First report on molecular characteristics and risk factor analysis of Ehrlichia canis in dogs in Khon Kaen, Thailand

Research (Published online: 31-01-2022)

29. First report on molecular characteristics and risk factor analysis of Ehrlichia canis in dogs in Khon Kaen, Thailand

Thongphet Mitpasa, Biethee Rani Sarker, Arayaporn Macotpet, Pattara-Anong Bupata, Somboon Sangmaneedet and Weerapol Taweenan

Veterinary World, 15(1): 232-238

ABSTRACT

Background and Aim: Ehrlichia canis is a well-known cause of both anemia and thrombocytopenia in dogs. There are insufficient epidemiological data on this blood parasite in Thailand and the association of infections with hematological abnormalities. This study aimed to analyze the molecular characteristics and to identify E. canis as well as the risk factors associated with E. canis infection in dogs in Khon Kaen, Thailand.

Materials and Methods: Blood samples from 126 dogs that visited animal clinics were subjected to molecular detection using nested polymerase chain reaction for E. canis 16S rRNA gene. The risk factors and hematological profiles associated with the infection were analyzed using the logistic regression test in program SPSS version 19.

Results: Forty-one dogs were infected, indicating a 32.5% molecular infection rate of E. canis. The factors significantly associated with E. canis infection include animal housing status, low packed cell volume, low red blood cell count, and low platelets (p<0.05). Ten positive samples were amplified, sequenced, and phylogenetically analyzed. Sequence and phylogenetic analysis confirmed the current ten samples as E. canis compared with reference sequences in GenBank, using the BLAST program hosted by NCBI, which showed 99.74-100% similarity.

Conclusion: This study provided the first data of infection rate of E. canis using nested PCR and molecular characteristics of E. canis in randomly selected domestic dogs in Khon Kaen, Thailand.

Keywords: Ehrlichia canis, molecular characteristics, nested polymerase chain reaction, phylogenetic analysis, Thailand.

Prospects of bacteriophage collections in disinfectant applications

Research (Published online: 31-01-2022)

28. Prospects of bacteriophage collections in disinfectant applications

Samat Serikovich Issabekov, Nazym Syrymkyzy Syrym, Aidar Adilkhanovich Sambetbayev, Kuantar Daulenovich Alikhanov and Bolat Amanbaevich Yespembetov

Veterinary World, 15(1): 220-231

ABSTRACT

Background and Aim: The search and development of disinfectants is promising worldwide. However, there are currently no international regulations governing the testing and registration of germicidal agents. Moreover, the number of safety requirements for disinfectants for human, animal, and environmental health has increased. This research aimed to evaluate the prospects of using a collection of bacteriophages for disinfectant purposes.

Materials and Methods: The objects of research were bacteriophages isolated from a total of 129 environmental samples obtained from seven sources in and around livestock buildings: (1) Feed residues from feeders and automatic drinkers; (2) washouts from floors, walls, and posts; (3) soil from underneath floors; (4) bedding; (5) sewage; (6) ponds; and (7) soil from paddocks. The corresponding strains were used as indicator test cultures for bacteriophages. The authors employed the following methods to work with bacteriophages: (a) Bacteriophage isolation methods, (b) the Appelman method (i.e., serial dilutions), (c) the Grazia method (i.e., agar layers), (d) phage titration on solid media, and (e) the bacterial phagotyping method.

Results: The results of the analysis on the bacteria of the Enterobacteriaceae family isolated 11 bacteriophages; one bacteriophage is specific to Pseudomonas aeruginosa, and another one is specific to Brucella abortus. The results also indicate that all bacteriophage strains of the Enterobacteriaceae family demonstrate lysis at a pH of 7.0. In addition, this polyphage lyses all strains of sensitive bacterial cultures. The optimum temperature for the cultivation of bacteriophages is 35°C. While using electron microscopy to study the consortium of bacteriophages, clearly distinguishable virions of bacteriophages were found in the microscope field of view.

Conclusion: The main parameters for the production of polyphages include the ratio of the bacteriophage and its corresponding bacteriophage-sensitive culture, the pH of the cultivation medium, and the cultivation time of the bacteriophage system as well as the sensitive bacterium. With regard to the aforementioned parameters, the results indicate that the average value for all bacteriophages is 1:2, and the average cultivation medium pH is 7.0 for all bacteriophages. The average cultivation time for all bacteriophages is 18-24 h.

Keywords: bacteriophages, biological preparation, disinfection, lytic activity, strain, titer.

Epizootiological aspects of natural nidality of Ixodes tick-borne borreliosis in the Moscow region (Russian Federation)

Research (Published online: 30-01-2022)

27. Epizootiological aspects of natural nidality of Ixodes tick-borne borreliosis in the Moscow region (Russian Federation)

Almas Mukhametov, Mikhail Osadchuk, Iza Berechikidze and Nikolay Pronkin

Veterinary World, 15(1): 213-219

ABSTRACT

Background and Aim: At present, tick-borne borreliosis is the most common infectious disease transmitted by ticks in Europe, Asia, and North America. This study aimed to examine the epizootiological aspects of the natural nidality of tick-borne borreliosis in Moscow region (the Russian Federation).

Materials and Methods: A total of 2,537 ticks representing two species were collected, namely, Ixodes ricinus and Dermacentor reticulatus. The activity, number of ticks, and Borrelia infestation rates were investigated during a high season, that is, from early spring to mid-autumn.

Results: In May, amount of I. ricinus spp. was found 2.5 times more than those representing D. reticulatus spp. (p≤0.01). In June, August, and September, the amount of I. ricinus was 9.0 (p≤0.0001), 2.0 (p≤0.05), and 5.0 times higher, respectively, compared to D. reticulatus. In the first 10 days of April, the amount of D. reticulatus was 3 times higher than that of I. ricinus (p≤0.02); in the next 10 days, their amounts were equal (p≥0.05) and in the last 10 days the amount of I. ricinus exceeded that of D. reticulatus (p≤0.05) by 1.5 times. In general, Borrelia afzelii, and Borrelia garinii, were detected. In addition, the naturally occurring tick-borne borreliosis pesthole was revealed in the Moscow region.

Conclusion: Borrelia infection rates for ticks comprise 30%. An increase in Borrelia tick infestation was detected within the vicinity of populated areas. The amount of ticks directly depends on the temperature (20°C-25°C) and moisture (from 50%) values.

Keywords: borreliosis, Dermacentor reticulatus, infection, Ixodes ricinus, Ixodes ticks.

Molecular characterization of genes responsible for biofilm formation in Staphylococcus aureus isolated from mastitic cows

Research (Published online: 30-01-2022)

26. Molecular characterization of genes responsible for biofilm formation in Staphylococcus aureus isolated from mastitic cows

Eman Shafeek Ibrahim, Amany Ahmed Arafa, Sohad Mohamed Dorgam, Rasha Hamdy Eid, Nagwa Sayed Atta, Wahid Hussein El-Dabae and Eslam Gaber Sadek

Veterinary World, 15(1): 205-212

ABSTRACT

Background and Aim: Mastitis is considered a significant disease of lactating animals. There are new attitudes for recognizing genes responsible for causing this disease to overcome and change the manipulation of this problem. This study aimed to isolate and identify Staphylococcus aureus strains from mastitic bovine animals and detect some specific biofilm-forming genes (icaA, icaD, and biofilm-associated protein [bap] genes clfA, fnbA, agrI, agrII, agrIII, agrIV, and cna).

Materials and Methods: A total of 121 mastitic milk samples were analyzed using biochemical tests (catalase test, oxidative-fermentative test, and coagulase test) and Gram stain. Multiplex polymerase chain reaction was applied to characterize biofilm genes (icaA, icaD, bap, clfA, and fnbA) in addition to (agrI, agrII, agrIII, agrIV, and cna).

Results: Among the 121 milk samples, 35 staphylococci isolates were derived with an incidence of 28.92% (35/121); among them, 19 are coagulase positive. Ninety percent of the isolates had ica genes (icaA and icaD) while bap gene was not recognized in any isolate. In addition, the incidence of fnbA, can, and clfA was 89.5% each. The prevalence of agr specific groups (agrI, agrII, agrIII, and agrIV) was 78.9%, 52.6%, 10.5%, and 15.8%, respectively.

Conclusion: This study concluded that S. aureus has variant mechanisms of pathogenicity to form biofilm devoid of carrying a specific gene.

Keywords: biofilm genes, mastitis, molecular identification, Staphylococcus aureus.

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.

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.