Hoplobatrachus Rugulosus Research Articles

<span>Some data about the food composition of amphibians in Cao Lanh city, Dong Thap province </span>

On the rice field ecosystem in Cao Lanh district, Dong Thap province, there are 6 species of amphibians distributed: Duttaphrynus melanostictus, Kaloula pulchra and Hylarana guentheri have the most diversity of feed ingredients; Fejervarya limnocharis, Hoplobatrachus rugulosus and Ichthyophis bannanicus have less food diversity; Scolopendomorpha and Polydesmoidea are the least, accounting for 1,2%. Amphibians have the most food frequency in Coleoptera, followed by Hymenoptera and Orthoptera. Amphibians are usually terrestrial, eating mainly insects. The frequency of pests was lowest in the period from sowing seeds until the rice had 3 leaves germinated, the lowest in the period of yellow rice until after harvest, the highest at the stage of rice with milk in the grain to the stage of green - ripened rice. The frequency of encounters of Hoplobatrachus rugulosus and Fejervarya limnocharis is lowest at the stage of seeding until rice germination, decrease gradually in the period of ripe rice until after harvest, stage 4 (Rice with milk in the grain until green ripening) has the highest frequency. In stage 1, there is the lowest frequency of encountering amphibians and pests, frequency increases gradually to stage 2 and peaks at stage 4, decreasing at stage 5.

Intraocular Pressure in Chinese Edible Frogs (Hoplobatrachus ruguosus) by Rebound Tonometry: Comparing Sex and Use of Anesthesia

Abstract The objective of this study was to describe the intraocular pressure (IOP) findings in a population of Chinese edible frogs (Hoplobatrachus rugulosus) sold in wet markets across Hong Kong and to characterize the influence of sex and anesthesia. One hundred animals were evaluated and ninety-six were included in this study. All frogs included appeared ophthalmologically normal upon gross examination. Intraocular pressure was evaluated with rebound tonometry (TonoVet Plus® setting “d”) on c conscious frogs and under anesthesia. General anesthesia was induced with MS-222 (tricaine methanesulfonate) 2 g/L by immersion. After IOP measurements, the frogs were euthanized with intravenous pentobarbital (200 mg/ml) and sex was confirmed on postmortem examination. Mean (+/− SD) IOP values were significantly decreased under MS-222 anesthesia in males (14.7 +/− 2.8 mmHg; P < 0.0001) and females (14.7 +/− 2.4 mmHg; P < 0.0001) compared to conscious readings (16.3 +/− 2.1 mmHg and 16.3 +/− 2.1 mmHg, respectively). There was no significant difference in IOP found between males and females before or after anesthesia. There was no significant difference between the right and left eye in males and females before or after anesthesia. The results of this study establish a normal range of IOP for this species and are comparable to previous published reports of IOP in anurans.

Effects of Mixed Mineral Supplementation in On-Farm Feeds on the Metamorphosis Development and Growth Performances of Frog (Hoplobatrachus rugulosus) Tadpoles

Effects of Mixed Mineral Supplementation in On-Farm Feeds on the Metamorphosis Development and Growth Performances of Frog (Hoplobatrachus rugulosus) Tadpoles

Parasitic helminths of alien invasive anurans in Butuan City, Northeastern Mindanao, Philippines.

This study aimed to identify the helminth parasites of invasive anuran species in selected barangays in Butuan City, Philippines. In urbanized areas, invasive species dominate anuran diversity, and one of the primary threats they pose to native wildlife is the transmission of diseases and parasites. Out of the 91 collected individuals of invasive anuran species, Rhinella marina was the most abundant (88 %), followed by Hoplobatrachus rugulosus (12 %) and Kaloula pulchra (3 %). The study identified five species of parasites, with Spirometra sp. being the most prevalent (17.58 %), followed by Echinostoma sp. (16.5 %), Rhabdias bufonis (14.3 %), Cosmocerca sp. (6.6 %), and Strongyloides stercoralis (3.30 %), respectively. Spirometra sp. also had the highest intensity (7.67), followed by Cosmocerca sp. (5), Strongyloides stercoralis (3.33), Rhabdias bufonis (3.30), and Echinostoma sp. (2.73). This parasitological survey revealed that H. rugulosus had the highest prevalence and infection of parasites, and residential areas had the highest parasite prevalence among the habitat types. Adult hosts were found to harbor a higher prevalence and intensity, and male hosts had a higher prevalence. The results highlight the high risk of parasite transmission from anurans to other animals and emphasize the need for the community to control the population of invasive anuran species for the safety of native anurans and to prevent zoonotic transmission to other animals and humans.

Peptide TK-HR from the Skin of Chinese Folk Medicine Frog Hoplobatrachus Rugulosus Accelerates Wound Healing via the Activation of the Neurokinin-1 Receptor.

Wound healing is a complex process and remains a considerable challenge in clinical trials due to the lack of ideal therapeutic drugs. Here, a new peptide TK-HR identified from the skin of the frog Hoplobatrachus rugulosus was tested for its ability to heal cutaneous wounds in mice. Topical application of TK-HR at doses of 50-200 μg/mL significantly accelerated wound closure without causing any adverse effects in the animals. In vitro and in vivo investigations proved the regulatory role of the peptide on neutrophils, macrophages, keratinocytes, and vein endothelial cells involved in the inflammatory, proliferative, and remodeling phases of wound healing. Notably, TK-HR activated the MAPK and TGF-β-Smad signaling pathways by acting on NK1R in RAW264.7 cells and mice. The current work has identified that TK-HR is a potent wound healing regulator that can be applied for the treatment of wounds, including diabetic foot ulcers and infected wounds, in the future.

Steroidogenic potential of the gonad during sex differentiation in the rice field frog Hoplobatrachus rugulosus (Anura: Dicroglossidae).

Prior studies demonstrated that gonadal differentiation in the rice field frog, Hoplobatrachus rugulosus, was of an undifferentiated type since all individuals had ovaries at complete metamorphosis. However, the steroidogenic potential of the gonad is still unknown. In this study, H. rugulosus were obtained by stimulating fertilization in the laboratory under natural light and temperature conditions. The gonads were collected and their steroidogenic potential was evaluated by determining the expression level of messenger RNA (mRNA) encoding for cytochrome P450 17-hydroxylase/C17-20 lyase (CYP17) and cytochrome P450 aromatase (CYP19) using quantitative real-time RT-PCR and the localization of CYP17 mRNA in tissues by in situ hybridization. The CYP17 mRNA levels in males at 4-11 weeks postmetamorphosis were higher than in female and intersex gonads. This corresponded to their localization in the gonadal tissues, where CYP17 signals were specifically detected in the Leydig cells of the testis at 5-16 weeks postmetamorphosis but was undetectable in all ovary samples. The CYP19 mRNA levels in females at 4-11 weeks postmetamorphosis was higher than in male and intersex gonads, which corresponded with gonadal development, indicating the potential steroidogenic function of the ovary. Based on the present results, the role of CYP17 and CYP19 mRNA in sex differentiation in H. rugulosus may occur after gonadal sex differentiation and the steroidogenic potential of the gonads exhibited a sexual dimorphic pattern. These results provide a crucial basis for further research on the developmental biology in anuran species.

Serotypes, Antimicrobial Resistance Profiles, and Virulence Factors of Salmonella Isolates in Chinese Edible Frogs (Hoplobatrachus rugulosus) Collected from Wet Markets in Hong Kong.

Salmonella is an important agent of gastrointestinal disease in humans. While livestock, such as cattle, poultry, and pigs, are well-recognised animal reservoirs of Salmonella, there is a lack of data on Salmonella in edible frogs, even though frog meat is a popular food worldwide. In this study, 103 live edible Chinese frogs (Hoplobatrachus rugulosus) were collected from wet markets throughout Hong Kong. After euthanasia, faeces or cloacal swabs were examined for Salmonella. Overall, Salmonella spp. were isolated from 67 (65%, CI: 0.554-0.736) of the samples. The serotypes included S. Saintpaul (33%), S. Newport (24%), S. Bareilly (7%), S. Braenderup (4%), S. Hvittingfoss (4%), S. Stanley (10%), and S. Wandsworth (16%). Many isolates were phylogenetically related. A high number of genes encoding for resistance to clinically relevant antimicrobials, and a high number of virulence determinants, were identified. Antimicrobial susceptibility testing (AST) identified multidrug resistance (MDR) in 21% of the isolates. Resistance to ampicillin, ciprofloxacin, nalidixic acid, and tetracycline was common. These results demonstrate that a high percentage of live frogs sold for human consumption in wet markets are carriers of multidrug-resistant Salmonella. Public health recommendations for handling edible frogs should be considered, to mitigate the risk of Salmonella transmission to humans.

The first Brevinin-1 antimicrobial peptide with LPS-neutralizing and anti-inflammatory activities in vitro and in vivo.

Antimicrobial peptide is one important component of the first protective barrier of organisms. They not only have potent antimicrobial activity which can protect the body from the invading pathogens, but also participate in the immune regulation of the body. In this study, a Brevinin-1 peptide named by Brevinin-1GHd was identified from Hoplobatrachus rugulosus, and the similarity of mature peptide sequence among Brevinin-1GHd, Brevinin-1HL and Brevinin-1GHa supported the close species relationship between H. rugulosus, Hylarana latouchii and Hylarana guertheri. Moreover, the secondary structure of Brevinin-1GHd was found to possess α-helical characteristics and high thermal stability. In addition, Brevinin-1GHd could bind to LPS with a Kd value of 6.49 ± 5.40 mM and suppress the release of TNF-α, NO, IL-6 and IL-1β by inactivation of MAPK signaling pathway in RAW 264.7 cells induced by LPS. Furtherly, Brevinin-1GHd had a significant inhibitory effect on acute edema development in the right paw of mice injected by carrageenan. Thus, the significant LPS-neutralizing and anti-inflammatory activities of Brevinin-1GHd were demonstrated in this study, which made it become the first Brevinin-1 family peptide with anti-inflammatory activity reported so far, and the biological activity of Brevinin-1GHd made it promising to be a novel therapeutic drug for infectious inflammation.

Dietary preference of Alien Invasive Anurans (Rhinella marina, Hoplobatrachus rugulosus, and Kaloula pulchra) in Butuan City, Northeastern Mindanao, Philippines

Invasive anuran species (IAS) threaten the native anuran species as they compete for food sources. The diet of the three IAS, Rhinella marina, Kaloula pulchra, and Hoplobatrachus rugulosus, were collected from different localities in Butuan City. The gut collected from 91 individuals of IAS, i.e., R. marina (n=80), H. rugulosus (n=10), and K. pulchra (n=1) were examined for food items using a dissecting microscope. The R. marina had the most recorded prey items, consisting of plant materials, pebbles, insects, and reptile-like species, while K. pulchra prefers small and slow-moving insects. Hoplobatrachus rugulosus, which also preferred insects on their diet, exhibited frog predation. Among the food categories, plant materials (56%) had the highest percentage, followed by invertebrates (47%), other organic and inorganic materials (12%), and vertebrates (1%). Formicidae, Coleoptera, and Odonata have the highest percentage among the invertebrate prey types. The assessed amphibians feed on locally available prey, generally insects, and suggest potential competition for food resources with native anuran populations.

First Report on the Presence of Toxic Metals and Metalloids in East Asian Bullfrog (Hoplobatrachus rugulosus) Legs.

We examined the presence of As, Cr, Cd, and Pb in 42 samples of farmed East Asian bullfrog (Hoplobatrachus rugulosus) from Vietnam and Thailand by inductively coupled plasma-mass spectrometry (ICP-MS). An estimation of the dietary intake and exposure to the toxic elements analysed was also carried out. The results showed very high As levels, with mean values of 0.094 ± 0.085 mg/Kg w.w. and a maximum of 0.22 mg/Kg. No significant differences were found for As contents between areas of production (p > 0.05). No detectable Cd contents were found in all the samples examined. The Pb concentrations of the East Asian bullfrog legs samples were below the European Commission’s permitted levels. The Cr and Pb contents of the East Asian bullfrog produced in Vietnam were significantly higher than that produced in Thailand (p < 0.05). The target hazard quotient (THQ) ratio for Cr was not exceeded for all the samples analysed. In contrast, the benchmark dose lower confidence limit (BMDL) and THQ ratios for As were exceeded, indicating carcinogenic and non-carcinogenic risks for those who consume this type of food. The results of this work confirm the role of As-contaminated water absorption as an important source of arsenic for these adult organisms.

Anurans of select green spaces of Davao City, Mindanao Island, Philippines

Abstract. Delima-Baron EM, Ruales CAS, Tripole C. Tagoon MDT, Susulan TB. 2022. Anurans of select green spaces of Davao City, Mindanao Island, Philippines. Biodiversitas 23: 4810-4815. Data on anurans in green spaces, especially in highly urbanized cities, including Davao, is still very limited. We present for the first time a checklist of anuran species from six green spaces of Davao City previously not surveyed for anuran data. Anurans were documented from January to April 2019 using visual encounter techniques coupled with microhabitat searches on several nocturnal surveys. Eighteen species representing seven families were recorded. Three endemic species: Pelophryne brevipes, Fejervarya moodiei, and F. vittigera, were added to the current list of anurans known from green spaces of Davao City, bringing the total known species for Davao City to 23. Thirteen of the species recorded are endemic. Two species under the Near Threatened category of IUCN, Limnonectes magnus and Sanguirana everetti, were also documented. The study also accounted for four invasive species: Rhinella marina, Eleutherodactylus planirostris, Hoplobatrachus rugulosus, and Kaloula pulchra. However, these species were encountered only in sites with artificial structures nearby, prominent noise, and has less heterogeneous vegetation. Data adds to the limited account of anurans in Davao City’s green spaces. Adding more species also implies that the list of anurans from Davao City’s green spaces is far from complete. Exploration of other green spaces available in the city is necessary to fully understand anuran species composition and the value of green spaces to the persistence of this taxon even in highly urbanized areas.

Cathelicidin-HR from Hoplobatrachus rugulosus: an antioxidant peptide that performs a protective effect against UV/H2O2 -induced DNA damage

Cathelicidin-HR from Hoplobatrachus rugulosus: an antioxidant peptide that performs a protective effect against UV/H2O2 -induced DNA damage

Mitochondrial gene expression in different organs of Hoplobatrachus rugulosus from China and Thailand under low-temperature stress

BackgroundHoplobatrachus rugulosus (Anura: Dicroglossidae) is distributed in China and Thailand and the former can survive substantially lower temperatures than the latter. The mitochondrial genomes of the two subspecies also differ: Chinese tiger frogs (CT frogs) display two identical ND5 genes whereas Thai tiger frogs (TT frogs) have two different ND5 genes. Metabolism of ectotherms is very sensitive to temperature change and different organs have different demands on energy metabolism at low temperatures. Therefore, we conducted studies to understand: (1) the differences in mitochondrial gene expression of tiger frogs from China (CT frogs) versus Thailand (TT frogs); (2) the differences in mitochondrial gene expression of tiger frogs (CT and TT frogs) under short term 24 h hypothermia exposure at 25 °C and 8 °C; (3) the differences in mitochondrial gene expression in three organs (brain, liver and kidney) of CT and TT frogs.ResultsUtilizing RT-qPCR and comparing control groups at 25 °C with low temperature groups at 8 °C, we came to the following results. (1) At the same temperature, mitochondrial gene expression was significantly different in two subspecies. The transcript levels of two identical ND5 of CT frogs were observed to decrease significantly at low temperatures (P < 0.05) whereas the two different copies of ND5 in TT frogs were not. (2) Under low temperature stress, most of the genes in the brain, liver and kidney were down-regulated (except for COI and ATP6 measured in brain and COI measured in liver of CT frogs). (3) For both CT and TT frogs, the changes in overall pattern of mitochondrial gene expression in different organs under low temperature and normal temperature was brain > liver > kidney.ConclusionsWe mainly drew the following conclusions: (1) The differences in the structure and expression of the ND5 gene between CT and TT frogs could result in the different tolerances to low temperature stress. (2) At low temperatures, the transcript levels of most of mitochondrial protein-encoding genes were down-regulated, which could have a significant effect in reducing metabolic rate and supporting long term survival at low temperatures. (3) The expression pattern of mitochondrial genes in different organs was related to mitochondrial activity and mtDNA replication in different organs.

Microbial Diversity of the Chinese Tiger Frog (Hoplobatrachus rugulosus) on Healthy versus Ulcerated Skin.

Simple SummaryAs amphibians’ skin is highly sensitive to the environment, skin defects such as ulceration may pose a particular threat to them. Our study has found a stark difference in the microbial communities between healthy and ulcerated Hoplobatrachus rugulosus skin. The proportion and type of bacteria differed between the two groups, and we suggest that ulceration on the skin may lead to changes in skin microbial communities. The functional pathways of skin microbes may be influenced by ulceration on the skin surface of H. rugulosus. We also found that Vogesella is more abundant in healthy H. rugulosus, which may be a potential probiotic candidate for the reduction or removal of pathogens.The Chinese tiger frog (Hoplobatrachus rugulosus) is extensively farmed in southern China. Due to cramped living conditions, skin diseases are prevalent among unhealthy tiger frogs which thereby affects their welfare. In this study, the differences in microbiota present on healthy versus ulcerated H. rugulosus skin were examined using 16S rRNA sequences. Proteobacteria were the dominant phylum on H. rugulosus skin, but their abundance was greater on the healthy skin than on the ulcerated skin. Rhodocyclaceae and Comamonadaceae were the most dominant families on the healthy skin, whereas Moraxellaceae was the most dominant family on the ulcerated skin. The abundance of these three families was different between the groups. Acidovorax was the most dominant genus on the healthy skin, whereas Acinetobacter was the most dominant genus on the ulcerated skin, and its abundance was greater on the ulcerated skin than on the healthy skin. Moreover, the genes related to the Kyoto Encyclopedia of Genes and Genomes pathways of levels 2–3, especially those genes that are involved in cell motility, flagellar assembly, and bacterial chemotaxis in the skin microbiota, were found to be greater on the healthy skin than on the ulcerated skin, indicating that the function of skin microbiota was affected by ulceration. Overall, the composition, abundance, and function of skin microbial communities differed between the healthy and ulcerated H. rugulosus skin. Our results may assist in developing measures to combat diseases in H. rugulosus.

Mercy or messy: distribution and differentiation of native and released Chinese bullfrogs (Hoplobatrachus rugulosus) in Hong Kong using genetic and morphological analyses

Mercy or messy: distribution and differentiation of native and released Chinese bullfrogs (Hoplobatrachus rugulosus) in Hong Kong using genetic and morphological analyses

Ecotoxicity assessment of triphenyl phosphate (TPhP) exposure in Hoplobatrachus rugulosus tadpoles

Triphenyl phosphate (TPhP), a widely used aromatic organophosphate flame retardant, is known to accumulate in organisms through water, air, and soil, consequently, causing toxicity. This study is the first to evaluate the acute and sub-chronic toxicities of TPhP to amphibians. In the acute toxicity analysis, the 96-h median lethal concentration (LC50) for GS35 Hoplobatrachus rugulosus tadpoles was 2.893 mg/L, and the 10% effect concentration (EC10) was 289 μg/L. After two weeks of exposure to low TPhP concentrations, the survival and metamorphosis rates of H. rugulosus tadpoles decreased, and the metamorphosis time was prolonged as the TPhP concentration increased. The threshold concentration that affected tadpole survival and metamorphosis time was 50 μg/L and 100 μg/L, respectively. No significant differences were observed in the condition factor and hepatic somatic index of the tadpole after metamorphosis; however, tadpole body mass and TPhP concentration were negatively correlated. Further, TPhP inhibited the expressions of Cu–Zn sod and cat, thereby reducing the activities of superoxide dismutase and catalase in the tadpole liver. The threshold for affecting gene expression and enzymatic activity was 100 μg/L. These findings provide significant insights on the stress ecology of aquatic organisms.

Genome-Wide SNP Analysis of Male and Female Rice Field Frogs, Hoplobatrachus rugulosus, Supports a Non-Genetic Sex Determination System

Sex determination systems (SDSs) in anurans are diverse and have undergone independent evolutionary transitions among species. The mode of sexual reproduction of the rice field frog (Hoplobatrachus rugulosus)—an economically viable, edible amphibian species—is not well known. Previous studies have proposed that threshold temperature conditions may determine sex in these frogs. To elucidate the SDS in H. rugulosus, we karyotyped 10 male and 12 female frogs, and performed fluorescence in situ hybridization combined with sequencing analyses using DArTseq™. Our results revealed a highly conserved karyotype with no sex chromosome heteromorphism, and the sequencing analyses did not identify any consistent sex-linked loci, supporting the hypothesis of temperature-dependent sex determination. The results of this study, and others, on SDSs in the rice field frog and related species also provide support for the theory that heteromorphic sex chromosomes may lead to an evolutionary trap that prevents variable SDSs. These findings add important information to the body of knowledge on H. rugulosus and are likely to have a significant impact on the productivity and economic success of rice field frog farming.

Detection of Gnathostoma spinigerum Advanced 3rd-Stage Larvae in the Chinese Edible Frog, Hoplobatrachus rugulosus, from Local Markets in Phnom Penh, Cambodia.

The Chinese edible frogs, Hoplobatrachus rugulosus (n=20), and the striped snakehead fish, Channa striata (n=34), were purchased from local markets in 3 administrative regions of Cambodia (Phnom Penh, Pursat, and Takeo Provinces) from May 2017 to April 2019, and their infection status with Gnathostoma sp. larvae was investigated. The frogs and fish were transported to the laboratory with ice and examined using the artificial digestion method. Advanced 3rd-stage larvae (AdL3) of Gnathostoma spinigerum, 24 in total number (1–6 larvae/frog), were detected from 6 (60.0%) out of 10 frogs purchased from Phnom Penh. No gnathostome larvae were detected in 10 frogs purchased from Takeo Province and 34 snakeheads from Phnom Penh, Pursat, and Takeo Provinces. AdL3 isolated from the frogs were 2.55–3.90 mm long and 0.31–0.36 mm wide. They had a characteristic head bulb (0.081×0.191 mm in average size) with 4 rows of hooklets, a muscular long esophagus (0.950–1.230 mm long), and 2 pairs of cervical sacs (0.530–0.890 mm long). The average number of hooklets in the 1st, 2nd, 3rd, and 4th rows was 41, 45, 48, and 51, respectively. These features were consistent with G. spinigerum AdL3. By the present study, it has been first confirmed that the Chinese edible frog, H. rugulosus, from Phnom Penh serves as a second intermediate host for G. spinigerum, although their intensity of infection was not so high compared to other previously reported localities.

Analyzing the gonadal transcriptome of the frog Hoplobatrachus rugulosus to identify genes involved in sex development

BackgroundThe tiger frog (Hoplobatrachus rugulosus) is listed as a national Class II protected species in China. In the context of global warming, the sex ratio of amphibians will be affected, and the development of the population will be limited. Therefore, considering the potential for a decrease in the number of amphibians, studying sex evolution and molecular regulation of gonadal development in H. rugulosus, phenomenon that are currently unclear, is of great significance.ResultsHere, H. rugulosus was used to explore the mechanisms regulating gonadal development in amphibians. Illumina HiSeq 3000 was used to sequence the gonadal transcriptome of male and female H. rugulosus at two growth stages to identify genes related to gonadal development and analyze expression differences in the gonads. This analysis indicated that cyp17α, hsd3β, hsd11β1, cyp19α, and hsd17β12 perform vital functions in sex development in amphibians. Specifically, the expression of cyp3α, cyp17α, hsd3β, hsd11β1, sox2, sox9, sox30, soat, cyp19α, hsd17β12, and hspα1s was correlated with gonadal development and differentiation in H. rugulosus, as determined using the quantitative reverse transcriptase-polymerase chain reaction.ConclusionSignificant differences were found in the gonadal gene expression levels in H. rugulosus of both sexes, and we identified a steroid hormone synthesis pathway in this species and analyzed related gene expression, but the changes during sex differentiation were still unclear. To our knowledge, this report presents the first analysis of the H. rugulosus gonadal transcriptome and lays the foundation for future research.

Dietary Spirulina (Arthrospira platensis) Supplementation on Growth Performance, Haematology, Immune Response and Disease Resistance of Rugose Frog (Hoplobatrachus rugulosus)

The present study aimed to investigate the effects of dietary Spirulina (Arthrospira platensis) supplementation on the growth, survival, haematology, immune parameters and disease resistance of Rugose frog (Hoplobatrachus rugulosus) against Aeromonas hydrophila infection. Frogs were fed a formulated diet containing Spirulina as 0% (control), 1.5%, 3.0%, 4.5%, and 6.0% for a period of eight weeks. The results indicated that growth parameters; final body weight, weight gain, average daily gain, specific growth rate, feed conversion ratio, and survival rate of frog fed with A. platensis at the level of 4.5% showed best values but not significantly different (P&gt;0.05). However, haematological values; leukocytes, erythrocytes, haemoglobin, and haematocrit levels were highest and significantly different (P&lt;0.05) when compared to the control group. Disease resistance was tested by challenging with A. hydrophila after eight weeks of feeding. The results showed that frog fed with 4.5% A. platensis showed significantly (P&lt;0.05) higher survival rate and highest resistance to A. hydrophila in comparison with the other groups. Leukocyte, erythrocyte, complement C3, and immunoglobulin G levels of frogs fed with 4.5% A. platensis were found to be significantly (P &lt; 0.05) differed compared with control. In conclusion, diet supplemented with Spirulina at the level of 4.5% could improve growth performance, haematological and immune parameters, and A. hydrophila resistance in Rugose frogs.