Development and Evaluation of a Stable Oil-in-Water Emulsion with High Ostrich Oil Concentration for Skincare Applications.

This study investigates the development of an oil-in-water (O/W) emulsion enriched with a high concentration of ostrich oil, recognized for its abundant content of oleic acid (34.60 ± 0.01%), tailored for skincare applications. Using Span and Tween emulsifiers, we formulated an optimized emulsion with 20% w/w ostrich oil and a 15% w/w blend of Span 20 and Tween 80. This formulation, achieved via homogenization at 3800 rpm for 5 min, yielded the smallest droplet size (5.01 ± 0.43 µm) alongside an appropriate zeta potential (-32.22 mV). Our investigation into the influence of Span and Tween concentrations, types, and ratios on the stability of 20% w/w ostrich oil emulsions, maintaining a hydrophile-lipophile balance (HLB) of 5.5, consistently demonstrated the superior stability of the optimized emulsion across various formulations. Cytotoxicity assessments on human dermal fibroblasts affirmed the safety of the emulsion. Notably, the emulsion exhibited a 52.20 ± 2.01% inhibition of linoleic acid oxidation, surpassing the 44.70 ± 1.94% inhibition observed for ostrich oil alone. Moreover, it demonstrated a superior inhibitory zone against Staphylococcus aureus (12.32 ± 0.19 mm), compared to the 6.12 ± 0.15 mm observed for ostrich oil alone, highlighting its enhanced antioxidant and antibacterial properties and strengthening its potential for skincare applications. The optimized emulsion also demonstrates the release of 78.16 ± 1.22% of oleic acid across the cellulose acetate membrane after 180 min of study time. This successful release of oleic acid further enhances the overall efficacy and versatility of the optimized emulsion. Stability assessments, conducted over 6 months at different temperatures (4 °C, 25 °C, 45 °C), confirmed the emulsion's sustained physicochemical and microbial stability, supporting its promise for topical applications. Despite minor fluctuations in acid values (AV) and peroxide values (PV), the results remained within the acceptable limits. This research elucidates the crucial role of emulsification in optimizing the efficacy and stability of ostrich oil in skincare formulations, providing valuable insights for practical applications where stability is paramount.

The first record of ostrich feather louse (Struthiolipeurus struthionis) collected from farmed ostriches (Struthio camelus) in the United Arab Emirates.

Background and Aim: Ostrich (Struthio camelus) farming in the United Arab Emirates (UAE) is a relatively new field of farming. Farmed ostriches are susceptible to ectoparasite infestation, which affects their production. This study was conducted to estimate the prevalence of ectoparasites on ostriches raised on a farm in Abu Dhabi Emirate. Materials and Methods: The feathers of 42 ostriches (26 females and 16 males) were collected and morphologically examined for ectoparasites. In total, 283 lice (89 males and 194 females) were collected from birds. However, there were no ticks or other ectoparasites. Lice were preserved in 1.5 mL tubes containing 70% ethanol and were later identified using taxonomic keys. The prevalence, mean intensity of infection, and mean abundance were estimated. Results: One louse species, Struthiolipeurus struthionis was identified. To the best of our knowledge, this is the first report of S. struthionis in ostriches raised in the UAE with an overall prevalence of 88%. The prevalence of lice was significantly higher in July (100%) than that in May (66.7%). Likewise, the mean intensity and abundance of lice were significantly higher in June (10.6 and 9.94, respectively) than in May (3.25 and 2.2, respectively). Conclusion: The high prevalence of lice poses a serious risk to ostrich farming by negatively affecting the health and productivity of ostriches.

Anatomical and Functional Study of the Ostrich (Struthio camelus) Lung through Macroscopic Analysis in Combination with Optical and Electron Microscopy Techniques.

The Ostrich occupies a unique position as the largest bird on the planet. Like other ratites, it has been reputed to have a phylogenetically primitive lung. We used macroscopy, light microscopy, transmission and scanning electron microscopy as well as silicon rubber casting to elucidate the functional design of its lung and compare it with what is already documented for the avian species. The neopulmonic region was very small and poorly developed. The categories of the secondary bronchi (SB) present and their respective numbers included laterodorsal (8-10), lateroventral (4-5), medioventral (4-6) and posterior (16-24). The lateral aspects of the laterodorsals were covered with a transparent collapsible membrane internally lined with a squamous to cuboidal epithelium. The bulk of these SB were in close proximity to intercostal spaces and the intercostal muscles and were thought to be important in the propulsion of gases. The lung parenchyma was rigid, with the atria well supported by septa containing smooth muscles, connective tissue interparabronchial septa were absent, and blood capillaries were supported by epithelial bridges. There were two categories of epithelia bridges: the homogenous squamous type comprising two leaflets of type I cells and the heterogeneous type consisting of a type I pneumocyte and type II cell. Additional type two cells were found at the atrial openings as well as the walls of the infundibulae and the air capillaries. The atria were shallow and opened either directly into several air capillaries or into a few infundibulae. The presence of numerous type II cells and the absence of interparabronchial connective tissue septa may imply that the ostrich lung could be capable of some degree of compliance.

Ostrich (Struthio camelus) syrinx morphology and vocal repertoire across postnatal ontogeny and sex: Implications for understanding vocal evolution in birds.

Vocal production in birds has been the target of considerable research that mostly has focused on phylogenetically well-nested songbirds. Anatomical descriptions and recordings of many non-songbirds have often only focused on a single ontogenetic stage or sex. While basic morphology of the vocal organ (syrinx) of ostrich (Palaeognathae, Struthio camelus) has been known since the 1800s, descriptions of its vocal repertoire and syrinx anatomy since then have been incomplete or inconsistent. New toolkits now enable detailed qualitative description of internal anatomy and meristic data and allow it to be compared to vocal production. Here we describe the anatomy of the syrinx in Struthio camelus for three post-hatching ontogenetic stages and both an adult male and female utilizing dissection and contrast enhanced X-ray computed tomography (diceCT). We find changes in ring geometry and spacing through ontogeny as well as lateral labia thickness. We document a small unpaired, midline, cartilaginous structure, a "pessuliform process" at the tracheobronchial juncture present throughout ontogeny and in both males and females. Investigation of the vocal repertoire of ostriches across ontogeny using a new dataset of 77 recordings led to identification of four vocalizations not previously reported in the literature, including the simultaneous production of a hiss and tonal. We find syrinx morphology largely consistent across ontogeny and in male and female adults. Both are capable of producing long duration tonal calls, but these may be more frequent in male birds. Closed-mouth boom calls remain unique to males. A detailed understanding of diversity in parts of early diverging clades is pivotal in attempting to estimate features of the ancestral syrinx in birds and how avian vocalization evolved.

Information Avoidance: Past Perspectives and Future Directions.

In the present age of unprecedented access to information, it is important to understand how and why people avoid information. Multiple definitions of "information avoidance" exist, and key aspects of these definitions deserve attention, such as distinguishing information avoidance from (lack of) information seeking, considering the intentionality and temporal nature of information avoidance, and considering the personal relevance of the information. In this review, we provide a cross-disciplinary historical account of theories and empirical research on information avoidance and seeking, drawing from research in multiple fields. We provide a framework of antecedents of information avoidance, categorized into beliefs about the information (e.g., risk perceptions), beliefs about oneself (e.g., coping resources), and social and situational factors (e.g., social norms), noting that constructs across categories overlap and are intertwined. We suggest that research is needed on both positive and negative consequences of information avoidance and on interventions to reduce information avoidance (when appropriate). Research is also needed to better understand temporal dynamics of information avoidance and how it manifests in everyday life. Finally, comprehensive theoretical models are needed that differentiate avoidance from seeking. Research on information avoidance is quickly expanding, and the topic will only grow in importance.

Construction of Chicken and Ostrich Antibody Libraries.

Recombinant antibody libraries based on chicken immunoglobulin genes are potentially valuable sources of phage-displayed scFvs for use in veterinary diagnostics and research. To add diversity to the scFv repertoire, we expanded the library to include genes from the ostrich, indigenous to southern Africa. The libraries described in this chapter are based on the chicken and ostrich variable heavy and light chain immunoglobulin genes joined by a short flexible linker cloned in the phagemid vector pHEN1. The resulting phagemids produce either scFvs displayed on the surface of the fusion phage subsequent to co-infection with helper phage or soluble scFvs following IPTG induction. This chapter provides detailed and proven methods for the construction of such libraries.

Partial Amputation of a Nonreducing Prolapsed Phallus in an Ostrich (Struthio camelus).

A 19-year-old male ostrich (Struthio camelus) was referred to a veterinary teaching hospital (São Paulo State University, Botucatu, Brazil) due to a 6-month history of recurrent prolapse of the phallus. On physical examination, 2 ulcerative wounds were present on the phallus, as well as caseous plaques and myiasis. Conservative treatment resulted in improvement but prolapse of the phallus remained. Thus, a decision was made to perform a partial phallectomy. The surgery was successful and no postoperative complications occurred. When the ostrich was reexamined 6 months postsurgery, the ostrich was alert, in good health, and the surgical site completely healed. The owner verbally reported no recurrence of the phallus prolapse 1 year after surgery.

Morphometric studies on the appendicular bony skeleton of the ostriches (Struthio Camelus).

BACKGROUND: Morphometric study of the bony elements of the appendicular skeleton in the ostrich was fully described and identified. The appendicular skeleton included the bones of the pectoral girdle, the wing, the pelvic girdle and the pelvic limb. RESULTS: The shoulder girdle of the ostrich included the scapula and coracoid bones. The scapula appeared as a flattened spoon-like structure. The coracoid bone appeared quadrilateral in outline. The mean length of the scapula and coracoid (sternal wing) were 15.00 ± 0.23 and 10.00 ± 0.17 cm, respectively. The wing included the humerus, ulna, radius, radial carpal bone, ulnar carpal bone, carpometacarpus and phalanges of three digits. The mean length of the humerus, radius, and ulna were 33.00 ± 0.46, 10.50 ± 0.40 and 11.50 ± 0.29 cm respectively. The carpometacarpus was formed by the fusion of the distal row of carpal bones and three metacarpal bones. Digits of the wing were three in number; the alular, major and minor digits. Os coxae comprised the ilium, ischium and pubis. Their mean lengths were 36.00 ± 0.82 cm, 32.00 ± 0.20 and 55.00 ± 0.2.9 cm, respectively. The femur was a stout short bone, that appeared shorter than the tibiotarsus. The mean length of the femur, tibiotarsus, and tarsometatarsus were 30.00 ± 0.23, 52.00 ± 0.50 and 46.00 ± 0.28 cm. Tibiotarsus was the longest bone in the pelvic limb. The fibula was a long bone (44.00 ± 0.41 cm) lying along the lateral surface of the tibiotarsus. The tarsometatarsus was a strong long bone formed by the fusion of the metatarsal (II, III, IV) and the distal row of tarsal bones. It was worth mentioning that metatarsal II was externally absent in adults. CONCLUSIONS: In the appendicular skeleton of ostrich, there were special characteristic features that were detected in our study; the clavicle was absent, the coracoid bone was composed of a sternal wing and scapular wing, the ulna was slightly longer in length than the radius. The coupled patellae i.e., the proximal and distal patella were observed; and the ostrich pedal digits were only two; viz., the third (III) and fourth (IV) digits.

Steps of spermiogenesis in the ostrich (Struthio camelus).

Few studies describe the sequence of morphological events that characterize spermiogenesis in birds. In this paper, the clearly observable steps of spermiogenesis are described and illustrated for the first time in a commercially important ratite, the ostrich, based on light microscopy of toluidine blue-stained plastic sections. Findings were supplemented and supported by ultrastructural observations, PNA labeling of acrosome development, and immunocytochemical labeling of isolated spermatogenic cells. Spermiogenesis in the ostrich followed the general pattern described in non-passerine birds. Eight steps were identified based on changes in nuclear shape and contents, positioning of the centriolar complex, and acrosome development. Only two steps could be recognized with certainty during development of the round spermatid which contributed to the fewer steps recorded for the ostrich compared to that described in some other bird species. The only lectin that displayed acrosome reactivity was PNA and only for the first three steps of spermiogenesis. This suggests that organizational and/or compositional changes may occur in the acrosome during development and merits further investigation. Immunological labeling provided additional evidence to support the finding of previous studies that the tip of the nucleus in the ostrich is shaped by the forming acrosome and not by the microtubular manchette. To our knowledge, this is the first complete description of spermiogenesis in ostrich and one of few in any avian species. In addition to comparative reproduction and animal science, this work has implications for evolutionary biology as the reported germ cell features provide a bridge between reptile and ratite-avian spermatogenesis.

Methods of In Ovo and Ex Ovo Ostrich Embryo Culture with Observations on the Development and Maturation of the Chorioallantoic Membrane.

Culture of shell-free and windowed eggs for drug testing and other experiments has been perfected for smaller eggs such as those of chickens, where the developing blood vessels of the chorioallantoic membrane (CAM) become accessible for manipulative studies. However, due to the thickness and hardness of the ostrich egg shell, such techniques are not applicable. Using a tork craft mini rotary and a drill bit, we established windowed egg, in-shell-membrane windowed egg, and in-shell-membrane shell-free methods in the ostrich egg, depending on whether the shell membranes were retained or not. Concomitant study of the developing CAM revealed that at embryonic day 16 (E16), the three layers of the CAM were clearly delineated and at E25, the chorionic capillaries had fused with the epithelium while the CAM at E37 had reached maturity and the chorion and the allantois were both 3-4 times thicker and villous cavity (VC) and capillary-covering cells were well delineated. Both intussusceptive and sprouting angiogenesis were found to be the predominant modes of vascular growth in the ostrich CAM. Development and maturation of the ostrich CAM are similar to those of the well-studied chicken egg, albeit its incubation time being twice in duration.

Coprophagy rapidly matures juvenile gut microbiota in a precocial bird.

Coprophagy is a behavior where animals consume feces, and has been observed across a wide range of species, including birds and mammals. The phenomenon is particularly prevalent in juveniles, but the reasons for this remain unclear. One hypothesis is that coprophagy enables offspring to acquire beneficial gut microbes that aid development. However, despite the potential importance of this behavior, studies investigating the effects in juveniles are rare. Here we experimentally test this idea by examining how ingestion of adult feces by ostrich chicks affects their gut microbiota development, growth, feeding behavior, pathogen abundance, and mortality. We conducted extensive longitudinal experiments for 8 weeks, repeated over 2 years. It involved 240 chicks, of which 128 were provided daily access to fresh fecal material from adults and 112 were simultaneously given a control treatment. Repeated measures, behavioral observations, and DNA metabarcoding of the microbial gut community, both prior to and over the course of the experiment, allowed us to evaluate multiple aspects of the behavior. The results show that coprophagy causes (a) marked shifts to the juvenile gut microbiota, including a major increase in diversity and rapid maturation of the microbial composition, (b) higher growth rates (fecal-supplemented chicks became 9.4% heavier at 8 weeks old), (c) changes to overall feeding behavior but no differences in feed intake, (d) lower abundance of a common gut pathogen (Clostridium colinum), and (e) lower mortality associated with gut disease. Together, our results suggest that the behavior of coprophagy in juveniles is highly beneficial and may have evolved to accelerate the development of gut microbiota.

Antimicrobial activity and wound healing effect of a novel natural ointment: an in vitro and in vivo study.

Infection and pathological disorders, such as cellular disorders, ischaemia, neuropathy and angiogenesis, are considered the most critical factors which cause a delay in the wound healing process in patients with diabetes. This study aimed to investigate the effect of an ointment based on ostrich oil containing honey, beeswax, and ethanolic extracts of Nigella sativa, propolis and Cassia angustifolia on the wound healing process of diabetic rats. Gas chromatography/mass spectrometry analysis showed caffeic acid and pinostrobin chalcone molecules present in propolis, giving antibacterial and antifungal properties to the compound. The antibacterial assessment showed the ointment had remarkable antibacterial activity against Staphylococcus aureus (8.6±0.28mm), Escherichia coli (9.4±0.31mm), Acinetobacter baumannii (7.2±0.23mm) and Pseudomonas aeruginosa (13.9±0.42mm). In vivo results showed the ointment significantly accelerated wound healing and increased collagen deposition compared with the control (p<0.05). Histopathology evaluation also showed hair follicles, sebaceous glands and vessels in the group that used the ointment. These results proved successful and diabetic wound healing was rapid. Therefore, it could be concluded that the fabricated ointment could be a suitable candidate for wound healing.

Assessment of the microbiological status of two Hungarian ostrich farms.

The aim of this study was to estimate the occurrence of bacterial infection and contamination in two ostrich-producing farms. Compared to other poultry species, the hatchability of ostrich eggs is especially low. In a quest to identify factors that may affect hatchability, we collected faecal samples from adult birds, as well as eggs with dead-in-shell embryos, dead chicks and swab samples from the surface of the eggs and from the environment. The samples were screened for the presence of bacteria by routine bacteriological culture methods. The most prevalent bacteria, detected in the samples, were Escherichia coli, Bacillus spp. and coliform bacteria, whereas Pseudomonas spp. were less frequently found. The intensity and species compositon of the bacterial contamination was comparable in the two farms. Our results revealed that the bacteria, present in the environment, may likely be transmitted to the surface of the eggs. If they are able to penetrate the shell then the embryos and chicks become infected easily. These findings draw the attention to the special importance of enforcing efficient decontamination and disinfection measures to keep the environment and egg surface free from germs. Besides the appropriate egg treatment procedure, the incubation and hatching technology should also be kept under control.

A Novel Breast Cancer Xenograft Model Using the Ostrich Chorioallantoic Membrane-A Proof of Concept.

The avian chorioallantoic membrane (CAM) assay has attracted scientific attention in cancer research as an alternative or complementary method for in vivo animal models. Here, we present a xenograft model based on the ostrich (struthio camelus) CAM assay for the first time. The engraftment of 2 × 106 breast cancer carcinoma MDA-MB-231 cells successfully lead to tumor formation. Tumor growth monitoring was evaluated in eight fertilized eggs after xenotransplantation. Cancer cells were injected directly onto the CAM surface, close to a well-vascularized area. Histological analysis confirmed the epithelial origin of tumors. The CAM of ostrich embryos provides a large experimental surface for the xenograft, while the comparably long developmental period allows for a long experimental window for tumor growth and treatment. These advantages could make the ostrich CAM assay an attractive alternative to the well-established chick embryo model. Additionally, the large size of ostrich embryos compared to mice and rats could help overcome the limitations of small animal models. The suggested ostrich model is promising for future applications, for example, in radiopharmaceutical research, the size of the embryonal organs may compensate for the loss in image resolution caused by physical limitations in small animal positron emission tomography (PET) imaging.

Ostrich oil as a fat substitute in milk-based infant formula.

In this study, the possibility of replacing vegetable fats with ostrich oil in infant formula (IF) production was investigated. The fatty acid profile, the positional distribution of fatty acids in the triacylglycerols, the cholesterol content, and the physicochemical properties of ostrich oil were determined and compared with breast milk fat and vegetable oils. In the next step, two infant formulas were produced using ostrich oil and vegetable oils and the physicochemical properties, rheological properties, color parameters, and sensory analysis of the resultant powders were compared. The results showed that the predominant fatty acids in ostrich oil are palmitic acid, oleic acid, and linoleic acid which is similar to breast milk fat and vegetable oils. The presence of appropriate cholesterol content in ostrich oil makes it more similar to breast milk fat compared to vegetable fats. Palmitic acid was located at sn-2 position in 15% triacylglycerol from ostrich fat, which was equal to the amount reported for vegetable fats. The incorporation of ostrich oil in infant formula production showed that there is no statistically significant difference between quality attributes of powder formulated with ostrich oil or vegetable oils. Therefore, ostrich oil can be introduced as a new source of edible oil, and addition of ostrich oil is an effective way to reduce the gap between the composition of breast milk and infant formula.

Subtotal Cervical Esophagectomy and Anastomosis in an Ostrich (Struthio camelus domesticus).

An adult, female, captive ostrich (Struthio camelus domesticus) was referred to a veterinary teaching hospital for a 2-week history of lethargy and a mass effect in the proximal cervical region. Physical examination revealed a fistula in the middle cervical esophagus surrounded by devitalized and necrotic tissue; feed material was found leaking from the site. Cervical radiography identified an esophageal stricture with anterior dilation due to the accumulation of feed. After receiving supportive care for 48 hours, the patient's overall status improved, allowing partial esophagectomy and resection of the affected tissues with end-to-end anastomosis. Postoperative management included fasting for 24 hours, followed by the administration of a liquid hand-rearing formula prepared with commercially available ostrich feed and administered via a feeding tube for 15 days. Proper healing of the surgical site was confirmed by esophagoscopy using a flexible endoscope 17 days after surgery. The ostrich was discharged after 27 days, with no complications recorded within the 180 days of the follow-up period. Partial cervical esophagectomy with end-to-end anastomosis along with pre- and postoperative management provided a successful outcome for the treatment of a fistulated esophageal stricture in a captive ostrich, resulting in full recovery without surgical complications.

Smart Biomechanical Adaptation Revealed by the Structure of Ostrich Limb Bones.

Ostriches are known to be the fastest bipedal animal alive; to accomplish such an achievement, their anatomy evolved to sustain the stresses imposed by running at such velocities. Ostriches represent an excellent case study due to the fact that their locomotor kinematics have been extensively studied for their running capabilities. The shape and structure of ostrich bones are also known to be optimized to sustain the stresses imposed by the body mass and accelerations to which the bones are subjected during movements. This study focuses on the limb bones, investigating the structure of the bones as well as the material properties, and how both the structure and material evolved to maximise the performance while minimising the stresses applied to the bones themselves. The femoral shaft is hollowed and it presents an imbricate structure of fused bone ridges connected to the walls of the marrow cavity, while the tibial shaft is subdivided into regions having different mechanical characteristics. These adaptations indicate the optimization of both the structure and the material to bear the stresses. The regionalization of the material highlighted by the mechanical tests represents the capability of the bone to adapt to external stimuli during the life of an individual, optimizing not only the structure of the bone but the material itself.

Comparison of embryonic development, from HH21 to HH40, between ostrich (Struthio camelus) and chicken (Gallus gallus).

BACKGROUND: The chicken has been a representative model organism to study embryonic development in birds, however important differences exist among this class of species. As a representative of one of oldest existing clades of birds, the African ostrich (Struthio camelus), has the largest body among birds, and has two toes. Our purpose is to establish the corresponding stages in ostrich embryo development that match the well-established HH system of the chicken to facilitate comparative studies between the ostrich and other birds to better understand differences in development. RESULTS: Here we describe in detail the middle period of embryonic development using microscopic images and skeletal staining. We found that clear morphological differentiation between the ostrich and the chicken begins at stage 26. Bird limb cartilage first form in stage 25, while the development of the limb skeletons differs after stage 31. Calcification of limb skeletons in the chicken was completed faster. The first and second toes of the ostrich disappear at stages 36 and 38, respectively. CONCLUSIONS: This study should greatly aid ostrich-related developmental and morphological research and provide a reference for studying the development and evolution of avian limb skeletons, including molecular research. Questions that can now be addressed include studies into the fusion of tarsometatarsal skeleton, ossification, and digit loss.

Morphological and Transcriptomic Analysis of the Supplemental Boron in the Liver of Ostrich Chicks.

African ostrich chicks (Struthio camelus) were divided into six groups, and each received different levels of boric acid (source of boron) in the drinking water (0, 40, 80, 160, 320, and 640 mg/L respectively) to examine the histological, apoptotic, biochemical, and transcriptomic parameters. Morphological analysis in different groups was assessed by hematoxylin and eosin (H&E) staining, periodic acid Schiff (PAS) staining, and terminal deoxynucleotide transferase dUTP Nick-End Labeling (TUNEL) assay. The biochemical profile was evaluated spectrophotometrically. Detailed RNA-Seq of the data was performed using the transcriptomic method. H&E staining showed well-developed liver structure up to the 160 mg/L boric acid (BA) supplement groups, while BA doses (320 mg/L and 640 mg/L) caused changes in hepatocytes and portal triads. PAS staining showed that glycogen levels were optimal in the 80 mg/L BA dose group, but a reduction in glycogen levels was observed after this group, particularly in the 640 mg/L BA supplement group. Cellular apoptosis showed a biphasic pattern, and the BA dose above 160 mg/L enhanced cell death. In addition, serum analysis showed that doses of 80-160 mg BA were beneficial for ostrich liver. Then, the transcriptome analysis of the 80 mg dose also showed mainly positive effects on the liver. These results demonstrated that chronic BA exposure (320-640 mg) can cause significant histological, apoptotic, and biochemical changes in African ostrich liver, while the adequate dose of supplementation (particularly 80 mg BA) promotes liver growth.

Optimal siting of rainwater harvesting systems for reducing combined sewer overflows at city scale.

The installation of green infrastructure (GI) is an effective approach to manage urban stormwater and combined sewer overflow (CSO) by restoring pre-development conditions in urban areas. Research on simulation-optimization techniques to aid with GI planning decision-making is expanding. However, due to high computational expense, the simulation-optimization methods are often based on design storm events, and it is unclear how much different rainfall scenarios (i.e., design storm events vs. long-term historical rainfall data) impact the optimal siting of GI. The Parallel Pareto Archived Dynamically Dimensioned Search (ParaPADDS) algorithm in a novel simulation-optimization tool OSTRICH-SWMM was used to leverage distributed computing resources. A case study was conducted to optimally site rainwater harvesting cisterns within 897 potential subcatchments throughout the City of Buffalo, New York. Seven design storm events with different return periods and rainfall durations and a one-month historical rainfall time series were considered. The results showed that the optimal solutions of siting cisterns using event-based scenarios, though less computationally expensive, may not perform well under continuous rainfall scenarios, suggesting design rainfall scenarios should be carefully considered for optimizing GI planning. The impact of rainfall scenarios was particularly significant in the middle region of the Pareto front of multi-objective optimization. Utilizing high-performance parallel computing, OSTRICH-SWMM is a promising tool to optimize GI at large spatial and temporal scales.