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Fulltext Prevalence, antibiotic susceptibility, and presence of drug resistance genes in Aeromonas spp. isolated from freshwater fish in Kelantan and Terengganu states, Malaysia

Fauzi NNFNM, Hamdan RH, Mohamed M, Ismail A, Mat Zin AA, Mohamad NFA

Vet World, 2021 Aug;14(8):2064-2072.
PMID: 34566322 DOI: 10.14202/vetworld.2021.2064-2072

Background and Aim: The emergence of antibiotic-resistant bacterial pathogens has been increasingly reported, which has resulted in a decreasing ability to treat bacterial infections. Therefore, this study investigated the presence of Aeromonas spp., including its antibiotic resistance in various fish samples, Oreochromis spp., Clarias gariepinus, and Pangasius hypophthalmus, obtained from Kelantan and Terengganu, Malaysia.
Materials and Methods: In this study, 221 fish samples, of which 108 (Oreochromis spp., n=38; C. gariepinus, n=35; and P. hypophthalmus, n=35) were from Kelantan and 113 (Oreochromis spp., n=38; C. gariepinus, n=35; and P. hypophthalmus, n=40) were from Terengganu, were caught using cast nets. Then, samples from their kidneys were cultured on a Rimler Shott agar to isolate Aeromonas spp. Polymerase chain reaction (PCR) was used to confirm this isolation using specific gene primers for species identification. Subsequently, the isolates were tested for their sensitivity to 14 antibiotics using the Kirby-Bauer method, after which the PCR was conducted again to detect resistance genes: sul1, strA-strB, aadA, bla TEM, bla SHV, tetA-tetE, and tetM.
Results: From the results, 61 isolates were identified as being from the genus Aeromonas using PCR, of which 28 were Aeromonas jandaei, 19 were Aeromonas veronii, seven were Aeromonas hydrophila, and seven were Aeromonas sobria. Moreover, 8, 12, and 8 of A. jandaei; 4, 3, and 12 of A. veronii; 6, 0, and 1 of A. hydrophila; and 3, 3, and 1 of A. sobria were obtained from Oreochromis spp., C. gariepinus, and P. hypophthalmus, respectively. In addition, the isolates showed the highest level of resistance to ampicillin (100%), followed by streptomycin (59.0%), each kanamycin and nalidixic acid (41.0%), neomycin (36.1%), tetracycline (19.7%), sulfamethoxazole (14.8%), and oxytetracycline (13.1%). Resistance to gentamicin and ciprofloxacin both had the same percentage (9.8%), whereas isolates showed the lowest resistance to norfloxacin (8.2%) and doxycycline (1.6%). Notably, all Aeromonas isolates were susceptible to chloramphenicol and nitrofurantoin. Results also revealed that the multiple antibiotic resistances index of the isolates ranged from 0.07 to 0.64, suggesting that the farmed fish in these areas were introduced to the logged antibiotics indiscriminately and constantly during their cultivation stages. Results also revealed that the sul1 gene was detected in 19.7% of the Aeromonas isolates, whereas the tetracycline resistance genes, tetA and tetE, were detected in 27.9% and 4.9% of the isolates, respectively. However, β-lactam resistance genes, bla TEM and bla SHV, were found in 44.3% and 13.1% of Aeromonas isolates, respectively, whereas strA-strB and aadA genes were found in 3.3% and 13.1% of the isolates, respectively.
Conclusion: This study, therefore, calls for continuous surveillance of antibiotic-resistant Aeromonas spp. in cultured freshwater fish to aid disease management and better understand their implications to public health.
Detection of pathogenic Vibrio species and antibiogram activity in Asian Seabass (Lates calcarifer) in Tumpat, Kelantan

Auzureen AMZ, Michael MS, Mohamed M, Peng TL, Fauzi F, Mohamad NFA, et al.

Trop Biomed, 2022 Dec 01;39(4):569-574.
PMID: 36602217 DOI: 10.47665/tb.39.4.013

Some of Vibrio species is well known as pathogenic bacteria in aquaculture and the marine industry. Its infection is able to generate a massive outbreak and affect the fish population, especially for net caged fish such as seabass. This study was conducted to investigate the prevalence of Vibrio spp. isolated from seabass (Lates calcarifer) in Sri Tujuh Lagoon, Tumpat, Kelantan. Then, to determine the antibiotic resistance in Vibrio isolates. Polymerase chain reaction (PCR) was used to detect Vibrio species using specific primer VR169 and VR744 with estimation base pair size band, 597 bp and further identified by sequencing. On the other hand, antibiotic susceptibility tests were continued by using 13 types of antibiotics; kanamycin (K30), chloramphenicol (C30), neomycin (N10), ampicillin (AMP10), nitrofurantoin (F300), tetracycline (TE30), streptomycin (S10), norfloxacin (NOR10), ciprofloxacin (CIP5), nalidixic acid (NA30), gentamicin (CN10), doxycycline (DO30) and sulfamethoxazole (SXT100). As a result, 14 Vibrio isolates were identified, including Vibrio fluvialis (n=6), Vibrio parahaemolyticus (n=3), Vibrio harveyi (n=2) and each isolate for Vibrio vulnificus, Vibrio alginolyticus and Vibrio spp. The results showed that all isolates were sensitive to most antibiotics except ampicillin, neomycin and streptomycin. The MAR index value was ranging from 0 to 0.31. This study demonstrates the prevalence of Vibrio spp. in seabass and the report on multidrug resistance strains that could be of concern to the fish farmers. In addition, data from this study can be further used in fish disease management plans.
First report of Anaplasma marginale and Anaplasma ovis in goats in Kelantan, Malaysia

Peng TL, Yahya SNC, Mohamed M, Hamdan RH, Shean CS, Okene IA, et al.

Trop Biomed, 2024 Dec 01;41(4):404-411.
PMID: 39876496 DOI: 10.47665/tb.41.S.004

Anaplasma species are obligate rickettsial intraerythrocytic pathogens that cause an important tick-borne disease of economic importance in livestock production in many countries. Anaplasma species have been detected from farm animals worldwide, there is a paucity of information on Anaplasma infections in goats from Malaysia. Thus, this study aimed to assess the infection rate and identify Anaplasma species and some selected risk factors in goats across selected districts in Kelantan, Malaysia. A total of 411 blood samples were collected from goats and analysed for Anaplasma species targeting the msp4 gene using conventional PCR and sequencing. The infection risk was determined by breed, age, management system and location. Our results indicate an overall infection rate of 30.9% for Anaplasma species detected. Interestingly, sequencing of selected amplicons revealed the presence of Anaplasma ovis and A. marginale. Data analysis revealed a marked statistically significant association between Anaplasma infection and some variables such as location (district), farm management system, breed, and age (P < 0.05). Specifically, goats raised on intensive management had the highest prevalence of 46.25% (37/80) compared to other management types. Also, with regards to district, goats raised in the coastal region had a higher prevalence of 39.23% (71/181) compared to those raised in inland region 24.35% (56/230). Regarding breed, goats that were of the pure breed had a higher prevalence of Anaplasma species infection 38.19% (97/254) compared to crossbreeds with a prevalence of 19.11% (30/157). Lastly, goats 3 years had the least prevalence 18.99% (34/179). To the best of our knowledge, this is the first report of A. marginale and A. ovis in goats from northeastern Peninsular Malaysia. The infected goats were clinically healthy; this revealed the role of goats as a potential reservoir for A. marginale and the presence of A. ovis in goats in Malaysia. Continuous efforts towards tick control must be sustained to ensure high productive yield and reduced disease burden associated with TBPs of goats in the study area.

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