Viability of Toxoplasma gondii tachyzoites in different conditions for parasite transportation

Research (Published online: 29-01-2022)

25. Viability of Toxoplasma gondii tachyzoites in different conditions for parasite transportation

Thi Thuy Nguyen, Ketsarin Kamyingkird, Waraphon Phimpraphai and Tawin Inpankaew

Veterinary World, 15(1): 198-204

ABSTRACT

Background and Aim: Toxoplasma gondii tachyzoite is the infective stage that causes acute infection, leading to severe toxoplasmosis. The tachyzoite stage has been extensively used for several inoculation purposes, including antigen production, immunological studies, nutrition mechanisms, and in vitro drug trials. The use of fresh tachyzoites is required for inoculation in either in vitro or in vivo studies. However, there is a lack of information on preserving live tachyzoites during transportation from laboratories to inoculation sites. Therefore, this study aimed to validate suitable preservative conditions for maintaining live parasites by determining the survival and viability of T. gondii tachyzoites on the basis of different media, temperatures, and incubation times.

Materials and Methods: The free live T. gondii tachyzoites were evaluated on their viability when maintained in different media without 5% Carbon dioxide (CO2). The purified tachyzoites of the RH and PLK strains were individually suspended in normal saline (NS), phosphate-buffered saline (PBS), minimum essential medium (MEM), and MEM with 10% fetal bovine serum (MEM-FBS) and incubated for 6 h at ice-cold (IC; 3-9°C) and room temperature (RT; 25°C). Parasite survival was measured at the 0, 1st, 2nd, 3rd, 4th, 5th, and 6th h post-incubation using the trypan blue exclusion test.

Results: The viability was in the range of 85.0%–91.0% for IC using NS and 81.0%–85.1% (IC) and 75.3%–77.5% (RT) using PBS. The viability was approximately 75.0%–83.0% (IC) and 70.0%–79.0% (RT) using MEM and MEM-FBS. There was a significant difference in the viability between the seven periods on the basis of one-way repeated Analysis of variance and Friedman analyses. Parasite survival slightly reduced (20.0%–30.0%) in NS and MEM-FBS at both temperatures during incubation. Notably, PBS could not support tachyzoite viability after 3 h post-incubation.

Conclusion: NS was a suitable preservative for maintaining purified T. gondii tachyzoites during transportation at IC and RT without 5% CO2 supplementation. This could be a valuable medium for parasite transportation, especially when there is a large distance between the laboratory and inoculation site.

Keywords: preservation time, T. gondii tachyzoites, temperature, transportation, viability.

Treatment efficacy of Thunbergia laurifolia, Curcuma longa, Garcinia mangostana, and Andrographis paniculata extracts in Staphylococcus aureus-induced rabbit dermatitis model

Research (Published online: 29-01-2022)

24. Treatment efficacy of Thunbergia laurifolia, Curcuma longa, Garcinia mangostana, and Andrographis paniculata extracts in Staphylococcus aureus-induced rabbit dermatitis model

Charinya So-In and Nuchsupha Sunthamala

Veterinary World, 15(1): 188-197

ABSTRACT

Background and Aim: Dermatitis is a soft-tissue infection caused by Staphylococcus aureus. The recurrence of inflammatory skin is linked to clinical manifestations. Anti-inflammatory cytokines, which are essential for tissue damage, are released by bacteria through skin tissues. Oxidative stress causes inflammatory cells to necrotize and reduces their antioxidant profile, resulting in toxic damage to surrounding tissues. Although studies on the antibacterial effects of Thunbergia laurifolia Lindl., Curcuma longa L., Garcinia mangostana L., and Andrographis paniculata (Burm.). Bacterial infection of S. aureus have been conducted, most of these studies have been in vitro and were not related to the rabbit model. In addition, anti-inflammatory and antioxidant studies need to be evaluated. Thus, this study aims to compare the antibacterial, anti-inflammatory, and antioxidant properties of four local herbs with a standard antibiotic in S. aureus-induced rabbit dermatitis model.

Materials and Methods: The skin of New Zealand white rabbits were artificially wounded using a sterile blade and then infected with S. aureus. The rabbits were divided into seven groups, each with three rabbits (Total 21 rabbits): The first group was the no infection group (no infection and no treatment with scarification), the second group was the no treatment group (S. aureus infection of the wound but no treatment), and the other five treated groups were T. laurifolia, C. longa, G. mangostana, A. paniculata, and bacitracin cream, all of which involved wound infection and treatments. The treatment lasted for 7 days. The antibacterial, anti-inflammatory, and antioxidant properties after treatment were measured.

Results: The efficacy of T. laurifolia, C. longa, G. mangostana, and A. paniculata was similar to that of an antioxidant and free radical scavenging property. The bacterial infection process gradually reduced the activities of antioxidant systems (i.e., enzymatic levels and gene expressions) and total glutathione. However, the activities of the antioxidant system were steadily increased when treated with herbal extracts. During bacterial invasion of the skin, the concentration of thiobarbituric acid reactive molecules, the level of lipid peroxidation, and the expression of anti-inflammatory cytokine genes were increased. All these were decreased when herbal extracts were used to treat the lesion.

Conclusion: It can be concluded that T. laurifolia, C. longa, G. mangostana, and A. paniculata extract have antibacterial, anti-inflammatory, and antioxidant properties and are effective antibacterial agents. G. mangostana is the most effective herbal extract for antidermatitis and has the potential to be used as an alternative topical treatment.

Keywords: antibacterial, antidermatitis, anti-inflammatory, antioxidant, Garcinia mangostana, lipid peroxidation.

The improvement of insulin level after hydrogen-rich water therapy in streptozotocin-induced diabetic rats

Research (Published online: 28-01-2022)

23. The improvement of insulin level after hydrogen-rich water therapy in streptozotocin-induced diabetic rats

Ekowati Retnaningtyas, Budi Susatia, Siti Nur Arifah and Sri Rahayu Lestari

Veterinary World, 15(1): 182-187

ABSTRACT

Background and Aim: Water plays a pivotal role in the body. Alteration of the fluid balance promotes metabolic disorder, thus leading to the development of various diseases, such as diabetes mellitus (DM). Hydrogen-rich water (HW) is recognized as a novel antioxidant. This study aimed to investigate the role of HW on insulin, insulin receptor (IRs), and superoxide dismutase (SOD) levels in streptozotocin (STZ)-induced diabetic rats.

Materials and Methods: A total of 30 male Wistar rats were randomly divided into five groups: Normal (N), DM rats, DM+metformin (DM+Met, 45 mg/kg body weight [BW]), DM+Met+HW, and DM+HW. DM rats were induced by feeding them a high-fat diet for 30 days and then injecting with repeated low doses of STZ (35 mg/kg BW) intraperitoneally. Fresh HW was administered orally and ad libitum for 14 days. Insulin, IRs, and SOD were observed in each group.

Results: HW therapy increased the level and expression of insulin and IRs. In addition, treatment with HW also elevated the SOD levels in the serum and liver. The study results indicated no significant differences between the administration of HW and metformin.

Conclusion: HW has antioxidant activity in STZ-induced DM rats, increasing insulin, IRs, and SOD.

Keywords: diabetes mellitus, hydrogen-rich water, insulin receptor, insulin, superoxide dismutase.

Synbiotic goat milk kefir improves health status in rats fed a high-fat and high-fructose diet

Research (Published online: 28-01-2022)

22. Synbiotic goat milk kefir improves health status in rats fed a high-fat and high-fructose diet

Nurliyani Nurliyani, Eni Harmayani and Sunarti Sunarti

Veterinary World, 15(1): 173-181

ABSTRACT

Background and Aim: Kefir, a natural probiotic containing bacteria and yeast, is a fermented milk product, whereas glucomannan from porang tuber (Amorphophallus oncophyllus) is prebiotic in vivo. Simvastatin is a potent lipid-lowering statin that can be utilized for pharmacological therapy in obesity. This study aimed to determine the effect of goat milk kefir supplemented with porang glucomannan (synbiotic kefir) and goat milk kefir without glucomannan (probiotic kefir) on blood glucose, hemoglobin A1c (HbA1c), free fatty acids (FFAs), tumor necrosis factor-alpha (TNF-α), gene expression of peroxisome proliferator-activated receptor gamma (PPARγ), and insulin-producing cells in rats fed a high-fat and high-fructose (HFHF) diet.

Materials and Methods: Male Sprague-Dawley rats were divided into five dietary groups: (1) Normal control, (2) rats fed HFHF, (3) rats fed HFHF+probiotic kefir, (4) rats fed HFHF+synbiotic kefir, and (5) rats fed HFHF+simvastatin. All of these treatments were administered for 4 weeks.

Results: There were no significant differences in plasma glucose levels in HFHF diet-fed rats before and after treatment. However, plasma HbA1c and TNF-α decreased, and FFAs were inhibited in rats after treatment with synbiotic kefir. Synbiotic kefir decreased the gene expression of PPARγ2 in HFHF diet-fed rats but did not affect the total number of islets of Langerhans and insulin-producing cells.

Conclusion: Synbiotic kefir improved the health of rats fed an HFHF diet by decreasing HbA1c, TNF-α, and PPARγ2 gene expression and preventing an increase in FFAs.

Keywords: health status, high fat-high-fructose diet, porang glucomannan, simvastatin, synbiotic kefir.

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.