First report of acanthocephalan parasite in wild-caught Asian vine snake (Ahaetulla prasina) in Indonesia

Research (Published online: 17-02-2023)

10. First report of acanthocephalan parasite in wild-caught Asian vine snake (Ahaetulla prasina) in Indonesia

Aditya Yudhana, Ratih Novita Praja, and Ryanka Edila

Veterinary World, 16(2): 317-321

ABSTRACT

Background and Aim: Exotic pet snakes are more susceptible to infection, especially parasitic helminths than wild-caught. There is no comprehensive report on the prevalence of acanthocephalan parasite infection in Indonesian snakes. Therefore, this study aimed to estimate the prevalence rate and to identify the acanthocephalan infection in wild-caught Asian vine snake (Ahaetulla prasina) from the Mojokerto District, East Java, Indonesia.

Materials and Methods: A total of 60 snakes were collected from the local sellers in the Mojokerto District, East Java, Indonesia. Then, snakes were euthanized and necropsied to observe various predilections of acanthocephalan larval stage (cystacanth). Morphological identification of the cystacanth was conducted using the carmine staining method and microscopic examination.

Results: Acanthocephalan infection was recorded with a high prevalence rate of 80.06%. A total of 696 cystacanths were examined from the muscle, subcutaneous tissues, and visceral with 32.90, 16.37, and 50.71% intensity rates, respectively.

Conclusion: Acanthocephalan prevalence rate was recorded at 80.06% in this study. Constant disease monitoring is necessary, considering wild-caught Asian vine snakes were susceptible host and lack of data regarding parasitological surveys. Therefore, further studies are needed in new areas and various species of wild-caught snakes in Indonesia, because of the potential of parasitic helminth transmission between snake and other reptiles.

Keywords: Acanthocephalan, Ahaetulla prasina, infectious disease, neglected disease.

The relationship between ovarian hormones and mast cell distribution in the ovaries of dromedary camel (Camelus dromedaries) during the follicular wave

Research (Published online: 17-02-2023)

9. The relationship between ovarian hormones and mast cell distribution in the ovaries of dromedary camel (Camelus dromedaries) during the follicular wave

Ragab H. Mohamed, Nasra A. Yousef, Mahmoud Awad, Rasha S. Mohamed, Fatma Ali, Hassan A. Hussein, and Axel Wehrend

Veterinary World, 16(2): 309-316

ABSTRACT

Background and Aim: Mast cells (MCs) play an essential role in regulating tissue homeostasis through various non-allergic immune reactions. This study aimed to describe the salient features of MCs during different phases of the estrous cycle and evaluate the relationship between ovarian hormones and the presence of MCs in camel ovaries.

Materials and Methods: Genital tracts (n = 28) of healthy, non-pregnant camels were collected from a local slaughterhouse. The follicular wave stage was determined according to structures on the ovaries using an ultrasound device. Stages were classified as "growing" (n = 12, FØ = 0.3–0.8 cm), "mature" (n = 9, FØ = 0.9–2.2 cm), or "regression" phase (n = 7, FØ >2.5). Blood samples were collected at slaughter to determine serum estradiol-17β and progesterone levels using an immunoassay. Safranin-O, periodic acid/Schiff, alcian blue, or methylene blue stains were used to detect MCs.

Results: Follicular numbers at the growing, mature, and regression phases were determined to be 36, 14, and 7 follicles, respectively. Mast cells were widely but sparsely distributed within the ovarian tissue (9.3 MCs in the growing phase, 10.7 in the mature phase, and 7.0 in the regression phase). Typical histological features of MCs were observed in ovarian stromal tissue. Some MCs were found in the interstitial tissue, either near the follicular wall or the interstitial gland. Mast cells were present at a higher density during the mature phase than in the growing and regression phases in the ovarian matrix. A significantly reduced presence of MCs was found in the regression phase than in both the growing and mature phases (p < 0.05). A very strong positive correlation was observed between serum estradiol-17β concentrations and MC density in the ovaries (r = 0.9; p < 0.001). In addition, a strong negative correlation (r = –0.65; p = 0.03) was observed between the presence of MCs and serum progesterone concentrations.

Conclusion: These findings suggest that the follicular wave phase and the associated hormonal concentration induce changes in the number of MCs in the camel ovary.

Keywords: camel, follicular wave, immunohistochemistry, mast cell, ovarian hormones.

Therapeutic approaches for anti-sperm-antibodies in the testicular sperm aspiration rat model

Research (Published online: 15-02-2023)

8. Therapeutic approaches for anti-sperm-antibodies in the testicular sperm aspiration rat model

Abdel-Kader A. Zaki, Fahad S. Aldahmashi, Abd El-Nasser A. Madboli, Kamal A. Attia, Fahad S. Almulhim, and Saleh M. Albarrak

Veterinary World, 16(2): 296-308

ABSTRACT

Background and Aim: Anti-sperm antibodies (ASAs) treatment continued to be neglected. This study aimed to generate ASAs using the testicular sperm aspiration (TSA) rat model, which allowed for investigation of four distinct therapeutic approaches to find potential treatments for ASAs.

Materials and Methods: Adult Wistar albino male rats were divided into six equal groups (n = 12). The negative control group underwent scrotal sac surgery without having their testicles punctured. Punctures were made in the remaining 5 groups, with one group left untreated to serve as the positive control group. The remaining 4 groups were treated with either dexamethasone (DEX), azathioprine (AZA), frankincense, or anti-ASAs secondary antibodies. For 10 weeks, serum samples were collected every 2 weeks for specific quantification of ASAs. Testis and epididymis tissues were collected for histopathological analysis.

Results: The ASAs concentrations of the positive controls were significantly higher (p ≤ 0.001) than their negative control counterparts during the examined weeks. However, The ASAs indices (%) differed according to the treatment type. While the ASAs indices at the 2nd and 4th weeks in the AZA-treated group were significantly reduced compared to the positive control group (p ≤ 0.001), no significant differences were observed at any of the sample collection week for the DEX-treated rats. The ASAs indices were significantly decreased only at weeks 6 and 8 of treatment in the frankincense-treated group (p ≤ 0.001). In the secondary antibodies-treated group, the antibody indices were significantly decreased in all weeks except for samples collected at week 4 (p ≤ 0.001). The testosterone levels reverted to normal only in TSA rats treated with either Frankincense or secondary antibodies, as they were significantly higher than the positive controls (p ≤ 0.05). Tissue samples from the secondary antibody-treated rats showed a generally normal histological appearance.

Conclusion: This study tried to offer realistic therapy suggestions; however, caution should be applied when extrapolating findings from experimental models to meet clinical requirements.

Keywords: anti-sperm antibody, enzyme-linked immunosorbent assay, infertility, testicular sperm aspiration, testosterone.

Association of multilocus sequencing types and antimicrobial resistance profiles of methicillin-resistant Mammaliicoccus sciuri in animals in Southern Thailand

Research (Published online: 15-02-2023)

7. Association of multilocus sequencing types and antimicrobial resistance profiles of methicillin-resistant Mammaliicoccus sciuri in animals in Southern Thailand

Kanpapat Boonchuay, Narin Sontigun, Tuempong Wongtawan, and Punpichaya Fungwithaya

Veterinary World, 16(2): 291-295

ABSTRACT

Background and Aim: Mammaliicoccus sciuri, formerly known as Staphylococcus sciuri, is an opportunistic pathogen in the environment, human and animal mucosa, and skin. Although this pathogen is becoming more resistant to drugs and harmful to animals and humans, basic knowledge of this pathogen remains limited. This study aimed to investigate a new multilocus sequencing type (MLST) related to the antibiotic resistance pattern of M. sciuri from animals in southern Thailand.

Materials and Methods: We used 11 methicillin-resistant M. sciuri (MRMS) isolates in this study which were obtained from six horses, four cows, and one chicken of the previous study. Antimicrobial resistance (AMR) was re-evaluated based on the minimum inhibitory concentration using the VITEK® 2 automated system. Three AMR genes were examined, namely mecA, mecC, and blaZ. Staphylococcal chromosomal cassette mec (SCCmec) gene detection was performed through the multiplex polymerase chain reaction (PCR). Internal segments of the seven housekeeping genes, ack, aroE, ftsZ, glpK, gmk, pta1, and tpiA, were used for multilocus sequence typing. The population of resistant bacteria and the types of multidrug-resistant, extensively drug-resistant, and pandemic drug-resistant bacteria were classified through descriptive analysis.

Results: mecA and blaZ genes were detected in all isolates; however, the mecC gene was not observed in any isolate based on the PCR results. All MRMS isolates revealed a non-typable SCCmec. Seven MLSTs (71, 81, 120, 121, 122, 199, and 200) were identified in this study.

Conclusion: The characteristics of MRMS in Southern Thailand were variable, particularly in cattle and horses. The antibiogram and SCCmec types of this pathogen remain concerns with regard to antibiotic-resistant gene transmission among Staphylococcus and Mammaliicoccus species. All MLSTs in Thailand revealed the distribution among clones in Asia, including the virulence of a zoonotic clone in Southern Thailand.

Keywords: antibiogram, Mammaliicoccus sciuri, methicillin-resistant bacteria, multilocus sequencing types, resistant gene.

First study on capsular serotypes and virulence factors of Pasteurella multocida isolates from Phan Rang sheep in Vietnam

Research (Published online: 14-02-2023)

6. First study on capsular serotypes and virulence factors of Pasteurella multocida isolates from Phan Rang sheep in Vietnam

Phu Van Nguyen, Cong Tuan Le, Xuan Huy Nguyen, Tuan Manh Nguyen, and Kim Cuc Thi Nguyen

Veterinary World, 16(2): 281-290

ABSTRACT

Background and Aim: Pasteurella multocida is considered as a main factor mediating pneumonic pasteurellosis in ruminants, including sheep. It is also a current threat to Phan Rang sheep in Vietnam. This study aimed to characterize P. multocida isolated from Phan Rang sheep, their antibiotic resistance profile, and the prevalence of some virulence-associated genes of these strains.

Materials and Methods: Bacteria were isolated on brain heart infusion, 10% sheep blood agar plates, and screened by biochemical tests. The polymerase chain reaction technique was used with specific primers to identify P. multocida, the presence of virulence-associated genes, and serotypes of isolates. Antimicrobial susceptibility and biofilm formation of isolates were examined using the disk diffusion method and crystal violet-based method, respectively.

Results: A total of 41 P. multocida strains were isolated from 485 samples from clinically sick and healthy sheep. Of the isolates, 58.53% were serotype A, 9.75% were serotype B, and 31.71% were serotype D. Healthy animals were infected with serotype D only. All 15 virulence genes were identified in all strains isolated from clinically sick sheep, while strains isolated from healthy sheep carried 11/15 virulence genes tested. Among virulence-associated genes exbB, exbD, tonB, ompA, oma87, fimA, hgbA, and nanB were detected in over 90% of isolates, whereas hgbB, nanH, tbpA and pfhA were less frequent. Interestingly, pmHAS and tadD were highly prevalent in capsular type A strains, whereas the toxA gene was detected in capsular type D strains only. All of the isolated strains were fully susceptible to enrofloxacin, ciprofloxacin, neomycin, and ofloxacin. About 92.68% were susceptible to chloramphenicol and 90.24% to amikacin, but there was high resistance to erythromycin, tetracycline, and amoxicillin. Our results reveal that 53.65% of 41 isolates could produce biofilm, whereas 46.34% could not.

Conclusion: Pasteurella multocida from Phan Rang sheep possess many virulence genes and resistance to several common antibiotics such as erythromycin, tetracycline, and amoxicillin. The results are an important warning regarding antibiotic resistance of P. multocida.

Keywords: antimicrobial resistance, biofilm formation, capsular type, Pasteurella multocida, Phan Rang sheep.

Whole-genome sequencing of Histophilus somni strains isolated in Russia

Research (Published online: 14-02-2023)

5. Whole-genome sequencing of Histophilus somni strains isolated in Russia

Svetlana Yatsentyuk, Julia Pobolelova, Veronika Gordeeva, and Irina Timofeeva

Veterinary World, 16(2): 272-280

ABSTRACT

Background and Aim: Histophilus somni is a Gram-negative bacterium belonging to the Pasteurellaceae family that can cause bovine histophilosis. Histophilus may act as a commensal or opportunistic bacterial cattle pathogen. Comparing genomes of the pathogenic strain 2336 with the non-pathogenic preputial 129Pt isolate revealed some putative virulence factors. The study of the complete genomes of H. somni strains circulating in Russia has never been conducted before. This study aimed to identify genetic features of the H. somni strains isolated in Russia and evaluate the possibility of using strains for vaccine development.

Materials and Methods: Three strains of H. somni were isolated from different sources. Strain 188-VIEV was isolated from a vaginal swab sample of cattle with endometritis. 532-VIEV and 551-VIEV were cultured from the cryopreserved bull semen samples imported from Canada. Histophilus somni strain ATCC 700025 provided by ATCC (American Type Culture Collection) was also used in the study. DNA extraction was performed using QIAamp DNA Mini Kit (QIAGEN, USA). The whole-genome sequencing of the four strains was performed using Illumina Miseq. The comparison of the resulting sequences with the complete genomes of H. somni 2336 and 129Pt, and detection of the resistance genes and virulence factors, was performed using the ResFinder and Virulence Factor Database web services.

Results: The genome size of the samples varied from 1.9 to 2.3 Mb. The number of coding sequences varied from 1795 to 2256. The average sequence density was 90%. The total guanine-cytosine (GC) content was 36.8%–37.2%, which coincided with data previously obtained for H. somni. Three out of four studied strains encoded putative virulence factors such as filamentous hemagglutinin homologs, lipooligosaccharide biosynthesis proteins, and proteins involved in iron transport and utilization. The Ser83Ile substitution was identified in the DNA topoisomerase II (gyrA) in H. somni strains 532-VIEV and 551-VIEV cultured from bull semen which led to resistance to fluoroquinolones. The gene (AAC-6-Ia + APH-2”) encoding a bifunctional aminoglycoside modification enzyme was detected in strain 551-VIEV.

Conclusion: Strains with virulence genes identified could be candidates for designing vaccines and potentially represent antigen sources. The results show that antibiotic-resistant H. somni can be spread with semen used for artificial insemination.

Keywords: antibiotic resistance genes, Histophilus somni, next-generation sequencing, virulence factors.

Antimicrobial susceptibility test and antimicrobial resistance gene detection of extracellular enzyme bacteria isolated from tilapia (Oreochromis niloticus) for probiotic candidates

Research (Published online: 11-02-2023)

4. Antimicrobial susceptibility test and antimicrobial resistance gene detection of extracellular enzyme bacteria isolated from tilapia (Oreochromis niloticus) for probiotic candidates

Mira Mawardi, Agustin Indrawati, I. Wayan Teguh Wibawan, and Angela Mariana Lusiastuti

Veterinary World, 16(2): 264-271

ABSTRACT

Background and Aim: Antimicrobial resistance (AMR) is a global problem that can increase mortality and morbidity rates and adversely affect health. Therefore, AMR control must be carried out in various sectors, including the fisheries sector, using probiotics. Bacteria can become resistant to antibiotics, including bacteria used for probiotics. This study aimed to isolate bacteria as potential producers of extracellular enzymes, phenotypic characterization, and antibiotic-resistant gene patterns.

Materials and Methods: In this study, 459 bacterial isolates were isolated from the stomach of tilapia in Indonesia. Tilapia was obtained from Sukabumi, Ciamis, Serang, Banjarnegara, Jayapura, Sorong, Manokwari Selatan, Takalar, Lampung, Batam, and Mandiangin. Enzymatic bacteria were identified. An antimicrobial susceptibility test was conducted by agar disk diffusion, and genotypic detection of encoding genes was performed using a molecular method.

Results: This study obtained 137 isolates (29.84%) that can produce extracellular enzymes. The highest number of E-sensitive isolates was found, including 130 isolates (94.89%). Six isolates (6/137) can produce four enzymes (amylase, protease, cellulose, and lipase), and they were sensitive to antibiotics. A total of 99 isolates can produce extracellular enzymes, and they were sensitive to antibiotics. Such isolates serve as a consortium of probiotic candidates. The isolates that are resistant to oxytetracycline (OT), erythromycin (E), tetracycline (TE), and enrofloxacin (ENR) included 15 isolates (10.95%), seven isolates (5.11%), three isolates (2.19%), and one isolate (0.73%), respectively. In addition, four isolates (2.92%) were detected as multidrug-resistant. The tet(A) gene obtained the highest result of detection of resistance genes in isolates that were intermediate and resistant to TE and OT. Isolates that serve as ENR intermediates have a high qnr(S) resistance gene.

Conclusion: The data in this study provide the latest update that bacteria can serve as a consortium of potential probiotics with antibiotic-resistant genes for the treatment of fish. Bacteria that are intermediate to antibiotics may contain resistance genes. The results of this study will improve the policy of probiotic standards in Indonesia.

Keywords: antibiotic, antimicrobial resistance, enzymatic bacteria, Oreochromis niloticus, probiotic.