Molecular prevalence of Dirofilaria immitis and Wolbachia infections in pet and semi-domesticated cats in Bangkok, Thailand

Research (Published online: 03-02-2022)

1. Molecular prevalence of Dirofilaria immitis and Wolbachia infections in pet and semi-domesticated cats in Bangkok, Thailand

Naris Thengchaisri, Tawin Inpankaew, Surapong Arthitwong, Jörg M. Steiner and Panpicha Sattasathuchana

Veterinary World, 15(2): 239-243

ABSTRACT

Background and Aim: Although cats are not natural hosts for heartworm infections (Dirofilaria immitis), evidence suggests that feline heartworm disease can be detrimental because of a severe inflammatory response. Recent studies have found that infection with bacteria of the genus Wolbachia is the principal cause of acute inflammatory filaria disease; nonetheless, the prevalence of cats naturally infected with heartworms and Wolbachia remains unclear. This study aimed to estimate the prevalence and current distribution of feline heartworm disease and its association with Wolbachia infection in pet and semi-domesticated cats in Bangkok, Thailand.

Materials and Methods: A total of 260 cats (130 pet cats and 130 semi-domesticated cats) were enrolled in this study. Blood samples were placed into ethylenediaminetetraacetic acid tubes for hematological analysis and DNA extraction. A polymerase chain reaction (PCR) was performed to analyze samples for the presence of D. immitis and Wolbachia infections.

Results: The prevalence (95% confidence interval [CI]) of D. immitis infection in pet, semi-domesticated, and all cats were 3.9% (1.3-8.8%), 27.7% (20.2-36.2%), and 19.6% (15.0-25.0%), respectively. The prevalence (95% CI) of Wolbachia infection in pet, semi-domesticated, and all cats were 18.5% (12.2-26.2%), 31.5% (23.7-40.3%), and 25.0% (19.9-30.7%), respectively. The prevalence of D. immitis and Wolbachia infections in semi-domesticated cats was significantly higher than in pet cats (p=0.002 and p=0.022, respectively). There was a significant association between D. immitis and Wolbachia infections (p<0.001). There was also a significant association between D. immitis infection and the presence of eosinophilia (p<0.045).

Conclusion: From the PCR analysis, it can be concluded that semi-domesticated cats were at higher risk for D. immitis infection than pet cats. There was a significant association between positive D. immitis infection and positive Wolbachia infection. Combinations of anthelmintic and antimicrobial therapy should be considered in heartworm-positive cats.

Keywords: cat, heartworm, prevalence, Wolbachia.

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.