Volume 16 | May
Research Article | 07 May 2023
Phenotypic diagnosis and genotypic identification of Bacillus cereus causing subclinical mastitis in cows
Rasha H. Eid, Noha E. Aref, and Eman S. Ibrahim

Volume-16 | Issue-5 | Article-1 | https://doi.org/10.14202/vetworld.2023.888-894

Preview Abstract
Bovine mastitis is a disease that affects dairy cows and impacts the global dairy industry. Bacillus spp. can infect the mammary gland during lactation, intramammary treatment, or dry cow therapy. This study aimed to isolate and identify Bacillus spp. in raw milk samples from cows with subclinical mastitis from dairy farms in Beheira, Giza, Alexandria, and Menoufia Governorate, Egypt. We also investigated their antibiotic sensitivity and detected the enterotoxigenic and antibiotic resistance genes. A total of 262 milk samples (15-20 ml each) were examined microscopically, biochemically, and phenotypically. A polymerase chain reaction was used for genotypic identification and detecting antibiotic-resistance and enterotoxigenic genes. Antibiotic sensitivity was tested using the agar well diffusion test. Bacillus cereus was identified in 47.7% of samples. Nhe and hblD enterotoxin genes were found in 93.64% (103/110) and 91.82% (101/110) of the samples, respectively. Tetracycline and β-lactam antibiotic-resistance genes were present in 0% (0/110) and 98.18% (108/110), respectively, of the samples. All isolates were resistant to cefepime, cefixime, and oxacillin, while they were susceptible to amoxicillin-clavulanic, chloramphenicol, ampicillin/sulbactam, and levofloxacin. These results highlight the need to promote awareness regarding B. cereus, the most common pathogen causing mastitis in Egyptian dairy cows. We also emphasized that antibiotic misuse during mastitis is a potential public health threat. Keywords: antibiotics, dairy cows, Egypt, lactation.
Research Article | 07 May 2023
Quantitative risk assessment for human Taenia solium taeniasis/cysticercosis exposure through consumption of pork in Mpwapwa District of Dodoma Region, Tanzania
Misheck A. Mulilo, Kabemba E. Mwape, Ethel M’kandawire, and Ernatus M. Mkupasi

Volume-16 | Issue-5 | Article-2 | https://doi.org/10.14202/vetworld.2023.895-911

Preview Abstract
Pig farming is a livelihood activity undertaken by many rural communities in Tanzania. However, pigs in rural communities become infected with Taenia solium, a zoonotic parasite leading to porcine cysticercosis (PCC). Thus, routine meat inspection is fundamental in ensuring that the meat consumed is PCC-free. However, routine meat inspection is constrained by low sensitivity as a diagnostic test. Regardless of its low sensitivity, at the local level, no substitute tool would potentially lead to meat being risk-free for human infection. This study aimed at quantifying the risk of humans getting exposed to T. solium taeniasis through the consumption of pork approved safe for human consumption by employing a quantitative risk assessment (QRA) model. A cross-sectional study employing a quantitative risk assessment technique was conducted to quantify the risk of human infection in Mpwapwa District through exposure to T. solium infection through consumption of officially inspected pork. The input parameters in this study were simulated in @risk software to obtain the risk of exposure and the risk factors for exposure to T. solium taeniasis. The risk of one getting exposed to T. solium taeniasis/cysticercosis (TSTC) through consumption of pork approved for human consumption was found to be 0.018 (95% confidence interval [CI] = 0.00–0.0250). Likewise, the probability that a cyst is localized in a pork portion was found to be the most influencing input risk factor of getting exposed to TSTC. Furthermore, the probability of developing T. solium taeniasis was estimated to be 0.73605 (95% CI = 0–0.950) when infected undercooked pork portion is consumed and 0.99652 (95% CI = 0.98161–0.99908) from consuming raw pork portion. Likewise, about 47 (95% CI = 42–52) people who consumed undercooked pork and 26 (95% CI = 22–30) who consumed raw pork would get infected in Mpwapwa District per year. The results from this study are anticipated to create public awareness of the problem and promote the use of one-health approach in the control and prevention of the consumption of infected pork. Keywords: cysticercosis, Mpwapwa, pork meal, quantitative risk assessment, Taenia solium, taeniasis.
Research Article | 07 May 2023
Phenotype and genotype of Boerka goats raised in Bali
I Nyoman Suyasa, I Wayan Suardana, I Gusti Agung Arta Putra, and Ni Nyoman Suryani

Volume-16 | Issue-5 | Article-3 | https://doi.org/10.14202/vetworld.2023.912-917

Preview Abstract
Boerka goats are the new breed in Bali targeted at meeting the increasing demand for lamb. They are meat-type goats created by crossing male Boer and female Kacang breeds. This new breed is expected to have good adaptation in a poor environment, such as Kacang goats and produce good quality lamb as Boer goats. Therefore, this study aimed to examine phenotypic and genotypic characteristics of Boerka goats raised in Bali. A total of 16 female Boerka goats at 2 years old collected from a group of livestock farmers in Sanda Village were used as samples. This observational study began with observations of qualitative characteristics and morphometric measurements of goats, followed by polymerase chain reaction (PCR) of the growth hormone (GH) gene using GH5F and GH5R primers. Polymerase chain reaction products were then sequenced and analyzed with the MEGA 11 program. The results showed that all Boerka goats had stuck-down ears, the heads were predominantly brown in color (62.5%), the body color pattern tends to be white (87.50%), and the tail color pattern was also dominated by white (62.5%). In morphometric terms, the samples were close to Kacang goats based on the body weight, head length, head width, chest width, depth, and circumference, left front leg circumference, ear length, ear width, tail length, tail width, and flank height except for head height, body length, horn length, and shoulder height. Analysis of the GH gene showed that Boerka goats had a nucleotide composition dominated by the purine base guanine (26.5%) and pyrimidine cytosine (31.8%). Furthermore, they formed a separate cluster with a genetic distance of 14.1% to the Anhui White breed from China, the Tibetan breed, and the Swiss Saanen breed. Boerka goats raised in Bali have phenotypes, including qualitative characteristics and morphometric measurements close to Kacang goats. As meat-type goats, they also form a separate cluster distinct from similar types worldwide. Keywords: Boerka goats, genotype, phenotype, raised in Bali.
Research Article | 07 May 2023
Effects of embryonic thermal manipulation on the immune response to post-hatch Escherichia coli challenge in broiler chicken
Mohammad Borhan Al-Zghoul, Ziad Waheed Jaradat, Mustafa M. Ababneh, Mohammad Ziad Okour, Khaled Musa Mohammad Saleh, Ayesha Alkofahi, and Mohammad Hussien Alboom

Volume-16 | Issue-5 | Article-4 | https://doi.org/10.14202/vetworld.2023.918-928

Preview Abstract
Thermal manipulation (TM), exposure to mild heat shock during embryogenesis, which is a critical developmental period of broiler chickens, improves tissue stability, oxidative stress response, and immune response during heat stress. Thermal manipulation could be more cost-effective than other methods to boost the immune response. This study aimed to evaluate the impact of TM during embryogenesis, concomitant with an Escherichia coli challenge, on body weight (BW), body temperature (Tb), and splenic mRNA expression of cytokines (Interleukin [IL]-1β, IL-2, IL-6, IL-8, IL-12, IL-15, IL-16, IL-18, and interferon [IFN]-γ) in poultry. A total of 740 fertile eggs were procured from a certified Ross broiler breeder. The eggs were divided into two incubation groups: the control and TM groups. The eggs in the control group were kept at 37.8°C air temperature and 56% relative humidity (RH) during incubation; eggs of the TM group were incubated under standard conditions, except for embryonic days 10–18, during which they were incubated at 39°C and 65% RH for 18 h daily. On the 7th day of incubation, eggs with dead embryos were excluded. After hatching was complete, each group was further subdivided into saline-treated or E. coli-challenged groups. The E. coli (serotype 078 with the dose of 1.5 × 105 colony-forming unit/mL) challenge was performed when the birds were 20 days old. Body weight and Tb measurements were taken on post-hatch days 20, 21, 23, and 25. Splenic mRNA expression of cytokines (IL-1β, IL-2, IL-6, IL-8, IL-12, IL-15, IL-16, IL-18, and IFN-γ) was analyzed by real-time quantitative polymerase chain reaction. Following the E. coli challenge, the TM-treated group’s body performance parameters (BW and Tb) were significantly increased compared with the control group. Body weight was higher in the TM group than in the control group (p < 0.05); Tb was lower in the TM group than in the control group (p < 0.05). The mRNA levels of IL and IFN-γ were more stable and moderately induced in the TM group compared with the control group. Thermal manipulation altered the basal mRNA levels of ILs and IFN-γ and changed their expression dynamics after the E. coli challenge. Thermal manipulation during embryogenesis could boost the immune system response to E. coli. Keywords: broiler, challenge, Escherichia coli, immune response, incubation, thermal manipulation.
Research Article | 07 May 2023
Effects of ultrasound-induced stress on gut microbiota of mice
Irina Chernukha, Ekaterina Vasilevskaya, Ksenia Klimina, Roman Yunes, Nadezhda Kupaeva, Galina Tolmacheva, Anastasiya Kibitkina, Valery Danilenko, Sergey Karabanov, and Liliya Fedulova

Volume-16 | Issue-5 | Article-5 | https://doi.org/10.14202/vetworld.2023.929-938

Preview Abstract
Prolonged stress causes deleterious effects on both the organism and its microbiota. In this study, we examined the effects of exposure to variable frequency ultrasound (US) on the gut microbiota-liver-brain axis of mice. This study was conducted on 20 mature clinically healthy sexually naive C57BL/6J male mice (42–45 days old). Group 1 (Normal) consisted of healthy intact mice (n = 10). Group 2 (Stress) consisted of mice subjected to US-induced stress (n = 10) for 20 days with alternating frequencies (20–45 kHz). Stool samples were collected on days 0, 10, and 20, and the corresponding DNA was later subjected to 16SrRNA sequencing. After mice were sacrificed on day 21, the leukocyte count, blood serum biochemical parameters, and liver and brain antioxidant status were measured. Behavioral testing was performed on days 17, 18, and 19. Ultrasound lead to higher stress and anxiety levels; increase in creatinine by 8.29% and gamma-glutamyltransferase activity by 5 times, a decrease in alkaline phosphatase activity by 38.23%, increase of de Ritis coefficient by 21.34%; increased liver and brain superoxide dismutase level by 20.8% and 21.5%, respectively; the stress-related changes in the gut microbiota composition – Bacteroidaceae and Firmicutes. Subjecting mice to 20 days of US-induced stress leads to systemic disorders due to oxidative stress and a decrease in the diversity of the gut microbiota. Keywords: antioxidant system, bacterial metabolites, gut microbiota, ultrasound-induced stress.

Volume-16 | Issue-5 | Article-6 | https://doi.org/10.14202/vetworld.2023.939-945

Preview Abstract
Salmonellosis is a significant foodborne disease that causes serious illness in the gastrointestinal of humans and it is a public health problem worldwide. This study aimed to determine Salmonella spp. in animal feeds, its characteristic, serovar identification, genotyping, and drug sensitivity. Salmonella spp. from animal feedstuffs was collected from January 1 to December 31, 2017, with 657 samples. Serogroup classification was performed by using the slide agglutination test. Then, the samples were analyzed for genotype patterns using pulsed-field gel electrophoresis (PFGE) for DNA fingerprint and antibiotic sensitivity by Vitek® 2 techniques. A total of 80 samples (12.17 %) were found to be Salmonella positive; commercial feed compounds of 60 samples (75%); soybean meal of 10 samples (12.5%); pork meal of 5 samples (6.25%); a fish meal of three samples (3.75%) and poultry meal of 2 samples (2.5%). Serogroups B, C, D, and E were found in this study; 8 samples (10%), 39 samples (48.75%), 8 samples (10%), and 13 samples (16.25%), respectively. A total of 12 samples (15%) were not determined by serogrouping. The most common serovars were Salmonella Rissen (10%), Salmonella Mbandaka (8.75%), and Salmonella Livingstone (6.25%), which belong to serogroup C. Nine of eleven pulsotypes were detected when analyzed by PFGE, showed similarity index between 40.8 and 100 %. Antimicrobial susceptibility tests by Vitek® 2 compact for 11 strains were classified into three groups: resistance to 4, 8, and 11 antibiotics, out of 20 antibiotics. This study revealed annual variation of Salmonella spp. Serovar, genotype, and phenotype from commercial feed compounds and raw materials of which involved people must be aware. Keywords: commercial feed compounds, genotype, Salmonella spp., serotype.
Research Article | 09 May 2023
Global commercialization and research of veterinary vaccines against Pasteurella multocida: 2015–2022 technological surveillance
Aníbal Domínguez-Odio and Daniel Leonardo Cala Delgado

Volume-16 | Issue-5 | Article-7 | https://doi.org/10.14202/vetworld.2023.946-956

Preview Abstract
Pasteurella multocida can infect a multitude of wild and domesticated animals, bacterial vaccines have become a crucial tool in combating antimicrobial resistance (AMR) in animal production. The study aimed to evaluate the current status and scientific trends related to veterinary vaccines against Pasteurella multocida during the 2015–2022 period. The characteristics of globally marketed vaccines were investigated based on the official websites of 22 pharmaceutical companies. VOSviewer® 1.6.18 was used to visualize networks of coauthorship and cooccurrence of keywords from papers published in English and available in Scopus. Current commercial vaccines are mostly inactivated (81.7%), adjuvanted in aluminum hydroxide (57.8%), and designed to immunize cattle (33.0%). Investigational vaccines prioritize the inclusion of attenuated strains, peptide fragments, recombinant proteins, DNA as antigens, aluminum compounds as adjuvants and poultry as the target species. Despite advances in genetic engineering and biotechnology, there will be no changes in the commercial dominance of inactivated and aluminum hydroxide-adjuvanted vaccines in the short term (3–5 years). The future prospects for bacterial vaccines in animal production are promising, with advancements in vaccine formulation and genetic engineering, they have the potential to improve the sustainability of the industry. It is necessary to continue with the studies to improve the efficacy of the vaccines and their availability. Keywords: adjuvant, Pasteurella multocida, strain, technological surveillance, vaccine.
Research Article | 09 May 2023
Factors associated with pet dog behavior in Thailand
Worakan Boonhoh, Tuempong Wongtawan, Prarom Sriphavatsarakom, Natalie Waran, and Chaiwat Boonkaewwan

Volume-16 | Issue-5 | Article-8 | https://doi.org/10.14202/vetworld.2023.957-964

Preview Abstract
Dog behavior problems pose serious public health and economic and animal welfare concerns. There are many factors influencing dog behavior. This study aims to explore factors associated with pet dog behavior in Thailand using the Canine Behavioral Assessment and Research Questionnaire (C-BARQ). The Thai version of the C-BARQ was validated in 2022. The online C-BARQ survey (Google Forms) was advertised and distributed on social media for 3 months. There were a total of 1827 respondents to the survey. The relationship among 12 subscales and variables was analyzed using Spearman’s correlation. “Excitability,” “attachment- and attention-seeking,” and “chasing” were the three most reported behavior problems. “Trainability,” a favored dog behavior, also had a high score in the study. These behaviors were associated with the owner’s age, house type, the dog’s historical background, the number of dogs and the presence of other species in the household, the dog’s training, diet types, and the daily number of meals. The lowest mean score was for “owner-directed aggression,” and it was associated with the dog’s gender and size, the number of dogs and the presence of other species in the household, the dog’s training, and the number of daily meals. This is the first empirical study demonstrating factors related to dog behavior in Thailand. It provides an in-depth understanding of the prevalence and factors associated with Thai pet dog behavior and important knowledge for further studies to advocate for dog–human relationships and contribute to a reduction in dog abandonment in Thailand. Keywords: canine behavioral assessment and research questionnaire, dog behavior, questionnaire, dog-human relationships, Thai dogs.
Review Article | 11 May 2023
Effect of dietary garlic (Allium sativum) on the zootechnical performance and health indicators of aquatic animals: A mini-review
Daniel Leonardo Cala Delgado, Linda Lucia Castillo Caceres, Sebastián Alexander Cely Gómez, and Aníbal Domínguez Odio

Volume-16 | Issue-5 | Article-9 | https://doi.org/10.14202/vetworld.2023.965-976

Preview Abstract
Considerable efforts have been made by modern aquaculture to mitigate the environmental damages caused by its practices while also attempting to improve the quality of the aquatic organisms by promoting alternatives, such as the use of natural products, like garlic (Allium sativum), and instead of chemical agents. Garlic has multiple properties, including antifungal, antibacterial, antiviral, antitoxic, and anticancer effects. In fish, the antiparasitic activity of garlic is one of the most reported effects in the literature, mainly using immersion baths for aquatic organisms. Using garlic also has an antimicrobial effect on the culture of aquatic organisms. Therefore, this review focuses on the impact of garlic on the health and production of aquatic organisms.
Research Article | 11 May 2023
The global profile of antibiotic resistance in bacteria isolated from goats and sheep: A systematic review
Okti Herawati, Siti Khairani Bejo, Zunita Zakaria, and Siti Zubaidah Ramanoon

Volume-16 | Issue-5 | Article-10 | https://doi.org/10.14202/vetworld.2023.977-986

Preview Abstract
Antibiotic resistance has become an issue of global importance due to increasing levels of bacterial infections worldwide. Farm management and usage of antibiotics in livestock are known risk factors associated with the increase in global levels of antibiotic resistance. Goats and sheep are examples of livestock with large populations. Although antibiotic resistance in bacteria from livestock negatively affects both human health and the economy, the global data regarding this issue in goats and sheep are limited. Therefore, this study aimed to provide information on the antibiotic-resistance profile of bacteria isolated from goats and sheep worldwide (Asia, Europe, and Africa). We performed a systematic review of articles published on this topic without any restriction on the year of publication. We searched the Directory of Open Access Journals, PubMed, Google Scholar, and Scopus using Boolean logic through various keywords. The search generated a total of 1325 articles, and after screening for duplicates and implementing inclusion and exclusion criteria, qualitative synthesis (i.e., qualitative systematic review) was performed on 37 articles. The synthesized information indicated that 18 Gram-positive and 13 Gram-negative bacterial species from goats and sheep were resistant to ten antibiotics, namely penicillin, ampicillin, amoxicillin, chloramphenicol, streptomycin, tetracycline, cephalothin, gentamicin, ciprofloxacin (CIP), and sulfamethoxazole. The prevalence of antibiotic resistance ranged from 0.4% to 100%. However, up to 100% of some bacteria, namely, Salmonella Dublin, Aeromonas caviae, and Aeromonas sobria, were susceptible to CIP. Staphylococcus aureus and Escherichia coli were highly resistant to all antibiotics tested. Moreover, eight of the ten antibiotics tested were critically important antibiotics for humans. Antibiotic-resistant bacteria in goats and sheep are a potential risk to animal and human health. Collaboration between all stakeholders and further research is needed to prevent the negative impacts of antibiotic resistance. Keywords: antibiotic, bacteria, goat, resistance, sheep.
Review Article | 13 May 2023
The effect of using fruit peel on broiler growth and health
Sugiharto Sugiharto

Volume-16 | Issue-5 | Article-11 | https://doi.org/10.14202/vetworld.2023.987-1000

Preview Abstract
To ensure the long-term viability of broiler farming, producers must address a number of issues, including rising feed costs, a ban on antibiotic growth promoters, and growing consumer awareness of chemical residues in broiler chicken meat. Fruit peel is a waste with no commercial value, but due to its high nutritional content, particularly in terms of energy, it has the potential to be used as an alternative feed source for broiler chicks. Fruit peel also contains a number of nutraceutical compounds that have the potential to be added to feed or used as natural supplements for broiler chickens due to their antibacterial, antioxidant, and immunostimulant properties. Fruit peels have high fiber content and antinutritional and toxic components that may interfere with broiler digestion and physiological function, so they should be used cautiously in broiler production. Various processes, including fermentation, extraction, distillation, and combining with other active components, such as enzymes, may be used to optimize the use of fruit peels in broiler production. This review examines the use of fruit peel and its effects on broiler growth and health.
Research Article | 13 May 2023
Prevalence of multidrug-resistant and extended-spectrum β–lactamase-producing Escherichia coli from chicken farms in Egypt
Gamal A. Salem, El-Sayed A. Abdelaziz, Mohammed A. Kamel, Nasreddin R. Rhouma, and Reem I. Ali

Volume-16 | Issue-5 | Article-12 | https://doi.org/10.14202/vetworld.2023.1001-1007

Preview Abstract
Extended-spectrum β-lactamase (ESBL)-producing Escherichia coli strains exhibit antibiotic resistance and are known to infect humans worldwide. This study assessed the phenotypic and genotypic prevalence of ESBL-resistant E. coli isolates recovered from the respiratory tracts of chickens in El-Sharkia Governorate, Egypt. We obtained 250 lung samples (one lung/bird) from 50 chicken farms (5 chickens/farm) to isolate, identify, and serotype E. coli. Antimicrobial resistance susceptibility was determined using the disk diffusion method, while the ESBL phenotype was identified using double disk synergy. We detected the β-lactamase genes, blaTEM, and blaSHV, using a polymerase chain reaction. The results showed that 140/250 (56%) were infected with E. coli. All the serogroups of isolated E. coli exhibited high multi-antimicrobial resistance index values (>0.2), and 65.7% were confirmed to have ESBL. Among the isolates with the ESBL phenotypes, 55 (60%) and 32 (35%) contained the blaTEM and blaSHV genes, respectively. The widespread distribution of multidrug-resistant and ESBL-producing E. coli among poultry farms is a significant human health hazard. These results will help the Egyptian authorities to implement a national one-health approach to combat the antimicrobial resistance problem. Keywords: antimicrobial resistance, β-lactamase, chicken farms, Escherichia coli.
Research Article | 13 May 2023
Histopathology and virulence of an in vitro-adapted Trypanosoma evansi TEDC 953 strain (Thailand isolate) in mice
Wallaya Phongphaew, Charuwan Wongsali, Thanisorn Boonyakong, Theerawat Samritwatchasai, Wissanuwat Chimnoi, and Ketsarin Kamyingkird

Volume-16 | Issue-5 | Article-13 | https://doi.org/10.14202/vetworld.2023.1008-1017

Preview Abstract
Trypanosoma evansi is a blood and tissue protozoan parasite affecting domestic and wild animals. The T. evansi Thai strain, namely, T. evansi from dairy cattle number 953 (TEDC 953) strain, has been successfully isolated from dairy cattle and cultivated in vitro. The in vitro-cultivated parasite is useful for biological studies, evaluation of novel chemotherapeutic agents, and production of antigens for diagnostic tests. This study aimed to observe the histopathology and virulence of an in vitro-adapted T. evansi TEDC 953 strain in vivo. The histopathology and virulence of the TEDC 953 strain were clarified in mice. Six mice were infected with 1 × 105 trypomastigotes of TEDC 953 strain intraperitoneally, and four mice were in the negative control. Parasitemia was monitored daily, and the mice were euthanized on 30 days post-infection (DPI). Internal organs were collected for histopathological examination using hematoxylin and eosin staining. Histopathological lesions were found in the liver, lung, heart, kidney, spleen, and brain of the inoculated mice. The main histopathological feature was lymphoplasmacytic inflammation in parenchyma and perivascular areas of multiple organs, and the severity of histopathological changes was related to the presence of trypomastigotes in the regional vessels. Granulomatous inflammation was seen in meninges, pleura, renal capsule, renal pelvis, and spleen of some infected mice. Four mice died at 17, 24, 26, and 27 DPI with an average parasitemia of 4.05 × 1011 trypomastigotes/mL. The average survival time was 23.5 DPI (mice = 4). This study confirmed that the TEDC 953 strain is infectious and pathogenic in mice after the continuously cultivated in vitro. To replace the use of experimental animals, the in vitro-cultivated parasite can be used instead in further studies. Keywords: experimentally infected mice, histopathology, in vitro, in vivo, Trypanosoma evansi Thai strain, virulence.
Research Article | 13 May 2023
Antifungal activity of protein hydrolysates from Thai Phatthalung Sangyod rice (Oryza sativa L.) seeds
Juthatip Jeenkeawpieam, Prawit Rodjan, Sittiruk Roytrakul, Kritsada Pruksaphon, Watcharapong Mitsuwan, Noppharat Tanthanathipchai, Chaiwat Boonkaewwan, Irma Tedja, and Monsicha Pongpom

Volume-16 | Issue-5 | Article-14 | https://doi.org/10.14202/vetworld.2023.1018-1028

Preview Abstract
Fungal zoonoses are an economic and public health concern because they can cause various degrees of morbidity and mortality in animals and humans. To combat this issue, alternative natural antifungals, such as products derived from rice protein hydrolysates or rice antifungal protein/peptide are being considered because they are highly bioactive and exhibit various functional properties. Thailand is a leading rice producer and exporter. Among the various cultivated rice varieties, Sangyod rice (Oryza sativa L.) is exclusively indigenous to Thailand’s Phatthalung province; it has a Thai geographical indication tag. Here, we investigated whether the Phatthalung Sangyod rice seeds have bioactive antifungal peptides. Antifungal activity in four Sangyod rice seed extracts (SYPs) – namely, (1) the crude lysate, SYP1; (2) the heat-treated lysate, SYP2; (3) the heat- and pepsin digested lysate, SYP3; and (4) the heat- and proteinase K-digested lysate, SYP4 – was analyzed. Protein concentrations in these SYPs were determined using the Bradford assay. The total phenolic compound content was determined using the modified Folin–Ciocalteu method in a 96-well microplate. Then, the SYP protein pattern was determined using the sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Subsequently, using the agar well diffusion method, the antifungal properties of these SYPs were tested against ten medically important pathogenic fungi. The minimal inhibitory concentration (MIC) and minimal fungicidal concentration values were determined for the active SYPs – SYP2-4. Finally, the clinical safety of SYP4 was determined using a hemolytic assay (using canine red blood cells [RBCs]). The crude lysate SYP1 did not show antifungal activity against any of the ten tested pathogenic fungi. Surprisingly, hydrolysates SYP2, SYP3, and SYP4 displayed antifungal properties against the ten tested pathogenic fungi. Thus, heat and enzymatic hydrolysis seem to transform the bioactivity of the crude protein extract – SYP1. Further, SYP4 shows the most effective antifungal activity. It completely inhibited Cryptococcus neoformans, Talaromyces marneffei yeast phase, Trichophyton mentagrophytes, and Trichophyton rubrum. A partial inhibitory action on Candida albicans and Microsporum gypseum was possessed while showing the least activity to C. neoformans. SYP4 was nontoxic to canine RBCs. Hemolysis of canine RBCs was undetectable at 1 × MIC and 2 × MIC concentrations; therefore, it can be safely used in further applications. These results indicate that heat and proteinase K hydrolyzed SYP is a very potent antifungal preparation against animal and human fungal pathogens and it can be used in future pharmaceuticals and functional foods. Keywords: antifungal peptide, Oryza sativa L., Phatthalung Sangyod rice, protein hydrolysate, Thai rice seed, zoonoses.
Research Article | 15 May 2023
First molecular detection of the presence of honey bee viruses in insects, Varroa destructor mites, and pollinated plants in an isolated region of Armenia
Hranush Arzumanyan, Hranush Avagyan, Henry Voskanyan, Liana Simonyan, Jon Simonyan, Zara Semirjyan, and Zaven Karalyan

Volume-16 | Issue-5 | Article-15 | https://doi.org/10.14202/vetworld.2023.1029-1034

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Recently, viral diseases of honey bees (Apis mellifera) have presented an increasing threat to beekeeping. This study aimed to examine the presence of honey bee viruses in Apis and non-Apis bee species, the mite Varroa destructor, and pollinated plants in Armenia. Sampling was performed in Tavush Province, in the northeast of the Republic of Armenia, from August to November 2019. Overall, 200 A. mellifera bees, 50 V. destructor mites, and 20 wasps were collected (corresponding to three bees, five mites, and 2–11 wasps in each investigated sample) and homogenized for RNA isolation and detection of viruses. Ten pollinated plants were taken from each plant, and 2 g of each sample was used for homogenization. In each investigated case Apis mellifera, Varroa destructor, Vespula germanica and plants received percentages of the virus presence. Six important honey bee viruses (acute bee paralysis virus [ABPV], deformed wing virus [DWV], A. mellifera norovirus [ANV], Lake Sinai virus-2 [LSV-2], Big Sioux River virus [BSRV], and A. mellifera filamentous virus [AmFV]) were detected in samples by polymerase chain reaction. Our results showed that DWV, ANV, and ABPV were the most common viruses in honey bees. All viruses were detected in wasps, but LSV-2 and ANV were present in almost all samples. Our results showed that almost all viruses were present in V. destructor. Although ANV is very common in honey bees, it did not appear in any mite samples. Our study indicates that viruses typically associated with honey bees were also actively infecting wasps. Our data suggest that the survival of viruses in plants can be an important source of seasonal transmission of viruses to bees. In addition, pollinated plants can potentially serve as reservoirs for honey bee viruses. Keywords: Apis mellifera, honey bee virus, polymerase chain reaction assay, pollinated plants, Varroa destructor.
Research Article | 15 May 2023
In-feed nutritional additive probiotic Saccharomyces boulardii RC009 can substitute for prophylactic antibiotics and improve the production and health of weaning pigs
Julián Parada, Alejandra Magnoli, Maite Corti Isgro, Valeria Poloni, Analía Fochesato, María Pía Martínez, Alicia Carranza, and Lilia Cavaglieri

Volume-16 | Issue-5 | Article-16 | https://doi.org/10.14202/vetworld.2023.1035-1042

Preview Abstract
Non-therapeutic antibiotic use is associated with the current decrease in antibiotic therapeutic efficiency and the emergence of a wide range of resistant strains, which constitutes a public health risk. This study aimed to evaluate the use of Saccharomyces cerevisiae var. boulardii RC009 as a nutritional feed additive to substitute the prophylactic use of antibiotics and improve the productive performance and health of post-weaning piglets. Four regular nutritional phases were prepared. Post-weaning pigs (21-70 days old) received one of two dietary treatments: T1-basal diet (BD-control group) with in-feed antibiotics as a prophylactic medication (one pulse of Tiamulin in P3 and one pulse of Amoxicillin in P4); and T2-BD without in-feed antibiotics but with Saccharomyces boulardii RC009 (1 × 1012 colony forming unit/T feed). The feed conversion ratio (FCR), total weight gain (TWG-kg), and daily weight gain (DWG-kg) were determined. A post-weaning growth index (GI) was calculated and animals (160 days old) from each treatment were analyzed at the abattoir after sacrifice for carcass weight and respiratory tract lesions. Pigs consuming probiotics had higher TWG and DWG than the control group. The group of animals with low body weight obtained the same results. Saccharomyces boulardii administration decreased diarrhea, and FCR reduction was related to a GI improvement. A significant increase in carcass weight and muscle thickness reduction was observed in animals received the probiotic post-weaning. Saccharomyces boulardii RC009, a probiotic additive, was found to improve the production parameters of pigs post-weaning and enhance their health status, indicating that it may be a promising alternative to prophylactic antibiotics. Keywords: nutritional feed additive, post-weaning pig, prophylactic antibiotics, Saccharomyces cerevisiae var. boulardii RC009.
Research Article | 17 May 2023
Gastrointestinal helminth parasites of pets: Retrospective study at the veterinary teaching hospital, IPB University, Bogor, Indonesia
Yusuf Ridwan, Etih Sudarnika, Tri Isyani Tungga Dewi, and Novericko Ginger Budiono

Volume-16 | Issue-5 | Article-17 | https://doi.org/10.14202/vetworld.2023.1043-1051

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Dogs and cats are popular pets that play integral roles in human societies worldwide. Unfortunately, they can carry potential zoonotic helminths that can be transmitted to humans. However, data on the gastrointestinal helminths affecting dogs and cats in Bogor, Indonesia, are currently lacking. This study aimed to investigate the occurrence of gastrointestinal helminth parasites in pets from this area using a retrospective analysis. A retrospective study was conducted at the Veterinary Teaching Hospital of IPB University, Bogor. Cat and dog patients from January 2014 to April 2019 were tested for helminth infections and the results as well as their age, sex, and breed data were analyzed using the Chi-square test. Among the dogs and cats examined for internal parasites, 61.11% (33/51) of the dogs and 53.80% (92/171) of the cats were infected by helminths. Among the dogs, hookworm (37.04%) and Toxocara spp. (24.07%) were detected, while in the cats, hookworm (11.11%), Toxocara spp. (38.01%), and Dipylidium caninum (4.68%) were detected. The prevalence of hookworm and D. caninum was higher in older pets, while Toxocara spp. infected younger cats and dogs (<1 year) more frequently. The prevalence of Toxocara spp. in the Indonesian local dog breed was higher when than other breeds. Sex did not significantly affect the prevalence of parasites in dogs or cats. The discovery of zoonotic helminth parasites in the cat and dog pets from Bogor raises concerns for the inhabitants. Initiatives will be required to inform pet owners about prevention strategies for these parasitic diseases. Keywords: Bogor, cat, dog, gastrointestinal helminth, zoonosis.
Research Article | 17 May 2023
Bird immunobiological parameters in the dissemination of the biofilm-forming bacteria Escherichia coli
Ekaterina Lenchenko, Nadezhda Sachivkina, Tatiana Lobaeva, Natallia Zhabo, and Marina Avdonina

Volume-16 | Issue-5 | Article-18 | https://doi.org/10.14202/vetworld.2023.1052-1060

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With the development of industrial maintenance technology, a group of pathogens called avian pathogenic Escherichia coli (APEC) became very common. The initiation, development, and outcome of the infectious process mediated by virulent APEC strains occur through a decrease in the colonization resistance of the intestine, an immunobiological marker of homeostasis stability in susceptible species. This study focused on the pathogenetic features of colibacillosis and the morphological features of E. coli. Clinical, immunological, bacteriological, and histological studies were conducted on 15-day-old white Leghorn birds (n = 20). The birds were divided into two groups: Control group (Group I; n = 10) with birds intranasally inoculated with 0.5 mL of 0.9% NaCl solution and experimental group (Group II; n = 10) with birds intranasally inoculated with 0.5 mL of an E. coli suspension at 1 billion/mL. During the biofilm formation, clusters of microcolonies were formed as a gel-like intercellular matrix that accumulated due to cell coagulation. The intercellular matrix “glues” heteromorphic cells together and forms a structure of densely packed heteromorphic cells arranged in an orderly manner and growing in different directions. During the experimental reproduction of E. coli, excessive growth was observed in material isolated from poultry. Pathogenic E. coli strains implementing virulence factors adhered to the receptors of erythrocytes, alveolocytes, and enterocytes. Multicellular heterogeneous biofilms, united by an intercellular matrix, were located at the apical poles of the respiratory tract alveolocytes and enterocytes of the terminal ileum villi. Many bacteria exudate containing desquamated epithelial cells with an admixture of mucus, and polymorphonuclear leukocytes were detected in the lumen of the birds’ abdominal organs. Invasive bacteria damaged the epithelial layer, violated the endothelial layer of blood vessels, and developed inflammatory hyperemia of the lamina propria of the respiratory and digestive systems’ mucous membrane. A correlative dependence of changes developed by the type of delayed hypersensitivity reaction was established. Signs of accidental transformation of the thymus, atrophy of the bursa of Fabricius, disseminated thrombosis, and septic spleen developed. Moreover, toxic cardiomyocyte dystrophy, signs of congestive vascular hyperemia, massive disintegration of lymphocytes, macrophage reactions, perivascular edema resulting from the release of plasma, and shaped blood elements were detected. The development and outcome of the infectious process in escherichiosis primarily depend on the homeostasis stability of susceptible species and virulence factors of the pathogenic microorganisms. One of the selected strains, E. coli O78:K80 displayed the highest ability to form biofilms. Its strong adhesion ability to bird erythrocytes was demonstrated. Deepening the scientific knowledge of the interaction between eukaryotes and prokaryotes will contribute to a better understanding of the pathogenetic aspects of avian escherichiosis and eventually find promising anti-adhesive drugs that could reduce primary bacterial contamination in vivo and in vitro. Keywords: avian colibacillosis, biofilm formation, Escherichia coli, intercellular matrix, poultry.

Volume-16 | Issue-5 | Article-19 | https://doi.org/10.14202/vetworld.2023.1061-1070

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Infectious bursal disease (IBD) is an infectious immunosuppressive disease that affects young chickens. Instead of strict biosecurity practices, vaccination is used to control IBD. However, the disease has not been effectively managed. Variations in the observed clinical symptoms lead to confounding diagnoses. The study aimed to obtain pathological lesion data from chickens suspected of IBD virus (IBDV) infection by gross pathology, confirm IBDV infection through molecular diagnostics, and genotype the VP1 gene fragments of circulating IBDV in the field. The bursa of Fabricius, thymus, spleen, proventricular–ventricular junction, thigh muscles, and kidneys samples were collected from chickens suspected of IBDV infection from four commercial broiler farms in Central Java and The Yogyakarta Special Region Province between 2021 and 2022. The collected samples were examined histopathologically. Infectious bursal disease virus RNA was extracted from the bursa of Fabricius and VP1 gene was identified by reverse-transcriptase polimerase chain reaction (RT-PCR). The RT-PCR positive sample were sequenced and analyzed in Mega X for homology search and phylogenetic tree analysis. Macroscopic pathological lesions in the bursa of Fabricius were demonstrated by enlarged edema and thickened plica, presence of gelatinous exudate, hemorrhage, atrophy, and caseous exudate in the lumen. Moreover, the thymus had atrophy and small gray foci were observed in the spleen. Petechiae or hemorrhage was detected on the thigh muscle, and the kidney was dull and pale. Hemorrhage in the proventricular–ventricular junction was distinct. The histopathological examination of the bursa of Fabricius showed follicular vacuolization, edema, heterophilic infiltration, follicular atrophy, congestion, and hemorrhage. The thymus and spleen showed the presence of multifocal necrosis. Hemorrhage was observed in thigh muscle and mucosal part of proventricular–ventricular junction. Vacuolization was seen in renal tubules (nephrosis). Reverse transcriptase-PCR of 26 bursa of Fabricius samples from chickens suspected of IBDV infection showed four negative and 22 positive samples. Phylogenetic analysis of the VP1 gene fragment has indicated very virulent IBD (vvIBD) and belonged to B2 genotype. Infectious bursal diseases virus infection in broiler chicken generated macroscopic and microscopic primary lesions in the bursa of Fabricius and thigh muscle. Other organs such as the spleen, thymus, proventricular–ventricular junction, and kidney, were also involved. Molecular analysis of the VP1 gene confirmed the causative agent and grouped the virus into vvIBD and B2 genotype. All samples were collected from vaccinated birds therefore, the efficacy of available vaccine is required for urgent evaluation. Since most studies only focused on VP1, further exploration on VP2 gene is suggested notably for new-generation vaccines. Monitoring clinical signs’ transformation over time could assist field diagnostics. Keywords: amplification, genotype, infectious bursal disease, pathological lesions.
Research Article | 17 May 2023
Parasitological detection of Theileria parasites in livestock in various regions of the Northern Emirates of the United Arab Emirates
Sameera M. A. Ismaeil, Rajeesh K. T. Parambil, Manal S. Kerab, Ali M. K. ElBakri, and Ideisan I. Abu-Abdoun

Volume-16 | Issue-5 | Article-20 | https://doi.org/10.14202/vetworld.2023.1071-1074

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Theileriosis, caused by tick-borne hemoprotozoans of the genus Theileria, severely impacts the economics of the livestock industry in most tropical and subtropical countries. The aim of the present study was to detect Theileria spp. in domesticated animals (camels, cows, sheep, and goats) using direct microscopy and to determine the infection rate in geographically distinct regions in the northern emirates of the UAE. Blood samples (n = 536) were collected from clinically symptomatic and asymptomatic domesticated animals and subjected to Giemsa staining and examined microscopically for the identification of Theileria. Smears showed an overall rate of positivity for Theileria spp. in 325/536 (60.6%) animals. Different infection rates were recorded across the various animal groups in the different study areas (Middle region 215/386 [55.7%], East region 100/139 [71.9%]). Of the 11 goat samples collected from the North region, 10 (90%) were positive. Infection rates per animal group based on microscopy were as follows: camels, 3/35 (8.5%); cows, 19/36 (52.7%); goats, 200/303 (66%); and sheep, 103/162 (63.5%). Reverse transcription polymerase chain reaction confirmation of all microscopy-positive samples identified 23/325 (7.1%) results as false-positive. This study clarified that Theileria spp. is present in the Middle (Sharjah, Umm Al Quwain, and Ajman), East, and North regions. This report also confirmed the use of direct microscopy with Giemsa-stained blood films as the method of choice for diagnosing acute infections. Further work is needed to molecularly determine the prevalence and species of Theileria spp. circulating in the different parts of the UAE. Keywords: microscopic examination, Theileria, UAE.
Research Article | 17 May 2023
Effect of modified CEP-3 diluents with aqueous soybean extract on liquid semen quality in Ongole crossbred bull
Dian Ratnawati, Kuswati Kuswati, Aulia Puspita Anugra Yekti, Gatot Ciptadi, Sri Rahayu, and Trinil Susilawati

Volume-16 | Issue-5 | Article-21 | https://doi.org/10.14202/vetworld.2023.1075-1083

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Egg yolk (EY) is commonly used as an extracellular cryoprotectant in semen diluents but has some negative effects. Therefore, this study aimed to assess the potential of lecithin derived from plants, such as soybeans, as an alternative extracellular cryoprotectant and to characterize liquid semen quality of Ongole crossbred bulls using a modified caudal epididymis plasma-3 [CEP-3 (m)] as a base diluent and aqueous soybean extract (ASE). A bull with progressive motility (PM) of fresh semen >70% was used. Two soybean extracts were also used, namely, ASE 1 and ASE 2, obtained by extraction procedures 1 and 2, respectively. The study was conducted using an experimental design with 11 treatments and ten replications, with diluents comprising different levels of ASE 1 and ASE 2, as well as a positive control with 10% EY. The parameters measured were motility (M) and its kinetic parameters, including PM, M, velocity curve linear, velocity straight linear, velocity average pathway, linearity, straightness, wobble, amplitude lateral head beat cross frequency, and hyperactivity using computer-assisted sperm analysis, viability, and spermatozoa abnormalities. The CEP-3(m) diluent formula and ASE 1 at a 30% level maintained the PM of spermatozoa up to day 5 (40.7% ± 16.1%) of cold storage. Meanwhile, the CEP-3(m) diluent formula and ASE 2 could only maintain PM >40% until day 3 (42.1% ± 13.5%) of cold storage at a 30% level. The CEP-3(m) diluent and ASE 1 at a level of 25%–30% supported spermatozoa life (viability) up to day 5 with a value >80% (81.8 ± 3.5; 86.4 ± 2.6). The abnormality value of spermatozoa in various diluents during cold storage on days 0–5 was below 20%. Soybean extracts 1 and 2 can substitute EYs as extracellular cryoprotectants in modified CEP-3 basic diluents. Soybean extract 1 can support the life of spermatozoa up to day 5 but may cause the viscosity and movement of spermatozoa to be hyperactive. Soybean extract 2 can support the life of spermatozoa up to the 3rd day of cold storage and produces progressive (non-rotating) movement patterns. Further, research is recommended with higher levels of extract 2. Keywords: aqueous soybean extract, liquid semen, Ongole crossbred cattle.
Research Article | 24 May 2023
Hydroxyapatite-chitosan composites derived from sea cucumbers and shrimp shells ameliorate femoral bone defects in an albino rat model
Arifia Safira, Cinta Atsa Mahesa Rani, Faisal Fikri, Agus Purnomo, Shafia Khairani, Shekhar Chhetri, Salipudin Tasil Maslamama, and Muhammad Thohawi Elziyad Purnama

Volume-16 | Issue-5 | Article-22 | https://doi.org/10.14202/vetworld.2023.1084-1091

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A bone defect is defined as a critically sized autologous bone and a bone gap. Bone grafting is one of the most commonly used surgical methods to enhance bone regeneration in orthopedic procedures. A composite of collagen, hydroxyapatite (HA), and chitosan (Ch) is suitable as a bone matrix and stimulates ossification. This study aimed to evaluate the use of natural HA-Ch composites derived from sea cucumbers and shrimp shells and quantify the levels of cytokines, polymorphonuclear neutrophils (PMNs), serum liver enzymes, calcium, phosphate, and procollagen type 1 N-terminal propeptide (PINP) in albino rats with femoral bone defects. A total of 48 albino rats with femoral bone defects were divided into 4 groups (n = 12 each): (C–) placebo, (C+) polyethylene glycol, (T1) HA, and (T2) HA–Ch groups. Each group was divided into two subgroups (n = 6 each), with euthanization on 7- and 42-day post-treatment, respectively. Procollagen Type 1 N-terminal propeptide and the cytokines interleukin (IL)-4, IL-6, IL-10, and tumor necrosis factor-alpha were quantified using enzyme-linked immunosorbent assay. Flow cytometry was performed to evaluate PMNs. A clinical chemistry analyzer was used to measure the serum levels of liver enzymes, calcium, and phosphate. There was a significant decrease in the level of IL-6 on 7 days and in the level of IL-10 on 42 days in the HA-Ch group. The level of PMNs also decreased significantly on 7 and 42 days in the HA-Ch group. Regarding serum liver enzymes, alkaline phosphatase (ALP) levels in the HA-Ch group increased significantly on 42 days. Calcium and phosphate levels increased significantly on 7 and 42 days in the HA and HA-Ch groups, and PINP levels increased significantly on 7 and 42 days in the HA-Ch group. The HA-Ch composite derived from sea cucumbers and shrimp shells ameliorated femoral bone defects in albino rats. The HA-Ch composite modulated the levels of IL-6, IL-10, PMNs, ALP, calcium, phosphate, and PINP on 7- and 42-day post-treatment. Keywords: bone defect, chitosan, human and health, hydroxyapatite, sea cucumber, shrimp shell.

Volume-16 | Issue-5 | Article-23 | https://doi.org/10.14202/vetworld.2023.1092-1097

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Canine distemper (CD) caused by the CD virus (CDV) has a high mortality rate that severely affects dog populations and other terrestrial carnivores worldwide. However, the genetics of CDV strains circulating in various regions in Vietnam, especially the Mekong Delta, remains unclear. This study aimed to characterize the molecular status of CDV strains circulating in the Mekong Delta, Vietnam. Ocular/nasal swabs were collected from 550 dogs with clinically suspected CDV infection from veterinary clinics in three Vietnamese provinces. A reverse transcription-polymerase chain reaction on the part of the hemagglutinin gene was performed. A phylogenetic tree was constructed to analyze the relationship between the detected CDV and GenBank sequences. The molecular study demonstrated that 4.18% (23/550) of the dogs were positive for CDV. The clinical findings revealed that the positive dogs exhibited clinical signs of distemper. The phylogenetic analysis indicated that the identified CDV sequences were clustered in the same branch with the genotype Asia-1 and distantly related to the vaccine strains. Notably, the CDV sequences detected in this study were grouped with the sequences previously found in southeast Vietnam; however, they were distant from those found in the north. The present study confirmed the presence of CDV and to the best of our knowledge, highlighted for the first time that the CDV strains circulating in the Mekong Delta of Vietnam belong to the genotype Asia-1. Keywords: canine, distemper virus, hemagglutinin gene, phylogenetic analysis, Vietnam.
Review Article | 24 May 2023
A review of dairy cattle heat stress mitigation in Indonesia
Santiananda Arta Asmarasari, Nurul Azizah, Sutikno Sutikno, Wisri Puastuti, Azhar Amir, Lisa Praharani, Supardi Rusdiana, Cecep Hidayat, Anita Hafid, Diana Andrianita Kusumaningrum, Ferdy Saputra, Chalid Talib, Agustin Herliatika, Mohammad Ikhsan Shiddieqy, and Sari Yanti Hayanti

Volume-16 | Issue-5 | Article-24 | https://doi.org/10.14202/vetworld.2023.1098-1108

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Indonesia is a tropical country with a hot climate. In tropical nations such as Indonesia, heat stress is a key reason for the reduced productivity of dairy cattle. Heat stress is a combination of internal and external stimuli that affects an animal, raises its body temperature, and causes it to react physiologically. Most Indonesian dairy cattle are Friesian Holstein (FH), imported from European nations with a temperate environment with low temperatures in the range of 5°C–25°C. Indonesia has a tropical climate with a high ambient temperature that can reach 34°C during the day and the local relative humidity varies between 70% and 90%. Temperature and humidity are two microenvironment factors that may impact the production and heat release in FH cattle. More than 98% of the entire dairy cattle population in Indonesia is found on Java Island. On Java Island, there are between 534.22 and 543.55 thousand heads of cattle, while the dairy cattle population outside Java Island is just 6.59 thousand heads of cattle. The milk output climbs by an average of 3.34% per year, or approximately 909.64 thousand tons and the average annual growth in whole milk consumption was 0.19 L/capita. Indonesian cow milk output has been unable to keep pace with the country’s increasing demand. This study aimed to review the strategies to mitigate heat stress in FH dairy cattle in Indonesia.

Volume-16 | Issue-5 | Article-25 | https://doi.org/10.14202/vetworld.2023.1109-1113

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QX-like infectious bronchitis virus (IBV) is a highly infectious avian coronavirus that causes respiratory and kidney disease. It is linked to increased mortality and loss of performance in infected chickens worldwide, including Thailand. Thus, a simple and rapid diagnostic method for the diagnosis of QX-like IBV is needed. This study aimed to develop a single-step multiplex reverse transcription-polymerase chain reaction (mRT–PCR) assay to detect and differentiate QX-like IBV from Thai IBV and vaccine strains used in the poultry industry (H120, Ma5, and 4/91). Primer sets specific for QX-like and Thai IBV were designed to target the S1 gene. The specificity of the technique was verified using nine isolates of QX-like IBV, four isolates of Thai IBV, and other avian viral respiratory pathogens. The detection limit was evaluated using a serial ten-fold dilution of QX-like and Thai IBV. The results showed that single-step mRT–PCR could detect QX-like IBV and differentiate it from Thai IBV and the vaccine strains H120, Ma5, and 4/91. The limit of detection of the developed assay was 102.2 embryo infectious dose (EID)50/mL for QX-like IBV and 101.8 EID50/mL for Thai IBV. Interestingly, the developed assay could identify mixed infection by both IBVs in a single sample. The single-step mRT–PCR assay developed in this study can potentially discriminate QX-like IBV from Thai IBV and the vaccine strains H120, Ma5, and 4/91 in a single reaction. It is also suitable for use in all laboratories with access to conventional PCR equipment. Keywords: detection, QX-like infectious bronchitis virus, single-step multiplex reverse transcription-polymerase chain reaction, specificity.
Research Article | 27 May 2023
Ultrasonographic kidney length-to-abdominal aortic diameter for the diagnosis of feline chronic kidney disease: A preliminary study
Kotchapol Jaturanratsamee, Nan Choisunirachon, Kumpanart Soontornvipart, Damri Darawiroj, Naparee Srisowanna, and Chutimon Thanaboonnipat

Volume-16 | Issue-5 | Article-26 | https://doi.org/10.14202/vetworld.2023.1114-1121

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Chronic kidney disease (CKD) is one of the most important diseases in cats. This study aimed to compare the ultrasonographic kidney length-to-abdominal aortic diameter (K/AO) ratio between healthy and CKD cats and investigate the correlation between K/AO and blood results. Fifteen healthy cats and 15 CKD cats were included in this clinically prospective study. All cats were evaluated for radiographic and ultrasonographic K, radiographic K-to-second lumbar length ratio (K/L2), and K/AO, indirect systolic blood pressure and plasma creatinine (Cr), blood urea nitrogen (BUN), and symmetric dimethyl arginine (SDMA). The radiographic and ultrasonographic kidney lengths of CKD were significantly shorter than those of healthy cats (p < 0.05 and p < 0.05, respectively). The average K/L2 and K/AO were significantly lower in CKD than in healthy cats (p < 0.01 and p < 0.001, respectively). The K/AO had a strong negative correlation with plasma Cr (r = −0.7682, p < 0.0001), BUN (r = −0.6175, p < 0.001), and SDMA (r = −0.589, p < 0.001). However, K/L2 had a moderate negative correlation with plasma Cr (r = −0.5866, p < 0.001), BUN (r = −0.4884, p < 0.01), and SDMA (r = −0.5404, p < 0.01). The optimal cutoff value of K/AO (<10.71) had higher sensitivity and specificity than K/L2 for identifying feline CKD. Kidney length-to-abdominal aortic diameter could be a better and more promising parameter than the K/L2 ratio for evaluating kidney size in cats with CKD. Keywords: cats, chronic kidney disease, kidney length-to-abdominal aortic diameter, renal length, ultrasonography.
Research Article | 30 May 2023
Management of bovine brucellosis in organized dairy herds through the identification of risk factors: A cross-sectional study from Karnataka, India
Rajeswari Shome, Krithiga Natesan, Triveni Kalleshamurthy, Chaitra Yadav, Swati Sahay, Somy Skariah, Nagalingam Mohandoss, Obli Rajendran Vinodh Kumar, Bibek Ranjan Shome, and Habibur Rahman

Volume-16 | Issue-5 | Article-27 | https://doi.org/10.14202/vetworld.2023.1122-1130

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Brucellosis is an infectious disease caused by Brucella species. This study aimed to identify the risk factors associated with bovine brucellosis seropositivity in organized dairy farms to control the disease in unvaccinated adult bovine herds in Karnataka, India. In total, 3610 samples (3221 cattle and 389 buffaloes) were subjected to parallel testing using the Rose Bengal plate test and protein G-based enzyme-linked immunosorbent assay, followed by analyses of animal- and farm-level epidemiological datasets to identify the risk factors. The apparent brucellosis prevalence at the animal level was higher in buffaloes (8.2%, 95% confidence interval [CI] = 5.9–11.4) than in cattle (6.1%, 95% CI = 5.3–7.0). In a multivariable logistic model, animals calved 3–5 times (odds ratio [OR] = 2.22, 95% CI = 1.50–3.1, reference [ref]: animals calved <2 times); animals with a history of abortion (OR = 54.73, 95% CI = 33.66–89.02), repeat breeding (OR = 19.46, 95% CI = 11.72–32.25), and placental retention (OR = 13.94, 95% CI = 4.92–39.42, ref: no clinical signs); and dogs on farms (OR = 2.55, 95% CI = 1.48–4.40, ref: absence of dogs); disposal of aborted fetus in open fields (OR = 4.97, 95% CI = 1.93–12.84) and water bodies (OR = 2.22, 95% CI = 1.50–3.1, ref: buried); purchase of animals from other farms (OR = 6.46, 95% CI = 1.01–41.67, ref: government farms); hand milking (OR = 1.98, 95% CI = 1.02–10.0, ref: machine milking); and use of monthly veterinary services (OR = 3.45, 95% CI = 1.28–9.29, ref: weekly services) were considered significant risk factors for brucellosis in organized bovine herds (p < 0.01). The study identified that the animals calved 3–5 times or with a history of abortion/repeat breeding/placental retention, and disposal of aborted fetus in open fields/water bodies as the potential risk factors for bovine brucellosis. These risk factors should be controlled through the implementation of best practices to reduce the brucellosis burden in bovine farms. Keywords: brucellosis, buffalo, cattle, India, risk factors, seroprevalence.
Research Article | 30 May 2023
Development of a microencapsulated probiotic containing Pediococcus acidilactici WU222001 against avian pathogenic Escherichia coli
Watcharapong Mitsuwan, Phirabhat Saengsawang, Juthatip Jeenkeawpieam, Veeranoot Nissapatorn, Maria de Lourdes Pereira, Warangkana Kitpipit, Thotsapol Thomrongsuwannakij, Saranporn Poothong, and Sasi Vimon

Volume-16 | Issue-5 | Article-28 | https://doi.org/10.14202/vetworld.2023.1131-1140

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Probiotics are beneficial microorganisms for humans and animals. In this study, we developed a microencapsulated probiotic with antibacterial activity against avian pathogenic Escherichia coli (APEC). Alignment of the 16S rRNA sequences of the isolate WU222001 with those deposited in GenBank revealed that the isolate was Pediococcus acidilactici with 99.6% homology. This bacterium was characterized as a probiotic based on its tolerance toward in vitro gastrointestinal tract (GIT) conditions, hydrophobicity, and auto-aggregation. The antibacterial activity of the probiotic’s culture supernatant against APEC was investigated using a broth microdilution assay. Pediococcus acidilactici was microencapsulated using sodium alginate and agar with diameters ranging from 47 to 61 µm. Then, physicochemical characteristics and stability of the microcapsules were determined. The isolate was characterized as a probiotic based on its resistance to low pH, bile salts, and pancreatin, with relative values of 79.2%, 70.95%, and 90.64%, respectively. Furthermore, the bacterium exhibited 79.56% auto-aggregation and 55.25% hydrophobicity at 24 h. The probiotic’s culture supernatant exhibited strong antibacterial activity against clinical APEC isolates with minimum inhibitory concentration and minimum bactericidal concentration of 12.5% and 25% v/v, respectively. Microencapsulation-enhanced bacterial viability in GIT compared to free cells. Moreover, 89.65% of the encapsulated cells were released into the simulated intestinal fluid within 4 h. The viable count in microcapsules was 63.19% after 3 months of storage at 4°C. The results indicated that the culture supernatant of P. acidilactici inhibited the growth of APEC. In addition, microencapsulation extends the viability of P. acidilactici under harsh conditions, indicating its potential application in the feed production. Keywords: antibacterial activity, avian pathogenic Escherichia coli, microcapsule, microencapsulation, Pediococcus acidilactici, probiotics.
Research Article | 30 May 2023
Artemisia vulgaris anthelmintic activities to ova and adult stages of Fasciola gigantica in vitro
Andini Nurlaelasari, A’isyah Retno Wulandari, Tamara Muñoz Caro, Herjuno Ari Nugroho, Sukaryo Sukaryo, Muhammad Cahyadi, Wahyu Kurniawan, and Penny Humaidah Hamid

Volume-16 | Issue-5 | Article-29 | https://doi.org/10.14202/vetworld.2023.1141-1153

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Fasciolosis due to Fasciola gigantica is endemic to tropical countries and Fasciola hepatica in temperate climates, highly detrimental to livestock and known as foodborne zoonotic diseases. The strategic control of the disease is mainly the use of chemical anthelmintic. This study aimed to evaluate the anthelmintic properties of Artemisia vulgaris extract on the ova and adult stages of F. gigantica. Samples were collected from the Ampel Abbatoir, Boyolali District, Central Java, Indonesia. The ova from 20-gallbladders of cattle which were naturally infected with F. gigantica and 270 living F. gigantica worms were used in this study. The ovicidal assay was performed by incubating the ova with A. vulgaris in different concentrations, that is, 5%, 2.5%, and 1.25% for 5, 9, 11, 14, and 16 days. The efficacies were evaluated by quantification of ova degeneration during developmental stages in different time points and egg-hatch assay. The flukicidal effects were observed by mortality assay in 5, 10, 20, 40, 80, 160, 320, and 640 min incubations followed by scanning electron microscopy for surface morphology and histology of the fluke’s transversal sections. The concentration of 5% A. vulgaris showed the strongest ovicidal activities. The percentage of hatching ova on day 16 at concentrations of 5%, 2.5%, and 1.25% were 3.33%, 6.67%, and 16.67%. These ova hatch assay showed a significant reduction (p < 0.001) compared to untreated control. The flukicidal effect was significant (p < 0.001) at a concentration of 20%, with a mortality rate reaching 66.67% in the 40 min of incubation time. The surface properties of the adult worms, including the spine, tegument, acetabulum, intestine, and vitelline follicles, were disintegrated. The results showed that A. vulgaris has the potential ovicidal and flukicidal properties to F. gigantica. The active compounds remained necessary to be elucidated further and its modes of action would be interesting to be predicted by molecular docking modeling. Keywords: anthelmintic, Artemisia vulgaris, Fasciola gigantica, flukicidal, ovicidal.
Research Article | 30 May 2023
West Nile virus: The current situation in Egypt
Rabab T. Hassanien, Heba A. Hussein, Hala K. Abdelmegeed, Dina A. Abdelwahed, Omnia M. Khattab, M. H. Ali, Ahmed R. Habashi, Essam M. Ibraheem, Momtaz A. Shahein, and Eman M. Abohatab

Volume-16 | Issue-5 | Article-30 | https://doi.org/10.14202/vetworld.2023.1154-1160

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Due to climatic changes, arthropod-borne viruses have become a global health concern. In Egypt, West Nile virus (WNV) was initially detected in humans in 1950 and then in 1951, 1954, 1968, and 1989. Although WNV infection has been recorded in numerous Middle Eastern countries, its prevalence among the equine population in Egypt is unknown. This study aimed to investigate the current situation of vector-borne WNV in Egypt, estimate its seroprevalence, and assess the associated risk factors. We screened 1100 sera samples and nasal swabs from the same equids, 156 mosquito pools, and 336 oropharyngeal and cloacal swabs from migratory birds for WNV. The sera were investigated for the presence of immunoglobulin G (IgG) and immunoglobulin M (IgM) against WNV-prE. Real-time reverse transcription-polymerase chain reaction was used to detect WNV RNA in the nasal swab samples, mosquito pools, and migratory birds’ oropharyngeal and cloacal swabs. The seroprevalence showed positive IgG in sera samples collected from different districts. The data showed that horses were 1.65-fold more susceptible than donkeys, with male being 1.45 times more susceptible than females. Moreover, the tested equids samples were divided into three groups based on their age: <5 years, 5–10 years, and >10 years. The 5–10- year group was 1.1 and 1.61 times more vulnerable to infection than the <5- and >10 year groups. All the sera samples were negative for IgM. The nasal swabs from equids, oropharyngeal and cloacal swabs from migratory birds, and mosquito samples tested negative for WNV by molecular detection. Based on the obtained data, we recommend that effective control programs should be implemented to enable epidemiological investigations and understand the current situation of WNV in Egypt. Keywords: climatic changes, flaviviruses, seroprevalence, West Nile virus.
Research Article | 30 May 2023
Flow cytometric evaluation of CD4+ and CD8+ T-cell in IPB-D2 chickens with different Newcastle disease antibody titers level
Dwi Lestari, Sri Murtini, Niken Ulupi, Asep Gunawan, and Cece Sumantri

Volume-16 | Issue-5 | Article-31 | https://doi.org/10.14202/vetworld.2023.1161-1164

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IPB-D2 chickens are selected from IPB-D1 due to their disease-resistance characteristics. One-way to evaluate the strength of a chicken’s immune system is by examining the number of circulating T lymphocytes. This assessment can be conducted using a modern analytic method called flow cytometry which relies on monoclonal antibodies to detect the relative proportions of each cell and measure the quality and quantity of biological and physical features of cells, including specific membrane or intracellular glycoprotein markers. Therefore, this study aimed to evaluate the population of lymphocytes, cluster of differentiation (CD)4+ and CD8+ in IPB-D2 chickens. Flow cytometry was used to evaluate the population of lymphocytes, CD4+, and CD8+ in IPB-D2 chickens. The data obtained in this study were analyzed by Minitab, and the mean values were compared using a t-test. The lymphocytes, CD4+, and CD8+ populations of IPB-D2 chicken with high Newcastle disease (ND) antibody titers were 65.04%, 10.53%, and 5.47%. Meanwhile, this breed, with low ND antibody titers had lymphocytes CD4+ and CD8+ population of 57.19%, 8.40%, and 4.11 %. The comparison of CD4+ and CD8+ populations in chickens with high and low ND antibody titers was 1.92 and 2.04, respectively. IPB-D2 chickens with high ND antibody titers exhibited increased lymphocyte, CD4+, and CD8+ cell populations in comparison to those with low ND antibody titers. However, the high ND antibody titer group had a lower CD4+/CD8+ ratio. Keywords: cluster of differentiation 4+, cluster of differentiation 8+, flow cytometry, IPB-D2 chicken, lymphocytes.
Research Article | 31 May 2023
Physicochemical, microbiological, and sensory characteristics of “Sui Wu’u” traditional pork products from Bajawa, West Flores, Indonesia
Yuliana Tandi Rubak, Herianus J. D. Lalel, and Maxs Urias Ebenhaizar Sanam

Volume-16 | Issue-5 | Article-32 | https://doi.org/10.14202/vetworld.2023.1165-1175

Preview Abstract
Sui Wu’u is a traditional meat preservation product from Bajawa, a region in East Nusa Tenggara. It is made by mixing pork with salt and corn flour, which is then stored in a bamboo container (tuku) for months. After 6 months of storage, this study examined the physicochemical, microbiological, and sensory properties of Sui Wu’u. Sui Wu’u products were prepared using the traditional recipe from the Bajawa community. Fresh pork (pork belly and backfat), corn flour, and salt were purchased from local/traditional markets at proportions of 65%, 30%, and 5%, respectively. The physicochemical, amino acid, fatty acid profile, microbiological, and sensory properties of Sui Wu’u were evaluated after being stored for 6 months in a bamboo container (tuku). The results indicated that these Sui Wu’u were mainly characterized by high-fat levels, followed by protein. The pH value, salt content, moisture content, and water activity were 4.72%, 1.72%, 6.11%, and 0.62%, respectively. Minerals (K, P, Se, and Zn) and vitamin B6, as well as amino acids, such as leucine, phenylalanine, lysine (essential amino acids), glycine, proline, glutamic acid, and alanine (non-essential amino acids), are present in Sui Wu’u. The fatty acid profile was dominated by monounsaturated fatty acids (MUFA) (21.69%), saturated fatty acids (SFA) (17.78%), and polyunsaturated fatty acids (PUFA) (5.36%). Monounsaturated fatty acids, oleic acid (C18:1n9) was the most abundant fatty acid in Sui Wu’u, followed by palmitic acid SFA (C16:0); MUFA stearic acid (C18:0); and PUFA linoleic (C18:2n-6). The microbiological characteristics of Sui Wu’u showed no detectable microorganisms (<10 CFU/g) for Salmonella, total E. coli and total Staphylococcus, and average values of 4.4 × 105 CFU/g for total microbes, which were still below the maximum limit of microbial contamination according to the regulations of the Food and Drug Supervisory Agency of the Republic of Indonesia. The sensory assessment indicated that panelists highly preferred (rated as very like) Sui Wu’u for all sensory attributes. The physicochemical, microbiological, and sensory characteristics of Sui Wu’u after 6 months of storage indicated that it still provides essential nutrients for the body and is quite safe for consumption. The stability of Sui Wu’u’s shelf life can be attributed to the appropriate combination of pork, salt, corn flour, bamboo packaging (tuku), and storage temperature. The high-fat content in Sui Wu’u can be reduced by increasing the proportion of lean meat. Ensuring strict sanitation during the manufacturing process, using high-quality pork, salt, corn flour, and proper packaging with bamboo can further improve the safety of Sui Wu’u for consumption. Keywords: Bajawa, meat preservation, sensory characteristics, Sui Wu’u.
Research Article | 31 May 2023
Effectiveness of ozone-laser photodynamic combination therapy for healing wounds infected with methicillin-resistant Staphylococcus aureus in mice
Suryani Dyah Astuti, Wahyu Intan Pertiwi, Sri Puji Astuti Wahyuningsih, Perwira Annissa Dyah Permatasari, Dezy Zahrotul Istiqomah Nurdin, and Ardiansyah Syahrom

Volume-16 | Issue-5 | Article-33 | https://doi.org/10.14202/vetworld.2023.1176-1184

Preview Abstract
According to 2013 data from the Ministry of Health of the Republic of Indonesia, there were 8.2% more wounds than typical in Indonesia; 25.4% were open wounds, 70.9% were abrasions and bruises, and 23.2% were lacerations. A wound is defined as damage or loss of body tissue. This study aimed to determine the effectiveness of wound healing using red-laser therapy (650 nm, 3.5 J/cm2), blue-laser therapy (405 nm, 3.5 J/ cm2), ozone therapy, red-laser therapy (650 nm, 3.5 J/cm2) with ozone, and blue-laser therapy (405 nm, 3.5 J/cm2) with ozone. One hundred and twelve mice were given incision wounds and infected with methicillin-resistant Staphylococcus aureus (MRSA). The study used a factorial design with two factors: The type of therapy (n = 7) and irradiation time (days 1, 2, 4, and 6). The mice were divided into seven therapy groups: Control group with NaCl, control with Sofra-tulle® treatment, red-laser therapy (650 nm, 3.5 J/cm2), blue-laser therapy (405 nm, 3.5 J/cm2), ozone therapy, red-laser therapy (650 nm, 3.5 J/cm2) with ozone, and blue-laser therapy (405 nm, 3.5 J/cm2) with ozone. This therapy was performed using irradiation perpendicular to the wound area. The photosensitizer used was curcumin 10 mg/mL, which was applied to the wound area before exposure to a laser and ozone. The ozone concentration was 0.011 mg/L with a flow time of 80 s. The test parameters were the number of collagens, bacterial colonies, lymphocytes, monocytes, and wound length measurement to determine their acceleration effects on wound healing. Data were analyzed by a two-way (factorial) analysis of variance test. Acceleration of wound healing was significantly different between treatments with a laser or a laser-ozone combination and treatment using 95% sodium chloride (NaCl) and Sofra-tulle®. On day 6, the blue-laser with ozone treatment group had efficiently increased the number of bacteria and reduced the wound length, and the red-laser treatment with ozone increased the amount of collagen. In addition, the red-laser also reduced the number of lymphocytes and monocytes, which can have an impact on accelerating wound healing. Blue-laser therapy was very effective for increasing the number of epithelia. The blue- and red-laser combined with ozone treatments effectively accelerated the healing of incisional wounds infected with MRSA bacteria. Keywords: blue-laser, methicillin-resistant Staphylococcus aureus, ozone therapy, red-laser.