Objective: This study analyzes the mycobiome in wild and captive Sumatran orangutans. Materials and Methods: Nine orangutan feces samples from the wild and nine from captivity were divided into three repeats from 11- to 15-year-olds in good health. The Illumina platform for analysis of ITS bioinformatics was used according to the Qiime2 and CCMetagen approaches. Results: Wild Sumatran orangutans include 53% Ascomycota, 38% uncultured fungi, and 4% Basidiomycota. Orangutans in captivity are 57% Ascomycota, 26% uncultured fungi, and 2% Basidiomycota. Based on genus level, uncultured Neurospora (31%), Penicillium (10%), Aspergillus (3%), Fusarium (3%), Candida (2%), Cutaneotrichosporon (2%), and Limonomyces (2%) are found in wild orangutans. The most prevalent genus among captivity orangutans is Aspergillus (32%), followed by fungal sp. (11%), Lasiodiplodia (18%), Devriesia (2%), and Sordariomycetes (2%). According to the Chao1 diversity index and Shannon and Simpson, there was no significant difference between wild and captive Sumatran orangutans. Conclusion: Neurospora is unique to wild Sumatran orangutans, although Aspergillus predom¬inates in captive orangutans. We hypothesize that the gut mycobiome of wild orangutans will resemble that of orangutans in captivity. The excellent range of food sources in the forest does not result in the prevalence of fungi in the typical gut microbiome. [J Adv Vet Anim Res 2023; 10(2.000): 228-236]

Objectives: Ducks suffer a huge economic loss as a result of infections with Pasteurella multocida and Riemerella anatipestifer, which cause high morbidity and mortality. Because these patho¬gens induce similar clinical symptoms when coinfections occur, it is very difficult to differentiate between them based just on clinical signs. Hence, these major pathogens must be quickly and accurately detected. Materials and Methods: A total of 104 birds ranging from 2 days to 4 weeks old were collected from Egyptian farms, and the outcomes were compared statistically. Conventional cultural iden¬tification procedures and a direct multiplex polymerase chain reaction assay were utilized to recognize both pathogens in a single tube reaction simultaneously. Then, the obtained isolates were characterized phenotypically and genotypically. Results: Clinical signs appear at 2–4 weeks of age with respiratory distress (dyspnea), white fluid feces, and stunting. The scrutinized data demonstrated a significantly higher detection rate by PCR directly compared to classical culture procedures. Pasteurella multocida was detected only by PCR. The disc diffusion technique against ten antibiotics showed absolute susceptibilities to amik¬acin, doxycycline, and florfenicol. High levels of beta-lactam resistance were observed. Riemerella anatipestifer isolates were screened for pathogenicity and plasmid-borne blaTEM genes. All six isolates harbored five virulence genes: aspC, RA46, m28, pstS, and Nlp/P60. Moreover, blaTEM was identified into four isolates and deposited to GenBank with accession numbers OP347083, OP347084, OP347085, and OP347086. Conclusion: These results suggest advanced PCR assays can be applied to the field for rapid and valuable diagnosis of two significant pathogens and focus on the worth of ducks in the propaga¬tion of transferable antibiotic resistance genes into the environment. [J Adv Vet Anim Res 2023; 10(2.000): 211-221]

Objective: Microfilariae parasites are common in tropical regions, and some species are reported as potentially zoonotic. The diagnosis of filarial infection in dogs by cytology or hematologic tech¬niques showed lower sensibility and specificity, which may result in misdiagnosis. Thus, molecular techniques seem to be an alternative to identifying and detecting microfilariae infections. On the other hand, lymphoma is one of the main tumors in domestic animals, with a high prevalence in domestic canines. This study aims to report a mixed infection with microfilariae in a dog with lymphoma, emphasizing its diagnosis and the possible role of this infection in the development of the neoplasia. Materials and Methods: An 8-year-old male mixed breed dog was referred to consultation due to the presence of lethargy, recumbency, skin ulceration lesions, nonspecific pain manifestations, emesis, myoclonus in the left temporalis muscle, and seizures. Routine blood and biochemistry tests were normal, and cytology of the skin evidenced a microfilariae infection. The dog died due to a cardiorespiratory arrest, and tissue sampling was done for histopathology and molecular analysis at the necropsy examination. Results: Skin lesions were related to a microfilarial pyogranuloma related to Acanthocheilonema reconditum. Histopathology of the spleen and liver revealed a diffuse lymphoma composed of blast cells and large lymphocytes, distributed diffusely in the parenchyma and surrounding the vasculature. In the skin, microfilariae were seen in some superficial capillaries. Conclusion: This study describes a microfilariae mixed infection with A. reconditum and Dirofilaria immitis in a dog with a lymphoma and its molecular detection. To the knowledge of the authors, this is the first report of a mixed microfilariae infection in a tumor of a dog and highlights the use of molecular techniques, i.e., polymerase chain reaction, for an accurate diagnosis.

Objective: This study was conducted to evaluate the effect of composting on the count of Salmonella spp., Clostridium perfringens, and New Castle virus (NDV) isolated from broilers' litter. Moreover, to verify the impact of compost thermal stress on the expression of thermal genes harbored in the isolated bacteria. Materials and Methods: The prevalence of enteric aerobic and anaerobic infections by Salmonella spp., C. perfringens, and viral infections by NDV were investigated in litter samples collected from 100 broiler flocks by conventional methods and polymerase chain reaction. Results: The samples were positive for Salmonella spp., C. perfringens, and NDV, with prevalence rates of 60%, 55%, and 30%, respectively. An experiment to study the effect of compost on the microbiological quality of litter was applied using five compost heaps with an initial average count of Salmonella typhimurium (3.2 × 105CFU CFU/gm), C. perfringens (6.4 × 105 CFU/gm), and an average titer NDV (105.5 embryo infectious dose50/gm). The microbiological count of heaps after 15 days of composting revealed a reduction in the count of S. typhimurium and C. perfringens by 4 log10 CFU/gm and 3 log10 CFU/gm, respectively. Moreover, the hemagglutinating test revealed no detection of NDV after 15 days of composting. A high degree of downregulation of expression of the thermal genes, dnaK in S. typhimurium isolates and cpe gene in C. perfringens isolates, was detected by quantitative reverse transcription PCR. Conclusion: The reduction of pathogen counts, the simplicity, and the low cost associated with composting for only 15 days advocate the recommendation for raising awareness of composting as a routine biosecurity measure to prevent the spreading of infection and promote its safe use in agribusiness.

Objective: The study aimed to determine the potential anthelmintic activity of the ethyl acetate extract of Eleusine indica that will result in an effective reduction in fecal egg per gram (EPG) counts in naturally infected goats compared to the commercial anthelmintic levamisole. Materials and Methods: The experimental animals were 21 goats naturally infected with gas¬trointestinal nematodes. The goats were divided into groups that were given a single dose of E. indica extract. Five concentrations of E. indica were tested for anthelmintic activity: 100, 200, 300, 400, and 500 mg extract/kg body weight. Fecal sample collection was done before treatment, during the first treatment, and every week thereafter for 28 days post-treatment (dpt). A modi¬fied McMaster technique was used to determine the EPG of feces, and the mean efficacies of the extracts were compared with those of the commercial anthelmintic levamisole. Results: As early as 7 dpt, there was an observed reduction in the epg counts after the administration of E. indica extracts across all concentrations. Administering 500 mg of extract/kg body weight resulted in a maximum efficacy of 56.21%. However, the efficacy achieved was lower than that of levamisole (96.83%). Conclusion: The results show that the E. indica extract can reduce the fecal EPG counts of natu¬rally infected goats, thus creating a potential natural anthelmintic that can be developed further.

Objective: This research aimed to assess the prevalence of caprine pasteurellosis, isolate and identify pasteurellosis (Mannheimia haemolytica and Pasteurella multocida) in pneumonic goats, and discover the main bacterial cause of pneumonia. Materials and Methods: One hundred and five samples (94 nasal swabs and 11 lung tissues) from goats suspected of having pneumonia were taken and transferred aseptically to the laboratory. Following the processing of the collected samples, Pasteurella spp. was isolated with the aid of plate culture methods. Biochemical characteristics were used to identify all bacterial isolates, which were then verified by polymerase chain reaction (PCR). Antimicrobial susceptibility testing was also carried out to evaluate the sensitivity profiles of various antibiotics. The Pasteurella hae¬molytica serotype-specific antigen (PHSSA) gene was used to identify isolates of M. haemolytica, and the KMT1 gene was used to identify isolates of P. multocida. Results: From the 105 clinically suspicious samples, 51 (48.57%) were identified to be Pasteurella spp. through bacteriological testing and also by PCR targeting the 16S rRNA gene. Of these, 47.87% (45/94) were nasal swabs, and 54.55% (6/11) were lung tissues. Among confirmed sam¬ples, 70.59% (36/51) were identified as M. haemolytica, and 29.41% (15/51) were identified as P. multocida. Resistance to tetracycline, streptomycin, oxytetracycline, gentamicin, and ceftriax¬one was found in 50%–83% of the isolates. In addition, PCR identified the PHSSA and KMT1 genes from isolates of P. multocida and M. haemolytica, respectively. Conclusion: The present study revealed that M. haemolytica and P. multocida primarily caused pasteurellosis in pneumonic goats in Bangladesh. However, when treating these animals, the proper choice of antimicrobials should be made to control this disease.

Objective: This study aims to determine the protein profile based on molecular weight (MW) and testosterone levels in seminal plasma (SP) that correlates to the semen quality of candidate Madura bulls. Material and Methods: A total of 10 male candidate madura bulls underwent semen evaluation (motility, viability, membrane plasma integrity (MPI), and sperm concentration). The centrifuge was run at 1,200 rpm (4°C) for 20 min to collect SP. SP testosterone levels were measured using an Enzyme-linked immunosorbent assay. The characterization of SP proteins in Madura bulls was done using 1D sodium dodecyl-sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis. All parameters were analyzed using Pearson correlation analysis. Result: The results of the SDS-PAGE analysis found eight protein bands with the highest MW of 110 kDa and the lowest of 12 kDa. The mean and SD of SP testosterone levels were 20.58 ± 8.56 ng/ml, motility 59.32% ± 20.14%, viability 67.45% ± 20.22%, MPI 32.77% ± 16.52%, and sperm concentration 1,002.64 ± 429.33 106/mm3. Proteins with MWs of 110 and 91 kDa significantly correlated with MPI, and 110 kDa negatively correlated with sperm concentration (p < 0.05). Proteins with MWs of 73 and 36 kDa significantly correlated with SP testosterone levels, while proteins with MWs of 29 kDa significantly correlated with sperm viability (p < 0.05). Conclusion: The expressed protein fraction based on MW is closely related to the quality of semen, so it has the potential to be a biomarker of semen quality. Further research is needed to determine the specific proteins in certain fractions.

Objective: This research aims to identify the effect of various organic solvents such as n-hexane, ethyl acetate (EtOAc), and methanol (MeOH) on the antioxidant and antibacterial activities of Curcuma zedoaria extract, against three Gram-positive bacteria, namely Staphylococcus aureus, Bacillus subtilis, and Streptococcus pneumoniae, and three Gram-negative bacteria, namely Escherichia coli, Salmonella typhi, and Pseudomonas aeruginosa. Materials and Methods: As much as 1 kg of white turmeric rhizome (C. zedoaria) was extracted two times for 24 h using 3 l of MeOH before evaporating. The extract was then fractionated using n-hexane six times per 2 h, with each volume of 500 ml, and continued with the EtOAc fractionation. The MeOH fraction was added to 300 ml of water before adding 400 ml of EtOAc. Once the fractionation process was complete, all fractions were concentrated using a rotary evaporator. Results: The C. zedoria extract fractioned using MeOH produces alkaloids, phenolics, flavonoids, saponins, and coumarin compounds. The fractionation with EtOAc also produces alkaloids, phenolics, flavonoids, saponins, coumarin compounds, and triterpenoids. Meanwhile, fractionation with n-hexane only produces alkaloids and triterpenoid compounds. EtOAc and MeOH fractions had good activity in reducing free radicals produced by 2,2-diphenyl-1-picrylhydrazyl (DPPH), with an average IC50 value of 153.49 ± 2.66 and 185.77 ± 3.91 ppm, respectively. In contrast, the n-hexane fraction has weak antioxidant activity with an IC50 value of 837.92 ± 5.32 ppm. The n-hexane fraction has better activity compared to MeOH and EtOAc. The lowest concentration required was 2,500 ppm for all types of bacteria. Conclusion: Curcuma zedoaria extract produces alkaloids, phenolics, flavonoids, saponins, coumarins, and triterpenoids when fractionated with MeOH or EtOAc. Only alkaloids and triterpenoids are produced using n-hexane. EtOAc and MeOH fractions have good activity in reducing free radicals generated by DPPH, with an average IC50 value of 153.49 ± 2.66 and 185.77 ± 3.91 ppm, respectively. However, n-hexane has weak antioxidant activity, with an average IC50 value of 837.92 ± 5.32 ppm. All fractions have moderate antibacterial activity, but the extract of n-hexane from C. zedoary has better antibacterial activity compared to MeOH and EtOAc. The lowest concentration required is 2,500 ppm for all types of bacteria.

Objective: The goal of this study was to look at quinolone-resistant (QR) Escherichia coli (E. coli) from retail beef and poultry meat in Egypt by looking at the QR mechanisms in the resistant strains. Materials and Methods: In total, 120 samples of raw poultry meat (n = 60) and beef meat (n = 60) were purchased from Mansoura retail stores between January and March 2021, and evaluated microbiologically for E. coli. Then, an antimicrobial sensitivity test was applied to all isolates. The prevalence of QR E. coli with concern for the QR determinants, including quinolone resistance-determining regions (QRDRs) mutations, the plasmid-mediated quinolone resistance gene (PMQR), and the efflux pump activity were determined. Results: The total prevalence of E. coli was 34.2% (41/120). Noticeably, the prevalence of E. coli in poultry meat (40%, 24/60) was higher than that of beef (28%, 17/60). All strains were assessed for their antimicrobial susceptibility using the disc diffusion technique; the highest rate of resistance (100%) was displayed to clindamycin and cefuroxime, followed by ampicillin (97.6%), doxycycline (92.7%), amoxicillin-clavulanate (92.7%), nalidixic acid (NA) (80.5%), sulfamethoxazole/trimethoprim (70.7%), chloramphenicol (63.4%), gentamicin, and azithromycin (58.5% each). Multiple antimicrobial resistance (strains resistant to three or more antimicrobial classes) was displayed by 97.6% of E. coli isolates. Regarding QR, 37 isolates could resist at least one of the examined quinolones. Regarding PMQR genes, qnrS was determined in 70% (7/10) of QR E. coli, while qnrA, qnrB, and qnrD were not identified. While the mutations determined regions of QR in the resistant E. coli isolates, S83L was the most prevalent in gyrase subunit A either alone or combined with D87N and D87Y, and three isolates of QR E. coli isolates revealed a topoisomerase IV subunit mutation harboring S80I. 20% of the isolates displayed efflux activity, as NA showed a considerable difference between its zones of inhibition. Conclusion: The high prevalence of antimicrobial-resistant E. coli, with concern for QR strains harboring different resistance mechanisms in poultry meat and beef, threatens the public's health. Thus, standard manufacturing procedures and adequate hygiene conditions must be followed in all phases of meat preparation, production, and consumption, and public knowledge should be improved.

Objective: This study examines the effect of fermented milk containing the probiotic Pediococcus acidilactici BK01 on cholesterol and intestinal microbiota. Materials and Methods: 24 male rats weighing an average of 200 gm each spent 1 week in a cage adapting to their new environment. They were fed standard feed daily and were allowed to drink ad libitum. For 3 weeks, rats were divided into four groups (doses of fermented milk): M+ (control), M1 (0.35 ml), M2 (0.70 ml), and M3 (1.05 ml). The analysis includes bodyweight deter¬mination, serum biochemical analysis, and intestine microbiota analysis. Results: The results indicated that while P. acidilactici BK01 fermented milk did not affect body weight or high-density lipoprotein, it did have a beneficial effect on total serum cholesterol and triglyceride levels. Additionally, treatment of fermented milk with P. acidilactici BK01 has been shown to increase the total lactic acid bacteria (LAB) in the intestine, as indicated by changes in the intestinal villi. Conclusion: Administering fermented milk (P. acidilactici BK01, 1.05 ml) can reduce total serum cholesterol and increase the number of LAB in intestinal villi in experimental animals, so it has the potential to be a probiotic. [J Adv Vet Anim Res 2023; 10(1.000): 64-71]