Genetic polymorphisms of the 5' untranslated regions of the HSP70 gene in Indonesian cattle populations

Research (Published online: 27-01-2022)

21. Genetic polymorphisms of the 5' untranslated regions of the HSP70 gene in Indonesian cattle populations

Peni Wahyu Prihandini, Almira Primasari, Aryogi Aryogi, Muchamad Luthfi and Dwi Nur Happy Hariyono

Veterinary World, 15(1): 168-172

ABSTRACT

Background and Aim: Heat shock proteins (HSPs) are a group of proteins that play a significant role in protecting cells against cellular stress. HSP70 is a conserved, sensitive, and abundant gene associated with heat stress's physiological adaptability. The objective of this study was to reveal the polymorphisms of the partial sequences of the HSP70 gene (5' untranslated region [UTR]) in seven cattle populations in Indonesia.

Materials and Methods: Polymerase chain reaction products (551 bp) of the HSP70 gene amplified from 102 animals representing seven cattle populations (Bali, Belgian Blue × Peranakan Ongole [PO] cross, Galekan, Jabres, Madura, PO, and Rambon) were sequenced by DNA sequencing method.

Results: Fourteen single-nucleotide polymorphisms (SNPs), generally found at a low frequency, were detected. Among these SNPs, only 1117G>A, 1125A>C, and 1204T>C were polymorphic in all the analyzed breeds. A Chi-square test showed that the majority of the loci were in Hardy–Weinberg equilibrium (p>0.05). Varying levels of observed (0.050-0.571) and expected heterozygosity (0.049-0.500) were noted. The polymorphism information content values (0.048-0.375) indicated that the SNPs in the HSP70 gene showed low-to-moderate polymorphism in the studied populations. Thirty-six haplotypes were defined according to the identified SNPs, of which haplotype Hap5 (CGACGAGAGTGTCC) and Hap4 (CGACGAGAGTGCCC) were generally dominant in the studied samples. The phylogenetic tree showed a close relationship between Bali and Rambon cattle and between Galekan and Jabres cattle, while the Belgian Blue × PO crossbred cattle were farther apart.

Conclusion: The polymorphisms in the 5' UTR of the HSP70 gene identified in this study should be further investigated in a larger population to unravel the association between the SNPs and thermotolerance in Indonesian local cattle populations.

Keywords: cattle, heat shock protein, heat stress, polymorphism, thermotolerance.

Survivability of Clostridioides difficile spores in fermented pork summer sausage during refrigerated storage

Research (Published online: 27-01-2022)

20. Survivability of Clostridioides difficile spores in fermented pork summer sausage during refrigerated storage

Genevieve Flock, Hsin-Bai Yin, Chi-Hung Chen, Abraham Joseph Pellissery and Kumar Venkitanarayanan

Veterinary World, 15(1): 162-167

ABSTRACT

Background and Aim: Clostridioides difficile is a spore-forming pathogen that causes serious enteric disease in humans. Strains have been isolated from food animals and meat, including pork, which suggest a potential for foodborne transmission. Pork summer sausage is a popular fermented meat product, which is consumed cooked or cooked to a lower internal temperature due to acidification of the product. The effect of acidity and cooking on the viability of C. difficile spores in a fermented meat product has not been determined. Therefore, the aim was to study the survivability of C. difficile spores in fermented pork summer sausage.

Materials and Methods: Fermented pork sausages were prepared according to a commercial recipe with or without starter culture and C. difficile spores followed by fermentation at 37°C for ∼12 h under 85% relative humidity until pH 5.0 was reached and further processed as cooked (>57°C) or uncooked (≤57°C) and stored at 4°C. C. difficile spores in sausages were enumerated at 1 h following inoculation and on days 0, 1, 7, 14, 21, 30, 60, and 90 of storage.

Results: It was observed that C. difficile spore viability in control unfermented treatment was significantly different on day 0 from the fermented, fermented cooked, and control unfermented cooked treatments (p<0.05); however, there was no significant difference among the latter three treatment groups throughout 90 days of storage (p>0.05). On day 90 of storage, the unfermented control sausages yielded ∼4.0 log colony-forming unit (CFU)/g of C. difficile spores compared to ∼3.5 log CFU/g recovered from fermented samples and the unfermented cooked control samples identifying spore viability in all treatment groups.

Conclusion: C. difficile spores were found to survive the acidity and cooking of fermented pork summer sausage and storage at 4°C for 3 months, thereby highlighting the need for effective intervention strategies to reduce the risk of C. difficile contamination in pork products.

Keywords: acidity, Clostridioides difficile, fermented pork sausage, spores.

Effect of trimmed asparagus by-products supplementation in broiler diets on performance, nutrients digestibility, gut ecology, and functional meat production

Research (Published online: 26-01-2022)

19. Effect of trimmed asparagus by-products supplementation in broiler diets on performance, nutrients digestibility, gut ecology, and functional meat production

Manatsanun Nopparatmaitree, Marisa Nava, Verachai Chumsangchotisakun, Pornpan Saenphoom, Soranot Chotnipat and Warangkana Kitpipit

Veterinary World, 15(1): 147-161

ABSTRACT

Background and Aim: Trimmed asparagus by-products (TABP) is the resultant waste from asparagus possessing. TABP has fructans, such as inulins and fructooligosaccharide, which can be utilized as an alternative prebiotic. This study was conducted to examine the effect of TABP dietary supplementation on the productive performance, nutrient digestibility, gut microbiota, volatile fatty acid (VFA) content, small-intestine histology, and meat quality of broilers.

Materials and Methods: A total of 320 1-day-old broiler chicks (Ross 308®) were raised under ambient temperature and assigned through a completely randomized design to one of four dietary treatments, with four replicates per treatment. The dietary treatments comprised corn-soybean basal diet supplemented with 0 (control), 10, 30, or 50 g/kg TABP. All birds were provided drinking water and feed ad libitum to meet the standard nutritional requirements of National Research Council for broiler chickens.

Results: TABP supplementation to the broilers significantly increased the apparent ether extract, crude fiber, and gross energy digestibility (p<0.05). TABP supplementation significantly increased lactic bacteria and Enterococcus spp. numbers as well as acetic, propionic, butyric, and total VFA levels (p<0.01); on the other hand, it also significantly decreased Salmonella spp. and Escherichia coli contents in the cecum compared with the control group (p<0.01). Moreover, TABP supplementation increased villus height in the duodenum and jejunum (p<0.01), cryptal depth in the jejunum and ileum (p<0.01), and villus surface areas in the duodenum, jejunum, and ileum (p<0.01). Overall, 0-35 day TABP supplementation significantly increased the feed intake (p<0.01) and average daily gain of broilers (p<0.05), but not significantly affected the viability, productive index, and economic benefit return (p>0.05). The carcass characteristics, pH, color, and water holding capacity of the chicken meat between groups were not significantly different (p>0.05). All levels of TABP supplementation appeared to be a feasible means of producing broilers with the lower serum low-density lipoprotein cholesterol and triglyceride levels as well as atherogenic indices of serum compared with the control (p<0.05). Cholesterol contents and palmitic acid, oleic acid, saturated fatty acids, and Monounsaturated fatty acids levels decreased with an increase of TABP supplementation (p<0.05). Furthermore, TABP supplementation decreased atherogenic index (AI) and thrombogenicity index (TI) of meat (p<0.05).

Conclusion: Supplementation of 30 g/kg TABP in broiler diet could enhance broiler performance and provide chicken meat with beneficial properties, with decreased AI and TI resulted from altered cholesterol and fatty acid profiles.

Keywords: asparagus by-products, broiler, functional feed, functional meat, prebiotic.

Biochemical and molecular identification of Gram-positive isolates with β-hemolysis activity isolated from the nasal swab of pigs during the human meningitis outbreak in Badung Regency, Bali-Indonesia

Research (Published online: 25-01-2022)

18. Biochemical and molecular identification of Gram-positive isolates with β-hemolysis activity isolated from the nasal swab of pigs during the human meningitis outbreak in Badung Regency, Bali-Indonesia

K. J. Putra Pinatih, I. W. Suardana, I. D. M. Sukrama, I. B. N. Swacita and R. K. Putri

Veterinary World, 15(1): 140-146

ABSTRACT

Background and Aim: The nasal cavity of a pig serves as an entry point and a habitat for the colonization of commensal microbes and pathogenic bacteria. Based on biochemical and serological tests, Streptococcus β-hemolytic Group C was identified as the Gram-positive bacteria, which resulted in the 1994 outbreak and death of thousands of pigs in Bali. Furthermore, this agent is zoonotic and frequently results in the development of meningitis lesions in the infected pig. Recently, a meningitis outbreak in humans was also reported after the consumption of pig-derived foods at Sibang Kaja, Badung-Bali. This study aimed to identify and characterize Gram-positive β-hemolytic organisms collected from nasal swab of pigs from the outbreak area, as well as to compare API Kit and 16S rRNA gene analysis methods.

Materials and Methods: This study commenced with the cultivation of two isolates, Punggul Swab Nasal (PSN) 2 and PSN 19, which were characterized by β-hemolysis activity. These samples were then conventionally and molecularly identified using Kit API 20 Strep and 16S ribosomal RNA (rRNA) gene primers, respectively.

Results: Using the Kit API 20 Strep, both isolates were identified as Enterococcus faecium, which was previously classified as Group D Streptococci. Based on the 16S rRNA gene sequencing, PSN 2 and PSN 19 were molecularly confirmed to have 99 and 98.1% similarities with E. faecium (NR042054), respectively. Furthermore, both isolates share the same clade in the phylogenetic tree analysis.

Conclusion: Using Kit API 20 Strep and 16S rRNA gene analysis, the PSN 2 and PSN 9 Gram-positive isolates with β-hemolysis activity from pig nasal swabs were identified as E. faecium.

Keywords: 16S ribosomal RNA gene, Gram-positive bacteria, Kit API 20 Strep, nasal of pig, phylogenetic tree.

A novel multiplex polymerase chain reaction assay for the genotypic survey of the non-homologous end-joining factor 1 gene associated with Collie eye anomaly in Thailand

Research (Published online: 25-01-2022)

17. A novel multiplex polymerase chain reaction assay for the genotypic survey of the non-homologous end-joining factor 1 gene associated with Collie eye anomaly in Thailand

Chommanad Lerdkrai and Nuch Phungphosop

Veterinary World, 15(1): 132-139

ABSTRACT

Background and Aim: Collie eye anomaly (CEA) is a hereditary and congenital ocular disorder, which affects several dog breeds, including Collies, Collie-related breeds, and other purebreds. An intronic deletion of 7799-bp in the non-homologous end-joining factor 1 (NHEJ1) gene has been identified as the genetic defect causing CEA. This study aimed to investigate the prevalence of CEA based on NHEJ1 genotyping assay in Thailand.

Materials and Methods: We clarified the prevalence of CEA in 224 dogs from five purebred dog breeds using a novel multiplex polymerase chain reaction (PCR)-based technique and confirmed the genotypic status with direct DNA sequencing.

Results: The highest frequency of the mutated NHEJ1 allele was 83.3% for Rough Collies, followed by 7.8% for Border Collies, 5.1% for Australian Shepherds, and 2.8% for Shetland Sheepdogs. The heterozygous mutated NHEJ1 genotype detected for Rough Collies, Border Collies, Australian Shepherds, and Shetland Sheepdogs was 33.3%, 15.6%, 10.3%, and 3.3%, respectively. The homozygous mutated NHEJ1 genotype was detected only in Rough Collies and Shetland Sheepdogs, accounting for 66.7% and 1.1%, respectively. Thai Ridgeback Dogs were not affected by this mutation.

Conclusion: This study describes, for the 1st time, the genotypic survey of the NHEJ1 gene associated with CEA in dogs in Thailand. In addition, we successfully developed a new multiplex PCR assay with high accuracy, reproducibility, and cost-efficiency and validated its usefulness for determining NHEJ1 genotypes.

Keywords: Collie eye anomaly, dogs, multiplex polymerase chain reaction assay, non-homologous end-joining factor 1 genotype.

Cervical anticancer activities of Annona squamosa Linn. leaf isolate

Research (Published online: 24-01-2022)

16. Cervical anticancer activities of Annona squamosa Linn. leaf isolate

Made Dira Swantara, Wiwik Susanah Rita, Made Asmarani Dira and Kadek Karang Agustina

Veterinary World, 15(1): 124-131

ABSTRACT

Background and Aim: Cancer is one of the leading causes of death, the need for new anticancer herbal drugs is becoming more urgent considering the side effects of synthetic drugs. This study aimed to determine the anticancer activity of isolates derived from the methanol extract of Annona squamosa Linn. leaves and to identify the compounds that have an active effect against HeLa cells.

Materials and Methods: The leaf metabolites of A. squamosa L. were extracted using methanol at room temperature (28°C) and were partitioned into n-hexane, chloroform, and n-butanol. The toxicity test of these extracts was conducted using a brine shrimp lethality assay. Furthermore, the most toxic extracts were separated and purified using silica gel column chromatography to yield four isolate fractions: FA, FB, FC, and FD. The most toxic isolates were tested for anticancer against HeLa cells, and their compounds were identified using liquid chromatography-mass spectrometry.

Results: The results showed that the most toxic isolate with an LC50 value of 100.00 ppm had a potency similar to that of an anticancer agent with an IC50 value of 70.9021 ppm. Furthermore, the five compounds identified in this isolate include (6S, 7aR)-6-hydroxy-4,4,7a-trimethyl-6,7-dihydro-5H-1-benzofuran-2-one or loliolide, cocamidopropyl betaine, N-[3- (dimethylamino)propyl]dodecanamide or lauramidopropyl dimethylamine, linolenic acid, and 1-dodecyl-2-azepanone or laurocapram.

Conclusion: It can be concluded that the leaf isolates of A. squamosa Linn. had shown anticancer activities against cervical cancer.

Keywords: Annona squamosa Linn, cervical anticancer activities, HeLa cells.

Comparison of three progesterone quantification methods using blood samples drawn from bitches during the periovulatory phase

Research (Published online: 24-01-2022)

15. Comparison of three progesterone quantification methods using blood samples drawn from bitches during the periovulatory phase

Hassan A. Hussein, Gerhard Schuler, Theresa Conze and Axel Wehrend

Veterinary World, 15(1): 119-123

ABSTRACT

Background and Aim: Measuring blood progesterone (P4) concentration has become an essential diagnostic tool in small animal reproductive medicine. Methods enabling precise and rapid on-site measurements are in high demand, especially for the optimization of breeding management in bitches. This study aimed to compare two commercial on-site methods (Speed™ P4, Virbac [M1] and mini VIDAS®, bioMérieux [M2]) and a well-established radioimmunoassay (RIA), which was used as a reference method.

Materials and Methods: Comparative measurements were performed on 52 blood serum samples collected from 45 clinically healthy bitches of different breeds. The dogs had been presented to determine the estrus cycle stage and predict the time of ovulation. Each sample was divided into three aliquots. In aliquot 1, P4 was measured immediately applying M2. Aliquots 2 and 3 were stored at –20°C until analysis was performed using RIA and M1. The consistency of the three methods was investigated by pairwise linear regression analyses.

Results: In RIA, the P4 concentrations ranged between 1.1 and 25.4 ng/mL. Regression analyses revealed highly significant (p<0.0001) positive correlations between the three methods applied (M1 vs. RIA: R=0.94; M2 vs. RIA: R=0.98; and M1 vs. M2: R=0.91).

Conclusion: The results show that the two commercial on-site methods tested exhibit approximately equal, high consistency with the radioimmunological reference method and can, therefore, be used beneficially in a clinical setting. However, biological interpretation of data must be performed in a method-specific manner.

Keywords: dog, enzyme-linked immunofluorescence, immunochromatography, progesterone, radioimmunoassay.

Seroprevalence of Toxoplasma gondii and associated alterations in hematology and serum biochemistry of one-humped camels (Camelus dromedarius) in Pakistan

Research (Published online: 22-01-2022)

14. Seroprevalence of Toxoplasma gondii and associated alterations in hematology and serum biochemistry of one-humped camels (Camelus dromedarius) in Pakistan

Aamir Shehzad, Awais Masud, Tabassam Fatima, Fraz Munir Khan, Saifur Rehman, Mustofa Helmi Effendi, Lucia Tri Suwanti, Iahtasham Khan, Wiwiek Tyasningsih, Shah Faisal, Zain Ul Abadeen and Samreen Bibi

Veterinary World, 15(1): 110-118

ABSTRACT

Background and Aim: Toxoplasma gondii is an intracellular protozoan that infects humans and animals. This study aimed to estimate the seroprevalence of T. gondii and the associated alterations in hematology and serum biochemistry of one-humped camels (Camelus dromedarius) in Mianwali district, Pakistan.

Materials and Methods: A total of 350 blood samples were obtained from male and female camels of different ages (≤3 years old, 4-6 years old, and ≥7 years old). To validate T. gondii antibodies, the collected samples were subjected to indirect enzyme-linked immunosorbent assay using purified recombinant micronemal protein 3 as an antibody catching antigen.

Results: The prevalence of T. gondii was 50.2% higher in male camels than in female camels (16.5%) (p<0.001). Furthermore, the prevalence of T. gondii in camels was directly proportional to age (p<0.001). It was 63.33% (57/90) in camels of ≥7 years of age, 32.54% in 4-6 years old age group, and 23.08% in ≤3 years old age group. The hematological analysis of infected camels revealed a significant increase in the values of glucocorticoid-remediable aldosteronism, lymphocyte percentage, monocyte percentage (MONO%), corpuscular hemoglobin (MCH), and procalcitonin. Furthermore, substantially higher levels of liver enzymes alanine aminotransferase, aspartate aminotransferase, and the macro-mineral potassium were found in the serum of T. gondii-infected camels.

Conclusion: The seropositivity of T. gondii is directly associated with the age and sex of camels, which may be considered as potential risk factors. Furthermore, T. gondii infection directly impacts the hemato-biochemistry of infected camels.

Keywords: biochemistry, camel, hematology, public health, seroprevalence, Toxoplasma gondii.

The enriched Y-bearing sperm combined with delayed fixed-time artificial insemination for obtaining male Simmental crossbred offspring

Research (Published online: 22-01-2022)

13. The enriched Y-bearing sperm combined with delayed fixed-time artificial insemination for obtaining male Simmental crossbred offspring

Dewa Ketut Meles, Imam Mustofa, Mas'ud Hariadi, Wurlina Wurlina, Suherni Susilowati, Anny Amaliya, Suparto Suparto and Rimayanti Rimayanti

Veterinary World, 15(1): 102-109

ABSTRACT

Background and Aim: The production of male calf beef cattle is an agricultural innovation needed to increase the farm's productivity as a provider of meat sources. This study aimed to determine the sex ratio of the offspring of cows inseminated with Y-bearing sperm enriched by Percoll density gradient centrifugation and swim up, combined with delayed fixed-time artificial insemination (FTAI).

Materials and Methods: Ejaculates of Simmental bulls were divided into four equal portions and grouped as T0 (control, non-sexed semen), T1 and T2 were sexed semen using Percoll density gradient centrifugation three and five levels, respectively, and T3 was sexed semen using swim-up. After the sex was sorted, the semen was diluted in a tris egg yolk extender, packaged in French mini-straws containing 50 million live sperm cells, and frozen. Pre-sexed, post-sexed, and post-thawed spermatozoa were evaluated based on progressive motility, viability, intact plasma membrane, and abnormality. The post-thawed semen of T0 was artificially inseminated to recipient cows at 12 h after onset of estrus (not delayed FTAI). Meanwhile, the delayed FTAI was conducted 18 20 h after onset of estrus using the T0, the best of T1 and T2, and the T3 post-thawed semen.

Results: The Percoll density gradient centrifugation reduced motility, viability, and intact plasma membrane but increased sperm abnormalities. Meanwhile, the swim up process increased motility, viability, and intact plasma membrane of sperm cells but decreased sperm abnormalities. Post-thawed semen decreased motility, viability, and intact plasma membrane of sperm cells but increased sperm abnormalities. The sex ratio of the Simmental crossbred offspring was 96.08% and 100% in T1 and T3, respectively, compared to 48.25% and 67.39% in T0 not delayed and delayed FTAI, respectively.

Conclusion: The Percoll density gradient centrifugation and swim-up methods are prospective for obtaining male offspring.

Keywords: agricultural innovation, farm productivity, motility, pregnancy rate, sperm morphologic abnormality, viability.

Assessment of aflatoxin M1 and B1 in some dairy products with referring to the analytical performances of enzyme-linked immunosorbent assay in comparison to high-performance liquid chromatography

Research (Published online: 22-01-2022)

12. Assessment of aflatoxin M1 and B1 in some dairy products with referring to the analytical performances of enzyme-linked immunosorbent assay in comparison to high-performance liquid chromatography

Raghda Mohamed Esam, Ragaa Shehata Hafez, Nagwa Ibrahim Mohamed Khafaga, Karima Mogahed Fahim and Lamiaa Ibrahim Ahmed

Veterinary World, 15(1): 91-101

ABSTRACT

Background and Aim: Aflatoxin M1 (AFM1) is a major fungal metabolite found in milk coming from aflatoxin B1 (AFB1) contaminated rations and is subsequently present in milk-based products demonstrating a serious public health hazard. This study aimed to investigate the levels of AFM1 and AFB1 in milk and some dairy products consumed widely by infants and children.

Materials and Methods: This study investigated the incidence of AFM1 in 105 samples of processed cheese, Ras cheese, and raw milk (35 of each) retailed in the Egyptian markets. The degree of sensitivity and accuracy was evaluated using the enzyme-linked immunosorbent assay (ELISA) method followed by the estimation of the positive samples using the high-performance liquid chromatography (HPLC) with fluorescence detection. Mold count was determined in the examined samples by investigating AFB1 content using HPLC.

Results: AFM1 was found in all investigated Ras cheese, raw milk, and 82.86% of the processed cheese samples with mean values of 51.05±6.19, 40.27±3.996, and 10.77±1.39 ng/kg, respectively. Moreover, there was statistically no significant difference between AFM1 levels in the core and crust parts of the tested Ras cheese. AFM1 contaminated Ras cheese and raw milk samples were 48.57% and 25.71%, which exceeded the European and Egyptian tolerance levels. Results showed an acceptable correlation between ELISA and HPLC methods with no significant difference (p>0.05). Alternatively, none of the examined samples proved to be contaminated with AFB1 despite the presence of mold with mean counts of 3.79±3.29, 4.39±4.34, and 4.84±4.29 log CFU/g in the examined processed cheese, Ras cheese, and raw milk samples, respectively.

Conclusion: Therefore, it is urgent to regularly inspect the contamination of animal feeds with AFB1 and apply special measures and novel techniques to protect the feed and food from public health hazards.

Keywords: aflatoxin B1, aflatoxin M1, enzyme-linked immunosorbent assay, high-performance liquid chromatography, mold, sensitivity.

Anthelmintic resistance status of goat gastrointestinal nematodes in Sing Buri Province, Thailand

Research (Published online: 20-01-2022)

11. Anthelmintic resistance status of goat gastrointestinal nematodes in Sing Buri Province, Thailand

Niorn Ratanapob, Nattanan Thuamsuwan and Suporn Thongyuan

Veterinary World, 15(1): 83-90

ABSTRACT

Background and Aim: Promotions of goat farming by both public and private sectors encouraged considerable goat raising in central Thailand. Gastrointestinal nematodes (GIN) infection is a major health and economic problem; however, evidence of resistance to broad-spectrum anthelmintics is frequently reported. Investigation of anthelmintic resistance (AR) status and identification of factors related to the development of AR is important components for sustainable GIN control. However, no information is available on this topic in the study area. The present study aimed to gather information on GIN control practices and to evaluate the effectiveness of albendazole, ivermectin, and levamisole for treating GIN infestation in goat herds in Sing Buri Province.

Materials and Methods: Twenty-nine herds were randomly selected. Information on management practices was collected by face-to-face interview using a structured questionnaire. Three field experiments for routinely used anthelmintics, including albendazole, ivermectin, and levamisole were conducted from June 2019 to November 2019. Fecal samples were collected pre- and post-treatment and examined for fecal egg count reduction to determine the status of anthelmintic resistance of goat GIN.

Results: Several improper practices were identified that lead to AR, especially chronic use of albendazole and ivermectin. All herds were considered resistant to albendazole and ivermectin, and levamisole resistant nematodes were detected in two herds. AR was strongly linked with the continuous use of anthelmintics.

Conclusion: Levamisole, which was still effective in the province, should be used with caution to minimize the selection of resistant strains. Farmers should be provided with updated information for sustainable parasite control. Further, the efficacy of anthelmintics should be routinely monitored.

Keywords: anthelmintic resistance, fecal egg count, gastrointestinal nematode, goat.

Effect of methanolic extract from Capsicum annuum against the multiplication of several Babesia species and Theileria equi on in vitro cultures, and Babesia microti in mice

Research (Published online: 20-01-2022)

10. Effect of methanolic extract from Capsicum annuum against the multiplication of several Babesia species and Theileria equi on in vitro cultures, and Babesia microti in mice

Mohamed Abdo Rizk, Shimaa Abd El-Salam El-Sayed, Mostafa Al-Araby and Ikuo Igarashi

Veterinary World, 15(1): 76-82

ABSTRACT

Background and Aim: Piroplasmosis is a serious disease that infects animals, inflicting significant economic losses in the livestock industry and animal trade worldwide. Anti-piroplasm drugs now on the market have demonstrated host toxicity and parasite resistance. As a result, developing more effective and safer anti-piroplasm drugs becomes an urgent issue. This study aimed to evaluate the inhibitory effect of Capsicum annuum methanolic extract (CA) against the growth of Babesia bovis, Babesia divergens, Babesia caballi, and Theileria equi in vitro and against B. microti in mice.

Materials and Methods: Fluorescence-based SYBR Green I assay was used to evaluate CA's inhibitory effect in vitro and in vivo when used either as a monotherapy or combined with diminazene aceturate (DA). The hematological parameters (HCT, hemoglobin, and red blood cells counts) were determined in the blood of mice every 96 h using Celltac α MEK-6450 electronic hematology analyzer.

Results: The in vitro growth of B. bovis, B. divergens, T. equi, and B. caballi was inhibited by CA in a dose-dependent manner with IC50 values of 4.87±1.23, 44.11±8.03, 8.23±2.54, and 1.26±0.50 mg/mL, respectively. In B. microti-infected mice, a combination therapy consisting of CA and a low dose of DA showed a significant (p<0.05) inhibition of B. microti growth nearly similar to those obtained by treatment with the full dose of DA.

Conclusion: The obtained results indicate that CA might be a promising medicinal plant for treating babesiosis, especially when used with a low dose of DA.

Keywords: Babesia, Capsicum annuum, combination therapy, in vitro, in vivo, Theileria.

Effect of gold nanoparticles shape and dose on immunological, hematological, inflammatory, and antioxidants parameters in male rabbit

Research (Published online: 19-01-2022)

9. Effect of gold nanoparticles shape and dose on immunological, hematological, inflammatory, and antioxidants parameters in male rabbit

Eman T. Mehanna, Basma S. A. Kamel, Dina M. Abo-Elmatty, Sameh M. Elnabtity, Manal B. Mahmoud, Mostafa M. Abdelhafeez and Ahmed Sabry S. Abdoon

Veterinary World, 15(1): 65-75

ABSTRACT

Background and Aim: Gold nanorods (AuNRs) have gained much attention in recent years due to their promising optical and chemical properties and are hence used in applied research and industrial nanotechnology. This study was designed to investigate the effect of gold nanoparticle shape (Gold nanorods vs. gold nanosphere) on immune response in rabbit.

Materials and Methods: Thirty New Zealand white rabbits were divided into six groups (n=5 rabbits). The first group is the control negative received an intravenous (IV) injection of normal saline 0.9%; the second group (vaccinated) is the control positive, and the other four groups were vaccinated and received a single-dose or repeated five consecutive IV doses of 300 μg/kg body weight 50 nm AuNRs or 50 nm gold nanosphere (50 nm AuNSs) dissolved in ultrapure water. Blood and serum were collected for the hematological and biochemical analysis.

Results: White blood cells (WBCs) count, lymphocytes, monocytes, eosinophils, and basophils showed significantly (p<0.05) higher values with the repeated-dose AuNRs. γ-globulin levels showed a significant difference after 15 days in the single-dose AuNSs. Single-dose AuNSs significantly (p<0.05) increased the immunoglobulin G (IgG) and significantly (p<0.05) decreased the tumor necrosis factor-alpha. In addition, it elicited a significant (p<0.05) decrease in the malondialdehyde levels and a significant (p<0.05) increase of the superoxide dismutase, glutathione peroxidase, and catalase levels. Moreover, evoked red blood cells count, mean corpuscular volume, and mean corpuscular hemoglobin were significantly (p<0.05) lower than the control group. The platelet count, lysozymes, and nitric oxide were significantly (p<0.05) higher in repeated-dose AuNRs.

Conclusion: The effect of AuNPs is shape and dose-dependent. The repeated 5 days IV 50 nm AuNRs doses over 15 days showed a significant antioxidant effect, with no considerable toxicity or vascular reactions.

Keywords: antioxidant's activity, dose, gold nanorods, gold nanospheres, hematological, immunological parameters, rabbit.

Emergence of extensively drug-resistant Aeromonas hydrophila complex isolated from wild Mugil cephalus (striped mullet) and Mediterranean seawater

Research (Published online: 18-01-2022)

8. Emergence of extensively drug-resistant Aeromonas hydrophila complex isolated from wild Mugil cephalus (striped mullet) and Mediterranean seawater

Hamza Mohamed Eid, Heba Sayed El-Mahallawy, Amany Mahmoud Shalaby, Hassnaa Mahmoud Elsheshtawy, Mera Mohamed Shetewy and Nada Hussein Eidaroos

Veterinary World, 15(1): 55-64

ABSTRACT

Background and Aim: Antibiotic resistance has been a progressively documented problem, resulting in treatment failure in humans and animals. This study aimed to investigate the antimicrobial susceptibility and virulence of extensively drug-resistant (XDR) Aeromonas spp. in wild Mugil cephalus and its surrounding seawater along the coastal road of Port Said, Egypt.

Materials and Methods: Specimens were examined bacteriologically, confirmed biochemically, and tested for their sensitivity against 11 antimicrobial agents. Molecular confirmation of the obtained isolates by 16S rRNA was performed, followed by the detection of antimicrobial resistance and virulence genes.

Results: Aeromonas spp. was recovered from fish (44%) and water samples (36%). A. hydrophila was the most prevalent identified strain, followed by Aeromonas sobria, Aeromonas caviae, and Aeromonas schubertii. Moreover, 90% of the tested isolates were multidrug-resistant (MDR), while 26.67% were XDR. Tested isolates were resistant to β-lactams and sulfonamides (100%), oxytetracycline (90%), and streptomycin (62.22%) but completely susceptible to cefotaxime. XDR isolates successfully amplified resistance genes (blaTEM, sul1, and tetA(A)) but not the (aadA1) gene, although there was phenotypic resistance to streptomycin on plates. All XDR isolates carry the cytotoxic enterotoxin gene (act), but alt gene was detected in only one isolate (12.5%).

Conclusion: Data in this study provide a recent update and highlight the role of wild mullet and seawater as reservoirs for MDR and XDR Aeromonas spp. that may pose a risk to humans as food-borne infection or following direct contact.

Keywords: Aeromonas hydrophila complex, antimicrobial resistance, Mugil cephalus, resistance genes, Seawater, virulence genes.

Glutathione-S-transferase activity in various organs of Crocodylus siamensis and its attenuation role in aflatoxin B1-induced cell apoptosis in human hepatocarcinoma cells

Research (Published online: 18-01-2022)

7. Glutathione-S-transferase activity in various organs of Crocodylus siamensis and its attenuation role in aflatoxin B1-induced cell apoptosis in human hepatocarcinoma cells

Piriyaporn Thiendedsakul, Pitchaya Santativongchai, Prapassorn Boonsoongnern, Rungrueang Yodsheewan and Phitsanu Tulayakul

Veterinary World, 15(1): 46-54

ABSTRACT

Background and Aim: The crocodile is a model for studying relevant sources of environmental contamination. They were determined an appropriate biomonitoring species for various toxins. The cytosolic and microsomal fraction of crocodiles plays a role in detoxifying xenobiotics. Cytochrome P450 1A2 (CYP1A2) metabolizes aflatoxin B1 (AFB1) to aflatoxin M1, while glutathione-S-transferase (GST) catalyzes carcinogenic agents. This study aimed to investigate the GST activity in various organs of Crocodylus siamensis. Further, the fate of microsomal and cytosolic fractions from various crocodile organs against AFB1-induced apoptosis in human hepatocarcinoma (HepG2) cells was investigated.

Materials and Methods: The liver, lungs, intestines, and kidneys tissues from a 3-year-old crocodile (C. siamensis) (n=5) were collected. The cytosolic and microsomal fraction of all tissues was extracted, and protein concentrations were measured with a spectrophotometer. Subsequently, a comparison of GST activity from various organs was carried out by spectrophotometry, and the protective effects of CYP450 and GST activity from various crocodile organs were studied. In vitro AFB1-induced apoptosis in HepG2 cells was detected by reverse transcription-quantitative polymerase chain reaction. Comparisons between the metabolisms of the detoxification enzyme in organs were tested using the Kruskal–Wallis one-way analysis of variance and Dunn's multiple comparison tests. All kinetic parameters were analyzed using GraphPad Prism software version 5.01 (GraphPad Software Inc., San Diego, USA).

Results: Total GST activity in the liver was significantly higher than in the kidneys, intestines, and lungs (p<0.05, respectively). The highest GST pi (GSTP) activity was found in the liver, while the highest GST alpha-isoform activity was in the crocodile lung. The kinetics of total GST and GST mu activity in the liver had the highest velocity compared to other organs. In contrast, the kinetics of GSTP enzyme activity was the highest in the intestine. The in vitro study of microsome and cytosol extract against apoptosis induced by AFB1 revealed that the level of messenger RNA expression of the Bax and Bad genes of HepG2 cells decreased in the treatment group in a combination of cytosolic and microsomal fractions of the crocodile liver but not for Bcl-2. Interestingly, the downregulated expression of Bax and Bad genes was also found in the microsome and cytosol of crocodile kidneys.

Conclusion: The crocodile liver revealed very effective GST activity and expression of the highest kinetic velocity compared to other organs. The combination of liver microsomal and cytosolic fractions could be used to prevent cell apoptosis induced by AFB1. However, further study of the molecular approaches to enzyme activity and apoptosis prevention mechanisms should be carried out.

Keywords: aflatoxin B1, apoptosis, Crocodylus siamensis, glutathione-S-transferase, human hepatocarcinoma cells.

Effect of barley straw and Egyptian clover hay on the rumen fermentation and structure and fibrolytic activities of rumen bacteria in dromedary camel

Research (Published online: 17-01-2022)

6. Effect of barley straw and Egyptian clover hay on the rumen fermentation and structure and fibrolytic activities of rumen bacteria in dromedary camel

Alaa Emara Rabee

Veterinary World, 15(1): 35-45

Background and Aim: Understanding the regulations of rumen microbiota and their fibrolytic capabilities under different forages are essential to improve rumen fermentation and animal feed efficiency. This study aimed to evaluate the changes in the rumen fermentation and the structure and fibrolytic activities of rumen bacteria in camels fed barley straw and Egyptian clover hay.

Materials and Methods: Three fistulated camels were fed a diet containing barley straw for 30 days; then transitioned to a diet containing Egyptian clover hay for 30 days. In addition, bacterial media enriched with xylan and different cellulose sources, namely, filter paper, wheat straw, and alfalfa hay, were used to evaluate the ability of camel rumen bacteria to produce xylanase and cellulase enzymes.

Results: The camel group fed Egyptian clover hay showed higher crude protein intake, rumen ammonia, total volatile fatty acids, and acetic acid. Moreover, the camel group fed barley straw showed higher neutral detergent fiber intake, rumen pH, and propionic and butyric acids. Principal component analysis showed that bacterial communities were separated based on the forage type. Forage type affected the composition of rumen bacteria and most of the bacterial community was assigned to phylum Bacteroidetes and Firmicutes. Egyptian clover hay diet increased the proportions of genus Prevotella and Ruminococcus; while fed barley straw diet increased the Butyrivibrio, RC9_gut_group, and Fibrobacteres. The bacterial culture of the Egyptian clover hay fed group produced the greatest xylanase and the bacterial culture of the barley straw fed group produced the maximum cellulase.

Conclusion: Egyptian clover hay is recommended to feed camels in intensive production. Moreover, the bacterial community in the camel rumen is a promising source of lignocellulolytic enzymes.

Keywords: Camelus dromedaries, cellulase and xylanase, forage type, hay and straw, rumen bacteria.

Seroprevalence and risk factors of porcine cysticercosis: A cross-sectional study in Indonesia

Research (Published online: 17-01-2022)

5. Seroprevalence and risk factors of porcine cysticercosis: A cross-sectional study in Indonesia

Annytha Detha, Putri Pandarangga and Yunita Nope

Veterinary World, 15(1): 30-34

ABSTRACT

Background and Aim: Cysticercosis is a zoonotic disease with a global concern. Estimation of the prevalence and identification of potential risk factors are necessary for the prevention and control of the disease. This study aimed to estimate the seroprevalence of cysticercosis and the correlation of the increased prevalence with several potential risk factors.

Materials and Methods: The seroprevalence of cysticercosis was conducted using an enzyme-linked immunosorbent assay (ELISA), developed by the Institute of Tropical Medicine Antwerp, to detect Cysticercus cellulosae. This study used serum samples from 62 pigs taken from two regencies on Timor Island. The data analysis was performed using SPSS software 20.0 (IBM Corp., NY, USA) to evaluate ELISA results and the strength of the relationship between risk factors and the prevalence of disease using the odds ratio (OR).

Results: Serum samples from 18 out of the 62 pigs were found to be positive; the seroprevalence of cysticercosis was 29%. The results showed that an extensive farming system led to a higher prevalence of cysticercosis compared to an intensive farming system, namely, 10 out of 18 (56.6%), and that the possibility of identifying cysticercosis in pigs in an extensive farming system was 5 times greater than that in pigs in an intensive farming system. In addition, the results showed that nine out of 18 households who did not have toilet facilities were found to be seropositive, indicating a significant relationship between the risk factor of toilet availability with cysticercosis in pigs, with an OR of 4.5. In addition, the results showed that there was no significant relationship between the risk factor of the feed source and the prevalence of cysticercosis in pigs.

Conclusion: It can be concluded that the seroprevalence of cysticercosis was 29% in domestic pigs of Timor Island. The risk factors of an extensive pig farming system and toilet availability in community houses were significantly related to the possibility of cysticercosis on Timor Island.

Keywords: cysticercosis, epidemiology, risk factor, seroprevalence.