Ascorbic acid influences the development and immunocompetence of larval Heliothis virescens

Ascorbic acid, known to be a free radical scavenger and vital to insect development, is important in larval resistance to baculovirus infection. We sequentially elevated the ascorbic acid content in an ascorbic acid-depleted diet and evaluated the effect on larval Heliothis virescens (Fabricius) (Lepidoptera: Noctuidae) development and immunocompetence. Dietary ascorbic acid levels lower than 0.7 g l⁻¹ slowed the growth rate of larvae, reduced pupal weights significantly, and severely inhibited adult emergence. Larvae developing on ascorbic acid-free diet experienced far higher levels of mortality following per os infection with virus. Additionally, viral infection in larvae fed an ascorbic acid-free diet, as monitored by epifluorescence microscopy, showed signs of infection much earlier than larvae fed control levels of ascorbic acid. These results demonstrate an indirect correlation between the level of ascorbic acid in the food stream of larval H. virescens and the susceptibility of the insect to baculoviral infection.     

Ammonia toxicity as a criterion for the evaluation of larval quality in the prawn Macrobrachium rosenbergii

The feasibility of a short-term ammonia toxicity test as an evaluation criterion for larval quality was assessed in three trials. In each one, Macrobrachium rosenbergii larvae originating from the same spawn were nutritionally differentiated in two groups by feeding them either a nutrient-rich (Artemia nauplii enriched for 24 h with n-3 highly unsaturated fatty acids (HUFA) and ascorbic acid (AA)) or a nutrient-poor diet (Artemia nauplii starved for 24 h). Throughout their development, larvae from both treatments were exposed during 24 h to six concentrations of total ammonia (NH₄⁺+NH₃) and a control (no ammonia added). Based on mortality rates, the median lethal concentration for 50% of the population (LC₅₀) was estimated. As expected from earlier work, larvae fed the optimal diet presented higher n-3 HUFA and AA contents as well as higher growth and metamorphosis rates. From the moment the effect of diet quality was analytically detectable in the tissues of the larvae, the ammonia test was able to distinguish both groups of larvae. Differences in ammonia tolerance were observed as early as larval stage 4 and remained evident throughout larval development. The short-term ammonia toxicity test proved to be a valuable, sensitive and reproducible criterion for the establishment of larval quality.     

Ingestion rates and dietary lipids affect growth and fatty acid composition of Dendraster excentricus larvae

This study investigated the effect of single and mixed algal diets on growth, fatty acid composition and ingestion rates for Dendraster excentricus larvae. Larvae were assigned to three single algal diet treatments Isochrysis galbana, Dunaliella tertiolecta or Rhodomonas sp. and four mixed algal diet treatments D. tertiolecta and Rhodomonas, I. galbana and D. tertiolecta, I. galbana and Rhodomonas, D. tertiolecta, Isochrysis galbana and Rhodomonas sp. Small amounts (0.36-0.6%) of stearic acid (18:0) were seen in the three algae used but a relatively large percentage (7-25%) of this SAFA was found in Dendraster larvae. The alga D. tertiolecta had the highest percentage (51.7%) of the short chain polyunsaturated fatty acid (PUFA) linolenic acid 18:3(n-3) and trace amounts (0.02-0.14%) of the long chain PUFAs eicosapentanoic (EPA, 20:5(n-3) and docosahexanoic acids (DHA, 22:6(n-3)). However, sand dollar larvae demonstrated the ability to elongate and desaturate shorter chain (18 carbon) polyunsaturated fatty acids (PUFA) to longer chain (20 carbon) n-3 PUFA. Thus high levels of 18:3(n-3) in D. tertiolecta led to high levels of EPA and low levels of 18:3(n-3) in Dendraster larvae fed this diet. Rhodomonas sp. had the highest percentage of stearidonic acid (18:4(n-3), 38.14%) and EPA (10.6%). Despite high levels of 18:4(n-3) in Rhodomonas sp. this acid was absent or found at very low levels in larvae fed this alga, or any combination of this alga. I. galbana had the highest percentage of DHA(14.3%) but was almost devoid of EPA (0.43%). Although Rhodomonas sp. and I. galbana had high levels of EPA and DHA sand dollar larvae did not incorporate higher levels of these long chain PUFAs into their lipids compared to those fed the alga D. tertiolecta. Dendraster larvae synthesized a number of 20 and 22 carbon non-methylene interrupted dienes (NMID), with levels increasing with larval stage. Higher ingestion rates were observed for Dendraster larvae fed single algal diets (Rhodomonas sp. or D. tertiolecta) and lower ingestion rates for those fed mixed algal diets. The highest ingestion rates were for 8-arm Dendraster larvae fed the large alga Rhodomonas sp. presented as a single algal diet. When fed a combination of three algae, selection of particles varied slightly depending on stage with 8-arm larvae ingesting slightly more of the larger algal cell in the mixture than 6-arm larvae. The present study suggests that regardless of the ratios given larvae might have an optimum ratio of different sized particles at which they can feed. The mixed algal diet of I. galbana and D. tertiolecta was the best algal diet leading to significantly larger larvae with high survival and development to metamorphosis. The single algal diets of Rhodomonas sp. or Dunaliella tertiolecta were the second and third best algal diets based on growth and survival to metamorphosis.     

Plasma Ascorbic Acid,A Priori Diet Quality Score,and Incident Hypertension: A Prospective Cohort Study

Vitamin C may reduce risk of hypertension, either in itself or by marking a healthy diet pattern. We assessed whether plasma ascorbic acid and the a priori diet quality score relate to incident hypertension and whether they explain each other’s predictive abilities. Data were from 2884 black and white adults (43% black, mean age 35 years) initially hypertension-free in the Coronary Artery Risk Development in Young Adults Study (study year 10, 1995–1996). Plasma ascorbic acid was assessed at year 10 and the diet quality score at year 7. Eight-hundred-and-forty cases of hypertension were documented between years 10 and 25. After multiple adjustments, each 12-point (1 SD) higher diet quality score at year 7 related to mean 3.7 μmol/L (95% CI 2.9 to 4.6) higher plasma ascorbic acid at year 10. In separate multiple-adjusted Cox regression models, the hazard ratio of hypertension per 19.6-μmol/L (1 SD) higher ascorbic acid was 0.85 (95% CI 0.79–0.92) and per 12-points higher diet score 0.86 (95% CI 0.79–0.94). These hazard ratios changed little with mutual adjustment of ascorbic acid and diet quality score for each other, or when adjusted for anthropometric variables, diabetes, and systolic blood pressure at year 10. Intake of dietary vitamin C and several food groups high in vitamin C content were inversely related to hypertension, whereas supplemental vitamin C was not. In conclusion, plasma ascorbic acid and the a priori diet quality score independently predict hypertension. This suggests that hypertension risk is reduced by improving overall diet quality and/or vitamin C status. The inverse association seen for dietary but not for supplemental vitamin C suggests that vitamin C status is preferably improved by eating foods rich in vitamin C, in addition to not smoking and other dietary habits that prevent ascorbic acid from depletion.     

Effect of varying levels of dietary essential fatty acid during early ontogeny of the sea scallop Placopecten magellanicus

This study investigated the biological effects of various dietary essential fatty acids levels to sea scallop larvae, Placopecten magellanicus. Scallop larvae were fed three diets from D-veliger to settlement. Diet A consisted of Isochrysis sp. and Pavlova lutheri, diet B was a mix of Isochrysis sp. and Chaetoceros muelleri and diet C consisted of the same two species, but C. muelleri was grown under silicate deprivation to alter the fatty acid composition. Pediveligers (28 days old) were sampled prior to settlement for fatty acid analysis, growth measurement and survival assessment. Survival and settlement success were measured at the end of the experiment (day 40). Our results show that feeding regime greatly influenced larval size, settlement and fatty acid composition. Diet A was severely deficient in arachidonic acid (20:4n-6, AA), leading to the poorest larval growth, survival and lipid content. Nevertheless, larvae fed diet A selectively accumulate AA by a factor three compared to the dietary amount. Shell size of 28-day-old larvae was positively correlated with AA content and negatively correlated with eicosapentaenoic acid (20:5n-3, EPA)-AA ratio, thus suggesting that these two variables are of major interest for the optimisation of larval growth in sea scallops. Finally, larvae fed diet C displayed 20% higher shell size at day 28 than larvae fed diet A and B, likely in relation to the dietary amount of saturated fatty acid (SFA). However, the moderate survival and settlement success of these groups of larvae might be associated with a relative deficiency in docosahexaenoic acid (22:6n-3, DHA). This study underlines that the overall balance between polyunsaturated fatty acid (PUFA) needs to be considered to adequately fed sea scallop larvae.     

Utilization of fatty acids by Pieris brassicae reared on artificial and natural diets

The fatty acid composition of Pieris brassicae was measured from larvae reared on four different diets. Pieris can alter the composition of fatty acids in the diet through selective incorporation and synthesis. Oleate is preferentially accumulated on artificial diets (15·9 per cent in diet, 43·8 per cent in neutral lipid (NL) of fifth instar larvae), but not equally on natural diets (18·1 per cent in Brassica napus, 25·6 per cent in the NL of fifth instar larvae). Incorporation of linolenate appears to depend on the concentration of both linolenate and linoleate in the diet. With dietary levels of 35·7% linolenate and 32·2% linoleate, fifth instar larvae contain 12·2 and 16·0 per cent, respectively, of these acids. With 45·8% linolenate and 12·5% linoleate in the diet, fifth instar larvae contain 44·1 and 11·6 per cent of these acids, respectively, in the NL. Palmitoleate is actively synthetized on the artificial diets; with trace amounts of dietary palmitoleate, fifth instar larvae have 9·3 per cent of this acid in the NL. Pieris regulates the uptake of linoleate from the diet at the intestinal wall as was shown by linoleic acid-1-¹⁴C, and is unable to convert dietary linoleate to any of the 18-carbon analogues. The female apparently accumulates linolenate into egg phospholipids on the artificial diet, but in general the fatty acid composition of the eggs resembles that of the fat body.     

Effect of dietary polyunsaturated fatty acids on reproductive output and larval growth of bivalves

The pre-spawning condition of adult bivalves is influenced by quantity and quality of available food. For bivalves, the essential polyunsaturated fatty acids (PUFAs) eicosapentaenoic acid (EPA) 20:5(n-3) and docosahexaenoic acid (DHA) 22:6(n-3) are presumed to determine the nutritional value of algae. Macoma balthica kept on a broodstock diet supplemented with PUFAs spawned a larger number of eggs (average 22220) per female and larger sized eggs (106.7 μm) compared to adults kept on a diet without PUFA supplementation (962.5 eggs with average size 99.8 μm).Larvae of M. balthica from the same parental pool however did not profit from a diet where a part was replaced with PUFA spheres. Instead, larvae reared on Isochrysis sp. showed lower mortality and higher growth rates than larvae fed on the same algae supplemented with lipid spheres. Crassostrea gigas larvae showed no clear response to a PUFA supplemented diet.     

Vitamin requirements of larvae of Sitophilus oryzae

Improvements were made in the meridic larval diet for Sitophilus oryzae by replacing the minerals and vitamins supplied by dietary brewer's yeast and wheat germ with mineral and vitamin mixtures. The effects of different concentrations of individual vitamins were studied with the improved diet containing 20% casein. In later tests the dietary casein was replaced with a mixture of 15 amino acids. The results indicated that these larvae, which contain an associated bacteria-like micro-organism, required thiamine, nicotinic acid, pyridoxine, folic acid, and biotin in the diet. No requirement for riboflavin or pantothenic acid could be demonstrated on either the 20% casein diet or the amino acid diet possibly due to contamination of the cornstarch with these two vitamins. In addition, the larvae did not require choline or inositol for the growth of one larval generation.The asymbiotic larvae of S. granarius failed to develop on the improved casein diet indirectly implicating the symbiotes in a nutritional rôle; however, the symbiotes present in S. oryzae apparently do not provide B vitamins. Larvae of S. oryzae failed to develop when the concentration of casein was reduced to 10% while growth was maintained with a 10% concentration of the amino acid mixture. Casein is not an optimal source of amino acids for this species.     

Evidence for the protective effect of ascorbic acid (vitamin C) in treatment with γ-rays and chromium (VI) oxide (CrO3) in somatic cells of Drosophila

Evidence is provided that ascorbic acid (vitamin C), when used as a pretreatment, protects against mutation/recombination induced by γ-rays and chromium (VI) oxide in mwh+/+flr³ larvae in the wing spot test in Drosophila     

Developmental response of <Emphasis Type="Italic">Euplectrus comstockii</Emphasis> to ascorbic acid in the diet of the larval host, <Emphasis Type="Italic">Heliothis virescens</Emphasis>

Recent developments in genetic engineering have paved the way for researchers to produce crops of high nutritional and yield value, in addition to being resistant to diseases and pests. Ascorbic acid content is one of the parameters researchers are trying to enhance in plants. This study investigated the effect of different levels of dietary ascorbic acid of a beneficial wasp, Euplectrus comstockii Howard (Hymenoptera: Eulophidae), by measuring life history parameters of the wasp when reared on lepidopteran larvae fed a basal diet containing low and high levels of ascorbic acid. Odds and odds ratio analyses showed that the probability of egg hatch and adult emergence for the wasp increased with the amount of ascorbic acid in the diet of the host, and that the rate of development and probability of female or male progeny was similar for most levels of ascorbic acid tested. This would indicate that as the ascorbic acid concentration increases in the pest insect the effectiveness of the wasp is likely to increase and when, by comparison with other published findings, the effectiveness of microbial pathogens is likely to decrease.     

Chromium and chronic ascorbic acid depletion effects on tissue ascorbate,manganese, and14C retention from14C-ascorbate in guinea pigs

Chromium (Cr) potentiates the effects of insulin and a role for insulin in ascorbic acid transport has been reported. Therefore, the effects of Cr and ascorbate depletion on tissue ascorbic acid and¹⁴C distribution and excretion after a¹⁴C ascorbate dose were investigated in guinea pigs. As utilization of dietary Cr is affected by interaction with other minerals, tissue manganese (Mn), zinc (Zn), copper (Cu), and iron (Fe) were examined. For 20 wk, 40 weanling animals were fed either a Cr-deficient (<0.06 μg Cr/g diet, ?Cr) or a Cr-adequate (2 μg Cr from CrCl₃/g diet, +Cr) casein-based diet and were given 1 mg ascorbate/d (?C) or 10 mg ascorbate/d (+C) for 20 wk. Animals fed the Cr-depleted diet had decreased weight at 20 wk (p<0.01). Six hours before necropsy, animals were dosed by micropipette with 1.8 μCi ofl-[carboxyl-¹⁴C] ascorbic acid and placed in metabolic cages. Ascorbate supplementation increased Fe concentrations in most analyzed tissues, hepatic¹⁴C, tissue ascorbate and Mn concentration in the adrenal and testes, but decreased the concentrations of Cu in the kidney and Mn in the spleen. Liver Mn concentration was higher and kidney Mn concentration was lower in +Cr animals. Interactions between Cr and ascorbic acid affected Mn concentrations in bone and brain. These results indicate that ascorbate and Cr may affect Mn distribution. Chromium supplementation decreased plasma cortisol, brain¹⁴C and the amount of¹⁴C expired as carbon dioxide. These findings suggest that dietary Cr may affect ascorbic acid metabolism and the metabolic response to stress.     

Lipid content and fatty acid composition of larvae and adults of the velvetbean caterpillar, Anticarsia gemmatalis, as affected by larval diet

Eight fatty acids were found in the food, larvae and adults of the velvetbean caterpillar, Anticarsia gemmatalis, with C16:0, C18:0, C18:1, C18:2 and C18:3 accounting for over 90% of the total. Fatty acid composition of the larvae tended to reflect that of their food. The most striking differences were the high percentages of C18:1 and C18:2 and the low percentage of C18:3 in the artificial diet and artificial diet-reared larvae compared to the foliage of three species of legume food plants (soybean, Glycine max; pigeon pea, Cajanus cajan; and hairy indigo, Indigofera hirsuta) and foliage-reared larvae. Lipid content (%dw) declined during metamorphosis from mature larva to adult with diet-reared larvae and adults exhibiting significantly higher lipid contents than foliage-reared insects. Regardless of larval food, newly eclosed adults tended to exhibit a decrease in the % of C18:3, and increases in C16:0 and C18:1 compared to larvae. Larval diet clearly influenced the fatty acid composition of larvae and adults, but only the % of C18:2 did not change between food, larvae and adults.     

Effects of the levels of fatty acids and carbohydrates in a diet on the biosynthesis of fatty acids in larvae of the silkworm, Bombyx mori

The effects of varying levels of fatty acids and carbohydrates in the diet on fatty acid synthesis from glucose in the larvae of the silkworm, Bombyx mori, were investigated. Elevation of the level of dietary fatty acids resulted in the decrease of the rate of fatty acid synthesis in the larvae. The addition of palmitate, stearate, or oleate to a diet had an inhibitory effect on fatty acid synthesis. The prolonged feeding of larvae on a diet containing a high level of fatty acid intensified the depression of the synthesis. The inhibitory effect of dietary fatty acid was found in the presence of both high and low levels of dietary carbohydrates. On the other hand, the rate of fatty acid synthesis was greatly accelerated by increasing the level of sucrose in a diet but not by the addition of starch. Furthermore, the fatty acid composition of the larval tissue shows a marked difference between the two groups of larvae fed on a diet containing sucrose and on a diet containing potato starch. Palmitic and oleic acid contents of larval tissue were increased significantly on the sucrose diet.     

Inhibition of reproduction of Xyleborus ferrugineus by ascorbic acid and related chemicals

Effects of various dietary chemicals on the reproduction of the ambrosia beetle, Xyleborus ferrugineus were studied. Ascorbic acid, araboascorbic acid, dehydroascorbic acid, hydroquinone, catechol, cysteine, and α-tocopherol each inhibited progeny production by virgin females. Detailed studies of the effects of ascorbic acid showed that it inhibited progeny production by causing a nutritionally detrimental non-enzymic browning of the dietary casein. Amino acid analyses of such browned and unbrowned casein, after in vitro digestion with pepsin and pancreatin, showed that lesser amounts of certain amino acids were released from the browned material. Effects of the non-enzymic browning were overcome by increasing the casein component of the diet. It was further concluded that araboascorbic acid, dehydroascorbic acid, hydroquinone, and catechol probably inhibit reproduction of X. ferrugineus by the same mechanism as ascorbic acid. No explanation of the inhibitory action of α-tocopherol on X. ferrugineus is offered.     

A freeze-dried diet to test pathogens of Colorado potato beetle

The Colorado potato beetle is an important pest on potato, eggplant, and tomato. Because Colorado potato beetles develop resistance to insecticides quickly, new methods are needed for control. Bacillus thuringiensis is the only bacterium to successfully control Colorado potato beetle. Until recently, one of the drawbacks to testing bacteria against the Colorado potato beetle has been the lack of an artificial diet for screening. Previous artificial diets will only be consumed by Colorado potato beetle larvae when fresh. To improve storage, we developed a freeze-dried diet, based on a 96-well plate, suitable to feed larvae for the duration of a bioassay. Individual diet components were tested both for their effect on insect growth and on pathogen toxicity. When the preservatives, methylparaben and sorbic acid, were removed from the diet, the average weight of second instar larvae increased from 7.9 mg to greater than 9.8 mg. The preservatives inhibited the growth of two of the bacteria tested, Photorhabdus luminescens HM and Chromobacterium sp. PRAA. The removal of these preservatives also allowed for fungal growth and reduced survival from 94 to 38%. Removing diet preservatives, that inhibited the growth of Chromobacterium sp. PRAA, increased the total mortality of the larvae as well as reducing the time needed to kill 50% of the larvae. Compared to incorporation of bacteria into molten diet, the total mortality of Colorado potato beetle fed either P. luminescens HM or Chromobacterium sp. PRAA on freeze-dried diet doubled. Preparation of freeze-dried diet need not be synchronized with the insect or the pathogen. The freeze-dried diet gave consistent results as measured by low control mortality and pathogen toxicity over time.     

Methanol metabolism in the Asian corn borer, Ostrinia furnacalis (Guenée) (Lepidoptera: Pyralidae)

Plants produce and release large quantities of methanol, especially when attacked by herbivores. It seems that the herbivores may suffer from methanol intoxication. Here we reported the tolerance to and the metabolism of methanol by Ostrinia furnacalis third-instar larvae. When larvae were exposed to dietary methanol, formaldehyde and formic acid for 72 h, the estimated LC₅₀ value was 28, 40 and 29 mg/g diet, respectively. Toxicity of methanol was enhanced by 4-methylpyrazole, 3-amino-1,2,4-triazole and piperonyl butoxide, and toxicity of formaldehyde was increased by 3-amino-1,2,4-triazole and piperonyl butoxide. However, triphenyl phosphate had little synergistic effects on both methanol and formaldehyde. These data indicate that alcohol dehydrogenase, and probably catalase and cytochrome P450 monooxygenase oxidize methanol to formaldehyde, catalase and cytochrome P450 monooxygenase catalyze formaldehyde to formic acid, water and carbon dioxide, and carboxylesterase may have a minor effect. Several fatty acid methyl esters (FAMEs) were identified from extracts of the frass of larvae which had been exposed to a methanol-contained diet, in contrast to those on a methanol-free artificial diet. In vitro tests revealed that a crude enzyme solution from the larvae could synthesize FAMEs from corresponding fatty acids and methanol. In addition, dietary methanol induced higher esterase activities in the first-, second- and third-instar larvae. These findings demonstrate that both oxidative metabolism and non-oxidative metabolism are partially responsible for methanol elimination in O. furnacalis larvae.     

Cool-textured diets for use in baculovirus production

The incorporation of baculovirus into semi-synthetic diets has been used for the inoculation of large batches of insect larvae reared under gregarious conditions. The preparation of most artificial diets involves heating to dissolve the agar gelling agent followed by cooling to ～55–60°C whereupon virus occlusion bodies (OBs) are incorporated into the diet. To avoid exposure of thermolabile OBs to high temperatures, a cool-textured diet using soy fiber was evaluated as a simple method to inoculate batches of larvae with the baculovirus OBs. Spodoptera exigua larvae that fed on a diet with 120, 140 or 160?g/l of soy fiber, as a substitute for agar, had a similar weight gain over a 24?h period than larvae that consumed a standard agar diet. The larval weight gain with cool-textured diets with 140?g/l of soy fiber was not improved by the addition of chlorophyll, green food dye colour, or by the removal of ascorbic acid, sodium benzoate and potassium sorbate. Preliminary tests performed to determine the optimal OBs concentration needed for the inoculation of groups of four instar larvae in agar and soy fiber diets revealed no virus inactivation in heated diets with a similar optimal concentration of 36?OBs?mg^?1 for both diets. Moreover, no significant differences were observed in OB production when each type of diet was used to inoculate groups of 250 larvae reared gregariously in plastic containers. Cool-textured diets could be of interest for the production of baculoviruses that require a diet that can be prepared quickly and easily.     

Comparative food utilization of coffee bean weevil larvae on a new freeze-dried diet and green coffee beans

A meridic diet for culturing the coffee bean weevil, Araecerus fasciculatus was formulated and freeze-dried into pellets. When exposed to an atmosphere of 80 ± 5%r.h. the diet reached a moisture equilibrium that invoked oviposition and was satisfactory for growth and development of the insect. Larvae ingested larger amounts of the freeze-dried diet than of the natural food, green coffee beans, reaching equivalent weight and pupation in 31.8 ± 0.4 days on the diet as compared to 46.8 ± 0.8 days on the coffee beans. Approximate digestibilities of the two foods were not significantly different. However, the coffee beans were converted more efficiently to body mass than was the diet. Faeces of larvae on the diet contained more uric acid than those of larvae on coffee beans. Continuous culture and mass rearing of A. fasciculatus on the diet is possible.     

Silencing of acetylcholinesterase gene of Helicoverpa armigera by siRNA affects larval growth and its life cycle

RNA interference is an effective means of regulation of gene expression both in vitro and in vivo. We studied the effect of siRNA on larval development by selective targeting of the acetylcholinesterase (AChE) gene of Helicoverpa armigera. Chemically synthesized siRNA molecules were directly fed to H. armigera larvae along with the artificial diet. The siRNA treatment resulted in specific gene silencing of AChE and consequently brought about mortality, growth inhibition of larvae, reduction in the pupal weight, malformation and drastically reduced fecundity as compared to control larvae. Our studies suggest some novel roles for AChE in growth and development of insect larvae and demonstrate that siRNA can be readily taken up by insect larvae with their diet.     

Ascorbic acid co-administration with a low dose of diminazene aceturate inhibits the in vitro growth of Theileria equi,and the in vivo growth of Babesia microti

In the current investigation, the effect of ascorbic acid on the in vitro growth of several piroplasm including Babesia bovis (Bartonella bovis), Baconia bigemina, B. caballi, and Theileria equi (T. equi), as well as against Brucella microti in mice was assessed. The antipiroplasm efficacy of ascorbic acid in vitro and in vivo was assessed using a fluorescence-based SYBR Green I test. Using atom pair fingerprint (APfp), we investigated the structural similarity between ascorbic acid and the commonly used antibabesial medicines, diminazene aceturate (DA) and imidocarb dipropionate (ID). In vitro cultures of B. bovis and T. equi were utilized to determine the ascorbic acid and DA interaction using the Chou–Talalay method. Ascorbic acid inhibited B. bovis, B. bigemina, T. equi, and B. caballi growth in vitro in a dose-dependent manner. The APfp results revealed that ascorbic acid and DA have a maximum structural similarity (MSS). On a T. equi culture in vitro, ascorbic acid showed a synergistic interaction with DA, with a combination index of 0.28. B. microti growth was decreased by 41% in vivo using ascorbic acid combined with a very low dosage of DA (6.25 mg kg⁻¹). The results imply that ascorbic acid /DA could be a viable combination therapy for the treatment of T. equi and that it could be utilized to overcome the resistance of Babesia parasites to full doses of the regularly used antibabesial medication, DA.