, education level, and primary occupation were the only risk factors for hookworm infection, while personal hygiene and home sanitation were the risk factors associated with Trichuris spp. infection. The limitation of this study was that the number of samples was relatively small due to the difficulty of obtaining stool samples from traditional pig farmers, with many individuals refusing to provide their stool for inspection. We suggest that future research focus on identifying the species of worms that infect traditional pig farmers and to better identify the zoonotic link of STH transmission from pigs to humans. Keywords: Ascaris, hookworm, risk factor, soil-transmitted helminthiasis, traditional pig farmer, Trichuris.

Exosome-derived microRNA (miRNA) has been widely studied as a non-invasive candidate biomarker for tumor diagnosis in humans and dogs. Its application, however, was primarily focused on intraspecies usage for individual tumor type diagnosis. This study aimed to gain insight into its application as a cross-species differential tumor diagnostic tool; we demonstrated the process of identifying and using exosome-derived miRNA as biomarkers for the classification of lymphoid and mammary tumor cell lines in humans and dogs. Exosome-derived miRNA sequencing data from B-cell lymphoid tumor cell lines (n=13), mammary tumor cell lines (n=8), and normal mammary epithelium cultures (n=4) were pre-processed in humans and dogs. F-test and rank product (RP) analyses were used to select candidate miRNA orthologs for tumor cell line classification. The classification was carried out using an optimized support vector machine (SVM) with various kernel classifiers, including linear SVM, polynomial SVM, and radial basis function SVM. The receiver operating characteristic and precision-recall curves were used to assess the performance of all models. MIR10B, MIR21, and MIR30E were chosen as the candidate orthologs from a total of 236 human-dog miRNA orthologs (p≤0.01, F-test score ≥10, and RP score ≤10). Their use of polynomial SVM provided the best performance in classifying samples from various tumor cell lines and normal epithelial culture. The study successfully demonstrated a method for identifying and utilizing candidate human-dog exosome-derived miRNA orthologs for differential tumor cell line classification. Such findings shed light on a novel non-invasive tumor diagnostic tool that could be used in both human and veterinary medicine in the future. Keywords: exosome-derived microRNA, meta-analysis, ortholog, support vector machine, tumor.

Salmonella is a major foodborne pathogen in the poultry industry, wherein the control measures may include sanitation and antibacterial and vaccines. However, there have been severe global restrictions on using anti-Salmonella antibacterial agents in livestock. This situation, along with rapidly increasing drug-resistant bacterial species, has led to the exploration of unconventional methods to control Salmonella infection in poultry. In recent years, selection techniques of promising DNA aptamers have begun to permeate several medical branches, resulting in the development of numerous anti-Salmonella DNA aptamers, most of which are used as sensing molecules for diagnostic purposes. These DNA aptamers have been demonstrated to interfere with bacterial growth, multiplication, and viability. Aptamers formed in rolling circle amplification products (RCA-p) could improve the potential action of aptamer interference with bacteria. This study aimed to test the use of single-stranded DNA aptamers in the form of RCA-p as a bacteriostatic to Salmonella in vitro. Salmonella Typhimurium and Salmonella Enteritidis isolates were subjected to the action of anti-ST and anti-SE DNA aptamers in the form of RCA-p. Each isolate was grown on MacConkey and Luria-Bertani agar media separately in different concentrations in the presence or absence of the cognate RCA-p. The anti-Salmonella species DNA aptamer-based RCA-p were capable of reducing bacterial growth to significant levels in vitro. We describe a potential solution for the rapidly developing drug resistance of several bacterial species. Our findings suggested that the use of non-toxic, non-immunogenic, and low-cost DNA aptamers targeting Salmonella in the form of RCA-p could inhibit the bacterial growth rate. Unlike polymerase chain reaction, RCA yields tandem repeats of single-stranded DNA at isothermal conditions, which would increase the probability of receptor-ligand clustering and increase affinity. Furthermore, as our RCA template was bivalent with two DNA aptamer sequences, we could target multiple sites or antigens on a bacterial cell. Keywords: DNA aptamers, rolling circle amplification, Salmonella.

Breast cancer is the most frequent malignancy in women. The consumption of phytochemical components from plants may play an essential role in preventing and treating this cancer. This study aimed to investigate the anti-cancer activity of an ethanolic extract of red okra pods (EEROP) in rats (Rattus norvegicus) induced by N-methyl-N-nitrosourea (MNU). The experimental animals were divided into six groups (n=5/group), namely, KN (normal control, without any treatment), K– (negative control, exposed to MNU without EEROP), K+ (positive control, exposed to MNU and Methotrexate), and the treatment Groups P1, P2, and P3 (exposed to MNU and EEROP at doses of 50, 100, and 200 mg/kg body weight [BW], respectively). Intraperitoneal delivery of MNU and EEROP oral administration was carried out for 8 weeks. After the end of treatment, the parameters of cytokines, CD4+ and CD8+ T cells, and mammary gland histology were measured. The results showed that EEROP at doses of 100 and 200 mg/kg BW significantly downregulated interleukin (IL)-6, IL-1β, tumor necrosis factor (TNF)-α, IL-17, IL-10, and tumor growth factor-β (p<0.05). In addition, doses of 200 mg/kg BW significantly increased the activity of CD4+ and CD8+ T cells, prevented the proliferation of mammary gland epithelial cells, and yielded a significantly thinner epithelium of the mammary gland (p<0.05). It can be concluded that EEROP was an effective anti-cancer agent by modulating the immune response. Further studies using a nanoparticle system are warranted to achieve optimal working conditions for these bioactive compounds. Keywords: cancer, cytokines, histology of mammary gland, N-methyl-N-nitrosourea, red okra pods.

Foodborne illnesses are a serious challenge to human health and the economic sector. For example, salmonellosis remains a burden in developed and developing nations. Rapid and reliable molecular methods to identify Salmonella strains are essential for minimizing human infection. This study aimed to identify Salmonella spp. in raw milk and dairy products using conventional and molecular techniques and to test the antibiotic susceptibility of the isolated strains. One hundred and thirty-one milk and dairy product samples were randomly collected from different localities in Libya. Samples were examined for the presence of Salmonella by conventional culture techniques, including cultivation in Rappaport-Vassiliadis broth and streaking on xylose lysine deoxycholate agar. Identification also used polymerase chain reaction and partial sequencing of 16S rDNA. Twenty-four antibiotics were used for the examination of antimicrobial resistance of Salmonella spp. isolates with the agar disk diffusion method (Kirby–Bauer technique). Multi-antibiotic resistance index and antibiotic resistance index (ARI)for Salmonella enterica isolates were calculated. Twenty-one of 131 samples (16%) were positive for Salmonella spp. recovered from 9 (16%), 2 (11%), 4 (22.2%), and 6 (46%) samples of raw cow milk, fermented raw milk, and fresh locally made soft cheeses, Maasora and Ricotta), respectively. Samples of ice cream, milk powder, and infant formula showed no Salmonella spp. contamination. Only 9 of 21 (42.8%) isolates were confirmed as S. enterica by partial sequence 16S rDNA analysis. All isolates were resistant to amoxycillin, bacitracin, penicillin G, lincomycin, vancomycin, clindamycin, and cloxacillin with an ARI of 0.042. In contrast, all tested strains were sensitive to levofloxacin, doxycycline, and ciprofloxacin. In addition, all of the tested isolates (100%) were resistant to more than one antibiotic. This study demonstrated the applicability of molecular techniques, compared with conventional methods, as preferable for the identification of Salmonella in milk and dairy products and thus reduction of milk-borne transmission to the consumers. From the view of public health, isolation and identification of Salmonella multidrug-resistant strains from raw cow's milk and locally prepared dairy products sold in the Libyan markets indicate the need to improve the handling and processing of milk and dairy products to minimize the prevalence of Salmonella, one of the most important foodborne microorganisms that cause food poisoning. Keywords: 16S rDNA partial sequence, antibiotic resistance, dairy products, Libya, milk, Salmonella spp.

Fresh produce farms represents a major source of concern since they are becoming increasingly antibiotic resistant. This study aimed to investigate t he occurrence of carbapenemase and extended-spectrum-beta-lactamases (ESBL) - producing genes in Klebsiella pneumoniae isolated from fresh produce farms in Egypt, irrigation water, and people working in these fields. One hundred tomatoes from typical farms were collected in plastic bags. The study also included 20 surface-water samples from different irrigation watersheds in fresh produce farms, as well as 50 feces samples from farmworkers. Suspected K. pneumoniae was grown on Eosin Methylene Blue agar for 24 h before being biochemically identified using the RapID ONE test. PCR was used to detect carbapenemase (blaKPC, blaOXA-48, and blaNDM) and ESBL (blaSHV, blaTEM, and blaCTX) expressing genes on isolates. K. pneumoniae was identified in 30% of water and 10% of worker samples, while only one isolate was found in tomato samples. One of the six irrigation water isolates tested positive for carbapenem-resistant NDM. In contrast, two isolates tested positive for ESBL determinants, one of which was blaSHV and the other having both blaSHV and blaTEM genes. Two of the five K. pneumoniae isolates from farmworkers were positive for blaNDM, with one isolate also testing positive for blaSHV and blaTEM. The blaOXA-48 gene was also discovered in the carbapenem-resistant K. pneumoniae tomato isolate used in this study. Carbapenemase- and ESBL-producing K. pneumoniae were found in fresh produce farms, implying that these resistance genes were being passed down to Egyptian consumers. Keywords: carbapenemase, extended-spectrum-beta-lactamases, fresh produce, humans, irrigation water, Klebsiella pneumonia.

Basil is well known as a medicinal plant that contains high essential oils and antioxidant compounds that have the potential to improve ovarian development. Thus, basil may have the potential to improve the growth and development of the uterus and placenta for optimal prenatal growth of offspring. This study aimed to evaluate the effect of Indonesian basil maceration on gonad development of mature female albino rats. Fifteen 8-week-old female Sprague-Dawley rats, at the diestrus stage of the estrus cycle, were divided into three different treatment groups: Control group (mineral water), bas-low group (1% of basil maceration), and bas-high group (5% of basil maceration). Basil maceration was dissolved and administered in mineral drinking water, and the treatments were given for 20 days (4 estrus cycles). At the end of the treatment period, serum follicle-stimulating hormone (FSH), estradiol, and progesterone (Pg) were measured using enzyme-linked immunosorbent assay. The relative weight of the ovary and uterus; diameter and length of uterine cornual; vascularization of uterus; the diameter of uterine glands; the number of primary, secondary, and tertiary de Graaf follicles; the number of corpora luteum; as well as the expression of vascular endothelial growth factor (VEGF) in the ovary were determined. There was no significant difference (p>0.05) in the serum FSH level of rats treated with basil maceration drinking water doses of 1% and 5% compared to the control group. However, serum estradiol and Pg concentrations in the 1% and 5% basil maceration groups were significantly higher (p<0.05) than those of the control group. Furthermore, 1% and 5% basil maceration significantly increased the uterus's relative weight, diameter, and vascularization. Serum estradiol concentrations contributed to the elevated expression of VEGF compared to Pg. Administration of basil maceration for 20 days before mating could improve follicle growth and development, eventually increasing estradiol synthesis and secretion, thus improving the uterus's preparation for implantation. This makes basil maceration an attractive candidate in clinical research to enhance the growth and development of the uterus and placenta, which will better support the optimum prenatal growth and development of embryos and fetuses, resulting in superior offspring. Keywords: Indonesian basil, rat, reproductive system.

Roundworms cause infections in the avian population that lead to illness and poor production. The singing zebra dove is an economically important animal in the Indo-Malay region. The prevalence of these parasitic groups in zebra doves is unknown. This study estimated the prevalence and associated human risk factors of gastrointestinal nematode infections in zebra dove farming. A cross-sectional survey was conducted from January to April 2021. The study was conducted on 184 doves in three zebra dove farms. Fecal samples were collected from pooled zebra dove droppings. Major proportions and infection intensity of gastrointestinal nematodes were morphologically identified and morphometrically investigated. Associated human factors were assessed through the interview surveys among farmers. Results showed that 36.96% of the zebra doves were infected. The primary nematodes were Ascaridia galli (34.78%), Heterakis gallinarum (6.52%), Trichostrongylus tenuis (2.17%), Syngamus spp. (4.35%), and Amidostomum spp. (2.17%). The primary human factors that contribute to parasitic infection were poor hygiene, food contamination with parasites, and inappropriate deworming. There was a high prevalence of gastrointestinal nematodes in the zebra dove in the close cage system. Human factors played key roles as risk factors, and improves farming management will help reduce parasitic infections. However, these nematodes may contribute to poor health status and poor productivity of zebra doves. Further extensive studies on clinical signs and pathological changes should be conducted. Keywords: Ascaridia galli, gastrointestinal nematodes, Geopelia striata, human factors, Syngamus spp., zebra dove.

Salmonellosis is one of the most common foodborne diseases in the world, and the increasing antibiotic resistance in Salmonella enterica subsp. enterica recovered from food animals constitutes an important risk from a One Health approach. This study aimed to characterize antibiotic resistance and some of its associated resistance determinants in different S. enterica serovars isolated from pigs in Argentina. A retrospective study was conducted on Salmonella strains isolated between 2011 and 2015 from pigs in the Pampean region of Argentina. The antimicrobial susceptibility patterns to 21 antimicrobials and some antibiotic resistance determinants were characterized in 55 Salmonella isolates, representing 58 farms. We identified 56% (n=30) of the strains as multidrug-resistant, where resistance to tetracycline (62%, n=34), ampicillin (53%, n=29), nalidixic acid (53%, n=29), chloramphenicol (33%, n=18), and trimethoprim-sulfamethoxazole (31%, n=17) was most common. The wide range of resistance to ampicillin correlates with the presence of TEM type β-lactamases in the strains. However, high susceptibility was found in the new generation of β-lactams. Fluoroquinolone resistance is a major concern. Most strains with decreased susceptibility to ciprofloxacin showed gyrA mutations and plasmid-mediated quinolone resistance gene qnrB. Here, we identified broad resistance to some antibiotics frequently used in human therapeutics and several easily transferable resistance mechanisms that could endanger public health. Keywords: environment, health, livestock, salmonellosis.

Ovine anaplasmosis is a rickettsial disease caused by Anaplasma spp. These Gram-negative intracellular bacteria are mainly transmitted by ticks and infected blood cells of caprine, ovine, and wild small ruminants. At present, epidemiological data on anaplasmosis in cattle, dogs, and camels in Egypt are available, but the data about Anaplasma spp. in sheep and goat are scarce. This study aimed to determine the seroprevalence of Anaplasma spp. in small ruminants and assess the associated risk factors. A cross-sectional study was performed to investigate the seroprevalence of Anaplasma spp. in 300 sheep and 300 goats from four governorates in North Egypt using a commercial competitive enzyme-linked immunosorbent assays kit, and the associated risk factors for the infection were evaluated. Overall, the seroprevalence of anti-Anaplasma antibodies was 18.3% and 21.3% in sheep and goats, respectively. A multivariable logistic regression model was used to determine the association between risk factors and Anaplasma spp. infection. Age, animal husbandry, acaricide use, tick infestation, and contact with cattle were the primary risk factors for Anaplasma seropositivity. This study confirms the presence of antibodies against Anaplasma spp. in small ruminants from Egypt. This is the first study to assess the associated risk factors for Anaplasma infection in small ruminants from Egypt. Further studies are needed to improve the understanding of the associated disease factors, facilitating the development of new procedures for control of anaplasmosis in livestock. Keywords: Anaplasma spp., competitive enzyme-linked immunosorbent assay, Egypt, risk factors, small ruminants.

Dairy cattle breeding plays a significant role in providing the population with high-quality, reasonably priced goods. The development of this industry and its effectiveness depends on the proper use of available feed products. Feed additives (FAs), as a rule, should compensate for missing elements in the diet. This study aimed to determine the effect of the FA Vermikom on blood parameters, as well as milk physicochemical and mineral composition and yield in lactating dairy cows. A total of 30 Holstein cows, with an average weight of 650±5 kg, were randomly divided into three groups of 10. Over a period of 5 months, each group was fed one of three diets: Mixed main diet without supplements (control), main diet supplemented with 2% Vermikom, and main diet supplemented with 4% Vermikom. Hematological parameters, productivity, and physicochemical and mineral composition of milk from animals provided the FA Vermikom were higher than those of the control group. In the Vermikom groups, the hemoglobin content was higher by 5.75%, calcium by an average of 10.8%, and total protein by 2.5%. The average daily milk yield was also higher by 8.4% than the control group. Regarding the mineral composition of the milk, the content of calcium, phosphorus, and iron exceeded that of the control on average by 2.9%, 3.4%, and 14.8%, respectively. We propose introducing 4% Vermikom into the diet of dairy cows based on the study results. Future research will expand our knowledge regarding cows' needs for all recommended nutrients, thereby improving animal productivity and milk quality. The results obtained contribute to further expanding the food base of animal husbandry in the Republic of Kazakhstan. Keywords: diet, feed additive, hematology, milk productivity, milk quality, vermiculite.

Rabies remains a public health concern in Indonesia, and the coronavirus disease (COVID-19) pandemic has stymied rabies prevention and control efforts. There is a need to transform the rabies program to be adaptable to pandemic situations to improve program coverage on dog vaccination and rabies surveillance. This study aimed to create a rabies control (RaCon) mobile application for a community-based rabies surveillance system during COVID-19 in Bali, Indonesia. We employ the Design Science Research methodology. Surveillance officers, veterinarians, community leaders, outreach workers, and dog owners participated in a series of offline in-depth interviews and focus group discussions. The RaCon prototype was evaluated using the Post-Study System Usability Questionnaire (PSSUQ) framework, which included the system's usefulness, information quality, and interface quality. In this study, we used both a qualitative (n=50) and quantitative (n=342) approach. According to the findings of this study, integrating public health and animal health into the rabies surveillance system are critical to supporting the One Health approach and encouraging community engagement in rabies programs. The RaCon prototype is expected to include features such as pet ownership, case report, news and announcements, nearest vet, health information, outbreak radar, emergency call, and app feedback. The RaCon prototype passed both qualitative and quantitative evaluations, indicating that it could be used to support the rabies surveillance system, particularly in the COVID-19 situation. The RaCon prototype was accepted by the users and got positive feedback in terms of the system's usefulness, information quality, and interface quality dimension. As a result, this prototype has the potential to be integrated into the rabies surveillance system in Bali, particularly to strengthen the community-based rabies surveillance system. Even though this prototype received positive feedback, this study focuses solely on the design development and evaluation of its user interface. As a result, further development is required before incorporating RaCon into the rabies prevention and control program. Keywords: community-based intervention, One Health, post-study system usability questionnaire, rabies, surveillance system.

As the number of wild Asian elephants (Elephas maximus) continues to decline, maintaining healthy populations under human care is vital. Male fertility assessment is essential for understanding the reproductive status, which can help to uncover underlying problems and improve the rate of pregnancy success. The objectives of this study in Asian elephants were as follows: (1) To investigate the semen characteristics; (2) to compare the relative seminal vesicle size and semen characteristics; (3) to compare the semen characteristics between good-motile (>60% progressive motility) and poor-motile (<60% progressive motility) ejaculates; and (4) to investigate the pregnancy success rate after artificial insemination (AI) with combined chilled and frozen semen. In total, 153 ejaculates were collected by manual rectal stimulation from 25 bulls. The volume, pH, sperm concentration, progressive motility, viability, morphology, and membrane integrity were investigated in each ejaculate. Assessment of accessory sex glands was conducted using transrectal ultrasonography to compare the relative seminal vesicle size and semen characteristics, and the bulls were divided into two groups according to the size of the ampulla (<7 or ≥7 cm2). For the comparison of good and poor-motile ejaculates and semen characteristics, the samples were divided into two groups: Good-motile (>60% progressive motility) and poor-motile (<60% progressive motility) ejaculates. Semen ejaculates for AI were collected from three bulls. The estrous cycles of four females were monitored using an enzyme immunoassay. Seven AI attempts were conducted using frozen and/or chilled semen by endoscopic visualization. AI was repeated 1 day before the luteinizing hormone surge, on the day of the surge, and 1 day after the surge. Pregnancy was confirmed by monitoring the serum progesterone profile and the abdomen and mammary glands changes. From 153 ejaculates, the mean±standard error values of progressive motility, semen volume, sperm concentration, pH, and viability were 40.18%±2.28%, 40.94±3.86 mL, 1,205.58±62.26×106 sperm/mL, 7.50±0.10, and 56.17%±1.96%, respectively. Comparing ampulla size and semen characteristics revealed that the bulls with ampullae of ≥7 cm2 yielded significantly larger volume ejaculates. However, there were no significant differences in sperm motility and concentration. The comparison of semen characteristics between good- and poor-motile ejaculates revealed that the former had significantly higher pH, viability, normal acrosomes, intact membranes, and normal head and tail morphology but often had a significantly lower volume and sperm concentration. From seven AI attempts in four females, one female had a confirmed pregnancy (14.3% pregnancy rate), and delivered a healthy live female baby weighing 128 kg at 21 months and 12 days of gestation. The baby is now 3 years old and in a healthy condition, with normally developing growth and behavior. The semen characteristics of Asian elephants can be used as the baseline reference for further applications. The ampullae size indicates semen quantity but not quality. Our success in producing an elephant calf from AI using frozen and chilled semen demonstrated that AI can be used as an alternative approach for the breeding management of Asian elephants. However, the semen of Asian elephants is of poor quality, especially in terms of membrane integrity; thus, the improvement in semen quality through intensive and careful management of elephant health and fertility remains a challenge for the future. Furthermore, a sperm bank should be established to develop sperm cryopreservation, which will be invaluable for improving the genetic diversity of the Asian elephant. Keywords: artificial insemination, Asian elephant, hormone, semen quality, sex accessory glands.

Colostrum composition and importance for newborn organisms were repeatedly studied. However, the interest in transitional milk usefulness is weak and recommendations concerning transition milk intake are not developed. The aim of this study was to evaluate whether transition milk intake after colostrum consumption affects the chances of calf infection with Cryptosporidium spp. We collected data for Cryptosporidium spp. infection from calves (n=425) divided into three groups: The first group – supervised colostrum and transition milk intake; the second group – supervised colostrum and whole milk intake; and the third group – not supervised colostrum and whole milk intake. To detect oocysts of Cryptosporidium spp. in feces, the flotation method was used, and slides were stained using the modified Ziehl-Neelsen method. Generalized linear mixed modeling was conducted to determine whether the explanatory variable – the management of colostrum and transition milk feeding with three categories (three research groups) – was related to the probability of calves incurring infection with Cryptosporidium spp. In the first group, 26.1% of calves were positive for the presence of Cryptosporidium spp. oocysts, in the second – 37.2%, and in the third – 44.1%. Statistical data analysis showed that calves who did not receive transition milk after colostrum consumption had increased chances of having Cryptosporidium spp. (by 1.90-2.47 times on average). The main results showed that the management of colostrum and transition milk feeding is related to Cryptosporidium spp. infection, indicating that both colostrum and transitional milk play a significant role in controlling pathogenic infections. The most effective management of colostrum and transition milk feeding against Cryptosporidium spp. infection is the timely intake of an adequate amount of colostrum followed by transitional milk consumption for at least 2 weeks before weaning from the dam. Keywords: calves, colostrum, Cryptosporidium, neonatal diarrhea, transition milk.

Vaccines are one of the important tools for fighting diseases and limiting their spread. The development of vaccines with high efficacy against diseases is essential. Ionizing radiation is the method used for the preparation of the irradiated gamma Mannheimia haemolytica vaccine. The study aimed to measure the metabolic activity and electron microscopic examination of the irradiated bacterial cells and immunological efficiency of different preparations of the irradiated M. haemolytica vaccine. The irradiated vaccines were prepared in three forms at a dose of 2×109 colony-forming unit (CFU) (irradiated M. haemolytica, trehalose irradiated M. haemolytica, and trehalose lyophilized irradiated M. haemolytica). The formalin-killed vaccine was prepared at a dose of 2×109 CFU. Scanning electron microscopy was used to determine the difference between the non-irradiated bacterial cells and the bacterial cells exposed to gamma radiation. The metabolic activity of the irradiated bacterial cells was measured using the Alamar blue technique. Rabbits were divided into five groups (control, vaccinated groups with the formalin-killed vaccine, irradiated bacterial cells without trehalose, trehalose irradiated bacteria, and trehalose lyophilized irradiated bacterial cells). The rabbits were subcutaneously inoculated twice in 2-week intervals. Enzyme-linked immunosorbent assay, interferon-gamma (IFNγ), and interleukin 4 (IL4) assays were used to evaluate the vaccines' immunological efficiency in rabbits. The metabolic activity tests showed that the bacterial cells exposed to gamma radiation at the lowest lethal dose have metabolic activity. The difference in the metabolic activity between preparations of the irradiated bacterial cells varied according to the cell concentration and incubation time. The highest level of metabolic activity was 8 h after incubation in the nutrient broth medium compared with 4 and 18 h. The scanning electron microscopy of irradiated bacterial cells showed a cavity at the bacterial cell center without rupture of the surrounding cell membrane compared to the non-irradiated bacterial cells. The antibody level in the groups vaccinated with the different preparations of the irradiated bacterial cells was high compared with the control and formalin-killed vaccine groups. The level of the IFNγ showed an increase after the second dose in the group vaccinated with irradiated bacterial cells without trehalose compared with the other groups. The IL4 level in the vaccinated groups with the irradiated bacterial cells without trehalose, irradiated bacterial cells with trehalose, and trehalose lyophilized irradiated bacterial cells were at a high level when compared with the formalin-killed vaccinated group and control group after the second inoculation. The irradiated M. haemolytica vaccine provides a wide range of humoral and cellular immunity. This study showed high immunological efficiency in rabbits inoculated with the irradiated M. haemolytica vaccine that was shown in the high levels of antibodies (IFNγ and IL4) compared with the group treated with the formalin-killed vaccine. The second dose of irradiated M. haemolytica vaccine is an immune booster that gives the irradiated vaccine a long-acting immunological efficiency. Keywords: enzyme-linked immunosorbent assay, gamma radiation, interferon-gamma, interleukin 4, Mannheimia haemolytica, vaccine.

The need for fish seeds resistant to bacterial and viral infections has encouraged studies on the molecular pathogenesis mechanism of Vibrio bacteria, such as Vibrio alginolyticus and Vibrio harveyi, regarding the receptor organs, protein adhesion mechanisms, and antibody responses of the humpback grouper. This study aims to confirm the characteristics of the specific proteins expressed in the receptor organ of the humpback grouper (Cromileptes altivelis) using the expression of V. alginolyticus and V. harveyi bacteria. The study was conducted by isolating crude protein and whole cells from both the Vibrio bacteria. In addition, serum and organ tissue were also isolated from fish samples. Then, hemagglutination and dot blot tests with polyacrylamide gel electrophoresis analysis were performed to determine the highest expression of receptor from the whole bacterial cells and crude protein from both healthy and infected (V. alginolyticus and V. harveyi) fishes. Scanning electron microscope results showed that V. alginolyticus and V. harveyi could express bundle-forming pili, which is involved in bacterial autoaggregation and the mediation of the initial attachment of bacteria to their host cells. These results indicated that all the specific receptors for protein in fish organs recognized vibriosis antigens. The specificity test showed that the brain, eye, and kidney organs' receptors provided a quality and quantity level of responses at 22.63, 53.95, and 43.15 kDa, respectively. The polyclonal anti-V. alginolyticus immunoglobulin M (IgM) antibodies were more cross-reactive than the anti-V. harveyi IgM. Hence, this shows that V. alginolyticus bacteria are more pathogenic than V. harveyi. In the future, the molecular characteristics of V. alginolyticus and V. harveyi antigens and the specific receptor organ proteins in the humpback grouper can be developed as the basis for constructing molecular peptide-based vaccine materials. Keywords: crude protein, humpback grouper, Vibrio alginolyticus, Vibrio harveyi, western blot.

New substances for neoplasm treatment have to be carefully studied to minimize adverse effects and prevent disease progression stimulation. Jatobá is a typical tree of the Cerrado and Caatinga biome, with antifungal, antimicrobial, larvicide, antioxidant, and antiproliferative properties. This study aimed to investigate the action of the crude extract of Jatobá leaves (EBFJ) on canine osteosarcoma (CO) cells and analyze the expression of biomarkers in neoplasm progression. D17 cells were cultured and subjected to treatment with EBFJ at different concentrations (10 μg/ mL; 100 μg/mL; 1000 μg/mL; 2000 μg/mL; and 5000 μg/mL) and exposure times (24 h, 48 h, and 72 h). The tetrazolium reduction assay and the immunocytochemistry technique, with anti-Bcl2, anti-p53, and anti-Ki-67 antibodies, were used to observe the effect of the extract on cell proliferation. Doses of 2000 μg and 5000 μg had cell viability of 300.80% and 361.84%, respectively. The extract did not show significant cytotoxicity of samples with the control group. The confluence of cells, the number of labeled cells, and the expression of Bcl2, Ki-67, and p53 were higher in the groups treated with EBFJ, with a statistical difference from the group without treatment. EBFJ was not cytotoxic and had a proliferative effect on CO D17 cells. The confluence of cells, the number of labeled cells, and the expression of Bcl2, Ki-67, and p53 were higher in the groups treated with the extract. Keywords: antineoplastic chemotherapy, biodiversity, bone tumors, Hymenaea martiana Hayne, medicinal plants.

Bovine viral diarrhea virus (BVDV) and classical swine fever virus (CSFV) are important pathogens of cattle and pigs, respectively, and belong to the genus Pestivirus. As CSFV has been shown to infect cattle, it can create diagnostic challenges of BVDV results through possible cross-reactivity where cattle could be exposed to pigs and CSFV. This study aimed to determine the possible cross-reactivity of BVDV and CSFV enzyme-linked immunosorbent assay (ELISA) results for antigen (Ag) and antibody (Ab) among smallholder dairy cattle in Kenya. This was a cross-sectional study based on a single visit to farms to collect serum samples and other descriptive farm-level and animal-level information. Testing for BVDV Ag and Ab was conducted on serum samples from 320 dairy cows and heifers, with CSFV Ag and Ab testing conducted on a subset of 133 and 74 serum samples, respectively. CSFV testing was based on BVDV test results and the availability of enough sample volume from farms that kept pigs. The Ag and Ab tests utilized IDEXX ELISA for both BVDV and CSFV. For the 74 samples with Ab tests for both viruses, 40 (54.0%) were BVDV Ab positive, while 63 (85.1%) were CSFV Ab positive. Of the 40 BVDV Ab positive samples, 36 cattle (90.0%) tested positive for CSFV Ab. However, of the 34 BVDV Ab negative samples, 27 (79.4%) were CSFV Ab test-positive. For the 133 samples with Ag tests for both viruses, 125 (94.0%) were BVDV Ag positive, while 2 (1.5%) samples were CSFV Ag positive. None of the eight BVDV Ag negative samples was positive for CSFV Ag and only two (1.6%) of the 125 BVDV Ag positive samples were positive for CSFV Ag. The results indicate either substantial cross-reactivity of the two Ab ELISA tests, or reactivity with some other protein in the samples that led to the positive Ab test results. There was only limited evidence for cross-reactivity of the two Ag ELISA tests. We recommend that Pestivirus genus cross-reactivity be considered when interpreting BVDV ELISA results in cattle, more for Ab than Ag tests. Further research is needed to clarify the levels of cross-reactivity between BVDV and other Pestivirus Ag and Ab tests from animals on mixed-species farms. Keywords: antibody, antigen, bovine viral diarrhea virus, classical swine fever virus, cross-reactivity, smallholder dairy.

Chickens are considered as the main source of Salmonella, with infection potentially spreading to the public through outlets. The study aimed to investigate poultry shops for Salmonella enterica resistant to extended-spectrum cephalosporins-resistant (ESCR) and carbapenems-resistant (CR). Samples were collected from chicken giblets, water tanks, and workers at retail shops. Salmonella was isolated and serotyped; the presence of invA, stn, ompA, and ompF was determined using polymerase chain reaction (PCR). The isolates were tested for ESCR and CR by a disk-diffusion test; a confirmatory extended-spectrum β-lactamase (ESBL) test was performed by combinational disk-diffusion test with clavulanic acid. The resistant isolates were screened for ESBL (blaTEM, blaSHV, blaCTX-M, and blaOXA-1), AmpC blaCMY-2, and carbapenemase (blaKPC, blaNDM, and blaOXA-48) genes using PCR. S. enterica was isolated from chicken giblets (13/129) and the 13 isolates were ESCR. Based on the confirmatory ESBL test and CR, the 13 isolates were classified into the following resistance phenotypes: ESBL-producing and CR (n=4), ESBL-producing (n=1), non-ESBL-producing and CR (n=6), and non-ESBL-producing (n=2). All the five isolates with ESBL-producing phenotype carried predominantly blaTEM, blaSHV, and blaCMY-2. Regardless of being phenotypically CR, none of these isolates carried any of the tested carbapenemase genes. Surprisingly, the isolates with non-ESBL phenotype were found to carry blaTEM, blaSHV, and blaCMY-2. The blaKPC was present mainly in the isolates with non-ESBL and CR phenotypes. Interestingly, two isolates of the non-ESBL and CR phenotype showed resistance to cefepime, the fourth generation cephalosporins. Salmonella was also recovered from the water tanks (2/7) and the workers (2/16). The four isolates were ESCR and showed a non-ESBL-producing and CR phenotype; they harbored blaTEM, blaSHV, blaOXA-1, and blaKPC. The blaCMY-2 was found in one isolate from water and one from humans. All Salmonella isolates carried invA, stn, ompA, and ompF. Virulent ESCR S. enterica were identified in retail shops. The isolates carried blaCMY-2 and ESBL-genes, with a high proportion showing CR. Transmission of such strains to humans through food leads us to recommend regular inspection of retail outlets for antibiotic-resistant bacteria. Keywords: carbapenems, extended-spectrum β-lactamase, extended-spectrum cephalosporin-resistant, plasmid AmpC, Salmonella enterica.

Angiostrongylus eosinophilic meningitis is caused by larvae of the rat lungworm Angiostrongylus cantonensis. It manifests as meningitis, radiculitis, cranial nerve abnormalities, and encephalitis, which can be fatal. A flavan-3-ol compound isolated from the bark of Calophyllum macrophyllum Scheff. has several medicinal properties, including antioxidant, anti-inflammatory, antidiabetic, and antibacterial activities. This compound is stronger than other types of flavan-3-ols such as catechin. This study aimed to identify the hydroxylation pattern of this flavan-3-ol compound and evaluated its potential as an anti-meningitis drug, using an in silico approach through pharmacophore and molecular docking methods. Pharmacokinetic and toxicological data were analyzed and supported by the server http://www. swissadme.ch/index.php and https://tox-new.charite.de/protox_II/index.php. The hydroxylation pattern of the flavan-3-ol compound was identified using shear reagents (MeOH, NaOH, NaOAc, HCl, and AlCl3). The CviR receptor (pdb id.3QP5) was used in the in silico approach, and seven ligands were downloaded from PubChem in "SMILES" format. The spectroscopic analysis conducted using the shear reagents confirmed that the flavan-3-ol compound has a "p-diOH" pattern on the cinnamoyl ring. Pharmacophore analysis revealed this compound "hit" with pharmacophore features, and molecular docking analysis showed that this compound has a strong affinity with both receptors. The flavan-3-ol compound is a potential drug candidate for meningitis caused by pathogenic bacteria and the worm A. cantonensis. This result was supported by the pharmacokinetic profile, which had a very low toxicity level to the host. However, further investigation is required to confirm the data in vitro and in vivo. Keywords: Angiostrongylus cantonensis, Flavan-3-ol, molecular docking, pharmacophore.

Generally, rectal body temperature (BTrectum) is used to prefer as core body temperature in dogs. However, this procedure is time- and labor-consuming with stress induction. Therefore, infrared auricular temperature (BTear) and surface temperature (ST) could be applied to estimate BTrectum. This study aimed to estimate BTrectum from BTear or ST in various areas and determined the factors that influenced the accuracy of prediction equations. Under controlled temperature (n=197) and ambient temperature (n=183), the parameters BTrectum, BTear, and ST at internal pinna, auricular canal, lateral aspect of shoulder, hip, axillary area, inguinal area, footpad, and anal area (STrectum) were measured. In addition, temperature and humidity levels of the surrounding environment were recorded. The correlation between each measurement technique was calculated. The BTrectum prediction equation was created using all measured data and several influencing factors (environmental condition, size, coat type, and body condition score [BCS]). The highest correlation with BTrectum was observed for BTear (r=0.61, p<0.01), which was similar to STrectum (r=0.61, p<0.01). Based on multiple linear regression model results using BTrectum as the dependent variable, BTear or STrectum were first selected as independent variables in all estimation equations. Ambient temperatures (R2=0.397), small breed (R2=0.582), long hair (R2=0.418), and/or a BCS of 2 (R2=0.557) provided the highest coefficients of determination of the prediction equation. The most appropriate predictors for estimating BTrectum were STrectum and BTear, which were impacted by the dog's signalments and the environment. To obtain satisfactory outcomes, the equation must be selected depending on the dog's signalments and the environmental conditions. However, based on the findings of this investigation, the accuracy remains low in several equations, and further studies are needed to improve the accuracy of the equation, mainly by increasing the sample size and developing a specific equation for each dog's signaling and environmental condition. Keywords: auricular temperature, body surface temperature, dog, infrared surface temperature, rectal surface temperature.

Hemotropic Mycoplasmas are small epierythrocytic bacteria that cause infectious anemia in several livestock species and in humans. Several reports have been made on hemoplasma infections in the south and north of Iraq, but there have been no studies in the middle Euphrates of Iraq. This study aimed to evaluate the presence of hemoplasma species in cattle in Al-Qadisiyah Province, Iraq. Two hundred blood samples were collected from cattle with pale mucous membrane from regions with heavy tick endemicity. The samples were analyzed for the presence of Rickettsia pathogens using thin blood smears and the Diff-Quik stains. All the samples were also examined using polymerase chain reaction (PCR) to amplify the 16S ribosomal ribonucleic acid (rRNA) gene to confirm the presence of the smear-identified microorganisms. Ten PCR positive samples were subjected to 16S rRNA partial gene sequencing to identify the species. The findings uncovered positivity in 68 (34%) blood smears. PCR revealed positive confirmation in 18 (9%) of the 200 blood samples. Mycoplasma wenyonii and Candidatus mycoplasma hemobos were identified from 10 PCR positive samples. The nucleotide sequences of the isolates were closely related to isolates from cattle, buffalo, and dogs in Vietnam, Cuba, India, and Germany. Bovine hemoplasma infections are prevalent in cattle in the Al-Qadisiyah Province in Iraq. Our results may have significance for the development of control programs. Keywords: hemoplasma spp., microscopic, molecular diagnosis, Mycoplasma wenyonii, Iraq.

The world and the way things are done have changed, from selling clothing in brick-and-mortar stores to online shopping through social media platforms. Population growth has significantly contributed to an increased clothing demand, which, in turn, has increased the demand for animal skin. Traditional markets, also known as wet markets, are considered as major zoonotic disease reservoirs due to human and animal contact. Some groups and individuals continue to believe in traditional medicine and clothing that is made from animal skin, and such beliefs are more accessible with the presence of wet markets. Hence, animal poaching and trafficking have increased to meet the high demands, primarily in the Western world. Poverty is a well-known motivation to commit a crime. Conservationists should not only look at the animal regulation site to propose a solution to animal poaching and trafficking but should also consider communal poverty. Thus, this review aimed to highlight the role of wet market and animal skin fashion on animal welfare and human health.

Specific tumor biomarkers are useful for the early diagnosis of cancer or can predict the recurrence of neoplastic disease in humans and animals. Lymphoma in dogs could be classified into B-, T-, and NK-cell origins. T-cell lymphoma has the worst prognosis with a shorter survival time and disease-free interval. This study aimed to identify the differential serum protein expressions of canine B- and T-cell lymphomas compared with healthy dogs using a tandem mass tag (TMT)-based quantitative proteomics. Serum samples were collected from 20 untreated canine lymphomas (14 B-cells and 6 T-cells) and four healthy control dogs. Sera peptides from each sample were processed for TMT 10-plex tagging and analyzed using liquid chromatography-mass spectrometry (MS). Differential proteome profiling was then compared between lymphoma and control. We discovered 20 elevated and 14 decreased serum proteins in the lymphoma group relative to the healthy group. Six candidate increased proteins in canine lymphomas were beta-actin cytoplasmic 1 (ACTB, p=0.04), haptoglobin (p=0.002), beta-2 microglobulin (β2M, p=0.007), beta-2 glycoprotein 1 (APOH, p=0.03), metalloproteinase inhibitor 1 (TIMP-1, p=0.03), and CD44 antigen (p=0.02). When compared between B- and T-cell lymphomas, B-cell phenotypes had upregulated immunoglobulin (Ig) heavy chain V region GOM (p=0.02), clusterin (p=0.01), apolipoprotein C1 (APOC1, p=0.05), and plasminogen (p=0.02). These findings were investigated quantitative serum proteomes between B- and T-cell lymphomas using TMT-based MS. ACTB, β2M, APOH, TIMP-1, CD44 antigen, Ig heavy chain V region GOM, and APOC1 are novel candidate proteins and might serve as a lymphoma biomarker in dogs. However, evaluation with an increased sample size is needed to confirm their diagnostic and prognostic ability. Keywords: B-cell lymphoma, dog, tandem mass tag proteomics, T-cell lymphoma.

Japanese encephalitis (JE) is a zoonotic infectious inflammatory brain disease caused by the JE virus (JEV). Considerable research into the seroprevalence of JE in domestic animals has been conducted, but there have been no reports of its occurrence in wild animals, including long-tailed macaques (Macaca fascicularis). This study aimed to estimate the seroprevalence of JEV infection and its determinants in long-tailed macaques in Bali and the prevalence of mosquito vectors. Blood samples (3 mL) were collected from a population of M. fascicularis (92 heads) inhabiting a small forest with irrigated rice field nearby (wetland area) in Ubud, Gianyar, and from two populations in dryland areas with no wet rice field (Uluwatu, Badung, and Nusa Penida, Bali Province, Indonesia). The collected sera were tested for antibodies against JEV using a commercially available enzyme-linked immunosorbent assay kit (qualitative monkey JE Immunoglobulin G antibody kit). The seropositivity of the antibodies was then compared based on different variables, namely, habitat type, age, and sex. The seroprevalence of the JEV antibodies in all the samples tested was found to be 41.3%. The seropositivity of the monkey serum samples collected from the wetland area was 46.4%, which was higher than the seropositivity of the sera samples collected from the dried field areas (1.25%). Monkey sera collected from the wetland areas were 6.1 times (odds ratio [OR]: 6.1; 95% confidence interval [CI]: 0.71-51.5, p>0.05) more likely to be seropositive compared to the monkey sera collected from the dried field areas. Meanwhile, female monkeys were 1.79 times (OR: 1.79; 95% CI: 0.76-4.21; p>0.05) more likely to be seropositive to JEV than males. Similarly, juvenile monkeys were 2.38 times (OR: 2.38; 95% CI: 0.98-5.79); p>0.05) more likely to be seropositive against the JEV than adult monkeys. However, none of these differences achieved statistical significance. Regarding the JEV mosquito vector collection, more Culex mosquitoes were found in the samples from the wetland areas than from the dried field areas. The study confirms the existence of JEV infection in long-tailed macaques in Bali. There were patterned seropositivity differences based on habitat, age, and sex of the monkeys, but these were not significant. The possibility of monkeys as a JEV reservoir and the presence of the mosquitoes as the JEV vector are suggested but require more study to confirm. Keywords: enzyme-linked immunosorbent assay, Japanese encephalitis virus, Macaca fascicularis, seroprevalence.

Bone marrow-derived mesenchymal stem cells (BM-MSCs) transplantation and their hepatogenic differentiated cells (HDCs) can be applied for liver injury repair by tissue grafting. Regenerative potentiality in liver cirrhosis models was widely investigated; however, immunomodulation and anti-inflammation in acute hepatitis remain unexplored. This study aimed to explore the immunomodulatory and evaluate twice intravenous (IV) or intrahepatic (IH) administration of either BM-MSCs or middle-stage HDCs on aflatoxin (AF) acute hepatitis rat model. BM-MSCs viability, phenotypes, and proliferation were evaluated. Hepatogenic differentiation, albumin, and a-fetoprotein gene expression were assessed. AF acute hepatitis was induced in rats using AFB1 supplementation. The transplantation of BM-MSCs or their HDCs was done either by IV or IH route. Hepatic ultrasound was performed after 3-weeks of therapy. Cytokines profile (tumor necrosis factor-α [TNF-α], interleukin [IL]-4, and IL-10) was assessed. Hepatic bio-indices, serum, and hepatic antioxidant activity were evaluated, besides examining liver histological sections. Acute AFB1 showed a significant increase in TNF-α (p<0.01), liver enzyme activities (p<0.05), as well as decrease in IL-4, IL-10, and antioxidant enzyme activities (p<0.05). Cytokines profile was ameliorated in groups treated with IV and IH BM-MCs, showed a negative correlation between IL-4 and TNF-α (p<0.05), and a positive correlation between IL-10 upregulation and TNF-α (p<0.01). In IV HDCs treated group, positive correlations between IL-4 and IL-10 downregulation and TNF-α were observed. However, in IH HDCs group, a significant positive correlation between IL-4 and IL-10 upregulation and TNF-α, were recorded (p<0.05). In addition, IV BM-MSCs and IH HDCs treatments significantly increased antioxidant enzymes activity (p<0.05). IV and IH BM-MSCs significantly ameliorated liver transaminase levels, whereas IH HDCs significantly ameliorated alanine aminotransferase activity and nitric oxide concentration (p<0.05). The administration routes of BM-MSCs did not demonstrate any significant difference; however, the IH route of HDCs showed significant amelioration from the IV route. On the other hand, it showed noticeable anti-inflammatory and immunomodulatory improvements in aflatoxicosis rats. Therefore, it can be concluded that acute hepatitis can be treated by a noninvasive IV route without the expense of hepatogenic differentiation. Further research using clinical trials that address several problems regarding engraftment and potentiation are needed to determine the optimal manipulation strategy as well as to achieve better long-term effects. Keywords: acute hepatitis, bone marrow mesenchymal stem cell, cytokines, hepatogenic differentiation, rat model.

Micronutrients such as essential amino acids in chicken feed must be balanced to promote optimal development. The balance of the amino acids arginine and lysine in chicken feed is particularly important. This study aimed to examine the effect of the ratio of L-arginine to L-Lysine HCl on growth performance and ileum morphology of native chickens aged 2-14 weeks-old. One hundred and eighty 2-week-old native chickens which initial weight 78.10±4.97 g were classified into six treatments and five repetitions using a completely randomized design. Treatments were based on the ratio of arginine to lysine in the feed: T1 (0.50% L-arginine: 0.85% L-lysine HCl); T2 (0.75% L-arginine: 0.85% L-lysine HCl); T3 (1.00% L-arginine: 0.85% L-lysine HCl); T4 (0.50% L-arginine: 1.00% L-lysine HCl); T5 (0.75% L-arginine: 1.00% L-lysine HCl); and T6 (1.00% L-arginine: 1.00% L-lysine HCl). Groups T3 and T6 had the highest feed consumption (42.06±0.29 and 42.78±0.72 g/bird/day, respectively), while Group T6 had the highest body weight and body weight gain rate (1505.60±103.20 kg/bird and 16.99±1.24 g/bird/day, respectively). Groups T3 and T6 also had the highest carcass weight (916.16±46.99 and 947.18±62.32 g/bird, respectively). The best feed conversion was seen for Groups T3, T5, and T6 (2.55±0.14, 2.50±0.20, and 2.53±0.19, respectively). For ileum morphometry, the highest villus height occurred in Groups T2, T3, T5, and T6 (962.80±23.31, 982.80±10.03, 972.80±18.99, and 989.80±10.69 μm, respectively); and Group T6 had the highest crypt depth and villus width (340.80±11.52 and 302.00±4.00 μm, respectively). Statistical analysis indicated significant differences among the treatment groups for all variables examined (p<0.05). The highest ratio of arginine-lysine was associated with the largest increase in native chicken feed consumption, body weight gain, feed conversion, and carcass weight, as well as villus height and width, and crypt depth in the ileum. Overall, an arginine-lysine ratio of 0.8-1.20 promoted optimal growth of native chickens aged 2-14 weeks. In the future, it is important to increase the arginine-lysine ratio with low feed protein levels in native chickens. Keywords: growth performance, ileum morphology, L-arginine, L-lysine HCl, native chickens.

Pullorum is an acute and chronic disease caused by Salmonella pullorum, often infecting chicken farms. Pullorum disease treatment using antibiotics that do not follow the control dose can cause bacteria to become antibiotic-resistant. Meniran contributes to inhibiting and antagonizing bacteria and can increase the efficiency of chicken feed because of its bioactive compounds, including alkaloids, flavonoids, tannins, and saponins. This study aimed to determine the activity of Meniran extract (Phyllanthus niruri Linn.) in broilers infected with S. pullorum. In vitro study that was conducted includes phytochemical test, diffusion, and dilution methods using Meniran extract at 5%, 10%, 20%, and 40% concentrations and tylosin at 2% concentration. The data of the dilution method (minimum inhibitory concentration [MIC] and minimum bactericidal concentration [MBC]) were processed using probit analysis to determine LC50. In vivo study was conducted by randomly dividing 20 broilers into five treatment groups, four per group. The chickens (except in group P0–) were infected with S. pullorum aged 14 days. Then, the treatment was conducted according to the divided groups when the chickens were aged 21-34 days. The said treatments are P0– (uninfected S. pullorum and unadministered with Meniran extract), P0+ (infected with S. pullorum and unadministered with Meniran extract), and P1, P2, and P3 (infected with S. pullorum and administered with Meniran extract with 5%, 10%, and 20% concentrations, respectively). Data from the phytochemical test were analyzed as descriptive. The data from the diffusion method were analyzed using one-way analysis of variance (ANOVA) and Duncan's test. Then, the results of broilers' performance were analyzed using ANOVA and Duncan's test. The phytochemical test showed positive for alkaloid, tannin, saponin, flavonoid, and steroid/triterpenoid. The diffusion method formed the largest zone at 40% concentration with 15.6 mm, while 20%, 10%, and 5% had average of 13.15 mm, 8.38 mm, and 5.8 mm, respectively. The dilution method (MIC and MBC) exhibited the antibacterial ability of Meniran extract against S. pullorum at 20% dose and LC50 14.118% concentration. The Meniran extract administration in broilers exhibited improved performance of chickens infected with S. pullorum, with the administration of 20% dose of Meniran extract showing the best result. About 20% concentration Meniran extract can serve as an antibacterial agent and showed the best results in broilers infected with S. pullorum. Keywords: broiler chicken performance, food security, Phyllanthus niruri Linn., Salmonella pullorum.

Rabies is considered a highly fatal zoonotic disease and many deaths in humans have been associated with dog bites. This study was designed to prepare an oral anti-rabies vaccine in the form of baits to eliminate the disease in free-roaming dogs and subsequently protect humans from dog bites. The Evelyn Rokintniki Abelseth (ERA) rabies virus strain was propagated in baby hamster kidney cell cultures and adjusted to the recommended dose for application. Four forms of oral baits were employed with the rabies vaccine, which was evaluated for safety, acceptability, and potency in different dog groups. Enzyme-Linked Immunosorbent Assay (ELISA) and the serum neutralization test (SNT) were used to determine the protective rabies antibody titer in the sera of vaccinated dogs. According to the results, a dose of 3 mL of the ERA strain, containing a viral titer of 107.6 TCID50/mL, induced a mean antibody titer of 25.6 by SNT, and the PI% was 75.7 by Block ELISA, providing a protective level of the rabies antibody in 100% of vaccinated dogs. All used baits were found to be safe, inducing no abnormal general post-vaccination signs (the signs are limited to mild fever, mild loss of appetite, and mild-to-moderate loss of energy for 24-36 h after vaccination). It was found that most of the accepted and highly potent bait types consisted of a mixture of wheat flour, vegetable oil, sodium alginate, corn starch, meat meal, cellulose gum, and water. This dog meal was covered with bran and edible wax to seal the bait cavity after inserting the vaccine sachet. This bait was able to induce a protective level of rabies antibodies in 100% of vaccinated dogs after receiving one bait/dog. Hence, such a bait could be recommended for use in the protection of free-roaming dogs and the elimination of the disease. Keywords: enzyme-linked immunosorbent assay, oral baits, potency, rabies, serum neutralization test.

Bovine tuberculosis (TB) is a zoonotic disease that causes huge economic losses. This study aimed to compare the result obtained from the single intradermal test, conventional methods (culture and microscopy), gamma-interferon (IFN-γ) assay, and indirect enzyme-linked immunosorbent assay (ELISA) to diagnose bovine TB. This study evaluated 2913 animals from milk farms in Cairo, El-Sharkia, and El-Qalyubia Governorates by single intradermal cervical tuberculin technique (SICTT), ELISA, and IFN-γ assay. Of the 2913 dairy cows surveyed, 3.7% yielded positive results. Culture prepared samples on Lowenstein-Jensen and Middlebrook 7H10 agar media yielded 52 (1.85%) isolates of Mycobacterium spp. from 2805 milk samples that yielded negative tuberculin reactions and 56 (51.85%) isolates of Mycobacterium spp. were recovered from 108 lymph node samples from positive cases. ELISA analysis of the sera of 108 positive SICTT reactors revealed that 94 (87.03%) and 97 (89.81%) animals were positive for bovine purified protein derivative (PPD-B) antigen and commercial polypeptide antigen, respectively. IFN-γ assays were performed on whole blood samples collected from positive SICTT reactors and showed that 103 (95.37%) animals were positive. M. tuberculosis complex may be isolated from raw milk and not all infected animals shed mycobacterial bacilli in their milk. The use of polypeptide antigen in ELISA provides better diagnostic efficacy than PPD-B antigen. The IFN-γ assay is more sensitive than both SICTT and ELISA. It should be used in parallel with SICTT to allow the detection of more positive animals before they become a source of infection to other animals and humans. Keywords: bovine tuberculosis, enzyme-linked immunosorbent assay, Gamma-interferon assay, Mycobacterium tuberculosis, single intradermal cervical tuberculin skin test.