Hydroxyapatite-chitosan composites derived from sea cucumbers and shrimp shells ameliorate femoral bone defects in an albino rat model.

Background and Aim: A bone defect is defined as a critically sized autologous bone and a bone gap. Bone grafting is one of the most commonly used surgical methods to enhance bone regeneration in orthopedic procedures. A composite of collagen, hydroxyapatite (HA), and chitosan (Ch) is suitable as a bone matrix and stimulates ossification. This study aimed to evaluate the use of natural HA-Ch composites derived from sea cucumbers and shrimp shells and quantify the levels of cytokines, polymorphonuclear neutrophils (PMNs), serum liver enzymes, calcium, phosphate, and procollagen type 1 N-terminal propeptide (PINP) in albino rats with femoral bone defects. Materials and Methods: A total of 48 albino rats with femoral bone defects were divided into 4 groups (n = 12 each): (C-) placebo, (C+) polyethylene glycol, (T1) HA, and (T2) HA-Ch groups. Each group was divided into two subgroups (n = 6 each), with euthanization on 7- and 42-day post-treatment, respectively. Procollagen Type 1 N-terminal propeptide and the cytokines interleukin (IL)-4, IL-6, IL-10, and tumor necrosis factor-alpha were quantified using enzyme-linked immunosorbent assay. Flow cytometry was performed to evaluate PMNs. A clinical chemistry analyzer was used to measure the serum levels of liver enzymes, calcium, and phosphate. Results: There was a significant decrease in the level of IL-6 on 7 days and in the level of IL-10 on 42 days in the HA-Ch group. The level of PMNs also decreased significantly on 7 and 42 days in the HA-Ch group. Regarding serum liver enzymes, alkaline phosphatase (ALP) levels in the HA-Ch group increased significantly on 42 days. Calcium and phosphate levels increased significantly on 7 and 42 days in the HA and HA-Ch groups, and PINP levels increased significantly on 7 and 42 days in the HA-Ch group. Conclusion: The HA-Ch composite derived from sea cucumbers and shrimp shells ameliorated femoral bone defects in albino rats. The HA-Ch composite modulated the levels of IL-6, IL-10, PMNs, ALP, calcium, phosphate, and PINP on 7- and 42-day post-treatment.

Influence of microbiota inoculum as a substitute for antibiotic growth promoter during the initial laying phase on productivity performance, egg quality, and the morphology of reproductive organs in laying hens.

Background and Aim: Antibiotics that increase growth have long been employed as a component of chicken growth. Long-term, unchecked usage may lead to microbial imbalance, resistance, and immune system suppression. Probiotics are a suitable and secure feed additive that may be provided as a solution. The objective of this research was to ascertain the effects of dietary multistrain probiotics (Lactobacillus acidophilus, Bifidobacterium spp., and Lactobacillus plantarum) on the morphology (length and weight) of reproductive organs and productivity performance of laying hens during the early stage of laying. Materials and Methods: One hundred ISA Brown commercial layer chicks of the same body weight (BW) that were 5 days old were divided into five treatments, each with four replicates and four chicks in each duplicate. There were five different dietary interventions: (T1) 100% base feed; (T2) base feed with 2.5 g of antibiotic growth promoter/kg feed; (T3) base feed plus probiotics; (T4) base feed at 1 mL/kg with probiotics; and (T5) base feed with probiotics, 3 mL/kg feed, 5 mL/kg of feed. The parameters observed were performance, internal and exterior egg quality, and the morphology (length and weight) of laying hens' reproductive organs. Results: Probiotic supplementation (L. acidophilus, Bifidobacterium, and L. plantarum) significantly affected the BW, feed intake, egg weight, yolk index, albumin index, Haugh unit, egg height, egg width, and morphology (length and weight) of laying hens' reproductive organs compared to the control group (basic feed). In addition, there was no discernible difference between treatment groups in theeggshell weight and thickness variables across all treatment groups. Conclusion: When laying hens were between 17 and 21 weeks old, during the early laying period, microbiota inoculum supplements (L. acidophilus, Bifidobacterium, and L. plantarum) increased growth, the quality of the internal and external layers' eggs, and the morphology of the laying hens' reproductive organs.

Incidence, risk factors, and therapeutic management of equine colic in Lamongan, Indonesia.

Background and Aim: Colic is among the common health issues in equine health management. Gastrointestinal (GI) disorders are the most frequent causes of colic, but dysfunction of other organs and systems inside the abdominal cavity may also contribute. Therefore, it is crucial to identify risk factors for colic of specific etiologies. This study aimed to examine the incidence, risk factors, and best therapeutic management practices for horses with colic. Materials and Methods: A cohort of 256 horses living in Lamongan, East Java, Indonesia, was randomly recruited based on reports of colic symptoms by owners. Diagnosis and treatment were then conducted with the help of owners. Symptom profiles, risk factors, and therapeutic management strategies were analyzed by Chi-square tests. Results: Of 256 horses enrolled, 217 (84%) were diagnosed with colic, of which 172 (79.3%) were cases of spasmodic colic, 33 (15.2%) of impaction colic, and 12 (5.5%) of intestinal obstruction/displacement. Male sex (χ2 = 16.27; p < 0.001), wheat bran feeding (χ2 = 15.49; p < 0.001), concentrate feed intake >5 kg/day (χ2 = 24.95; p < 0.001), no regular anthelmintic drug treatment (χ2 = 67.24; p < 0.001), GI parasite infection (χ2 = 65.11; p < 0.001), recurrent colic (χ2 = 91.09; p < 0.001), poor body condition score (χ2 = 71.81; p < 0.001), limited daily water access (χ2 = 127.92; p < 0.001), and indications of dental disease (χ2 = 9.03; p < 0.001) were identified as risk factors. The most effective therapies were gastric intubation (χ2 = 153.54; p < 0.001), Vitamin B complex injection (χ2 = 32.09; p < 0.001), fluid therapy (χ2 = 42.59; p < 0.001), and non-steroidal anti-inflammatory drug injection (NSAID). Conclusion: Colic is highly prevalent among horses in Lamongan, East Java, Indonesia. Proper diet, workload management, regular access to clean drinking water, and dental care can reduce colic risk. Recommended therapies include NSAID injection without other analgesics or spasmolytics, fluid therapy, Vitamin B complex, and gastric intubation.

Role of apoptotic inhibitors, viability, and differentiation in low oxygen tension of mesenchymal stem cells cultured in a rat model of ovarian failure.

Background: Stem cell therapy shows applications potential for malnutrition-induced ovarian failure in rat models. However, it is ineffective because of the lack of viability and differentiation of transplanted stem cells, resulting in low adaptation and survival rates. We aimed to determine whether stem cells cultured under low oxygen (O 2) tension improves the adaptability and viability of stem cells, as well as ovarian failure. Methods: After four days of culturing mesenchymal stem cells (MSCs) in 21% oxygen (normoxia) as the T2 group and 1% oxygen (low O 2 or hypoxia) as the T1 group, 200 million bone marrow-derived MSCs per rat were transplanted into female rats with ovarian failure (15 rats per treatment group). A total of 15 fertile and 15 infertile rats were categorized as the C+ and C- groups, respectively. Results: The slight increase in cells expressing HSP70 (C+, T2, T1, and C- groups were 0.5 a±0.53, 1.7 a±0.82, 6.2 b±1.5, and 9.6 c±1.3, respectively), decrease in cells expressing caspase-3 as an apoptotic inhibitor (C+, T2, T1, and C- groups were 0.2 a±0.42, 0.6 a±0.52, 4.8 b±1.03, and 7.3 c±1.42, respectively), and increase in cells expressing VEGF-1 (C+, T2, T1, and C- groups were 10.8 c±1.55, 8.7 b±0.48, 0.4 a±0.52, and 0.2 a±0.42, respectively) and GDF-9 (C+, T2, T1, and C- groups were 5.8 c±1.47, 4.6 b±0.97, 0.5 a±0.53, and 0.3 a±0.48, respectively) were used as markers for viability and differentiation in ovarian tissue, indicating that MSCs cultured under low O 2 tension were more effective than those cultured under normoxic conditions as a treatment for female rats with ovarian failure. Furthermore, infertile female rats treated with MSCs cultivated under low O 2 tension had an enhanced ovarian tissue shape, as indicated by the increasing Graafian follicle count (C+, T2, T1, and C- groups were 8.9 c±0.74, 4.5 b±0.71, 0.5 a±0.53, and 0.4 a±0.52, respectively). Conclusions: MSCs cultured under low O 2 tension are an effective treatment for malnourished rats with ovarian failure.

Aerolysin gene characterization and antimicrobial resistance profile of Aeromonas hydrophila isolated from milkfish (Chanos chanos) in Gresik, Indonesia.

Background and Aim: Motile Aeromonas septicemia is a crucial disease in freshwater fish. Aeromonas hydrophila is a disease agent associated with sporadic fish mortality, food safety, and public health. This study aimed to estimate the prevalence and the presence of the aerolysin gene and antimicrobial resistance profile of A. hydrophila isolated from milkfish in Gresik, Indonesia. Materials and Methods: A total of 153 milkfish gill samples were collected from 16 locations in Gresik and then cultured and identified using biochemical tests. The aerolysin gene was investigated using a polymerase chain reaction, and antimicrobial resistance profiles of the recovered isolates were investigated. Results: Of the 153 examined samples, 35 (22.9%) were confirmed positive for A. hydrophila and 22 (62.9%) presented the aerolysin gene. The recovered isolates were resistant to the following antibiotics: Amoxicillin (62.9%), tetracycline (60%), streptomycin (54.3%), cefotaxime (51.4%), gentamycin (31.4%), kanamycin (28.6%), erythromycin (25.7%), chloramphenicol (20%), and trimethoprim (14.3%). Meanwhile, only ciprofloxacin, nalidixic acid, and imipenem were indicated as susceptible. Conclusion: The presence of the aerolysin gene is vital in determining the virulence of A. hydrophila. The study results indicated a high aerolysin gene prevalence. In addition, this study emphasized antibiotic use monitoring, food safety improvement, and negative impact reduction on human health and the environment.

Effectiveness of forest honey ( Apis dorsata) as therapy for ovarian failure causing malnutrition.

Background: Malnutrition is the imbalance between intake and nutritional needs, resulting in a decrease in body weight, composition, and physical function. Malnutrition causes infertility due to intestinal and liver degeneration,which may progress to testicular and ovarian degeneration. Methods: An infertile female rat model with a degenerative ovary was induced with malnutrition through a 5-day food fasting but still had drinking water. The administration of (T1) 30% (v/v) and (T2) 50% (v/v) forest honey ( Apis dorsata) were performed for ten consecutive days, whereas the (T+) group was fasted and not administered forest honey and the (T-) group has not fasted and not administered forest honey. Superoxide dismutase, malondialdehyde, IL-13 and TNF-α cytokine expressions, and ovarian tissue regeneration were analyzed. Results: Superoxide dismutase was significantly different ( p<0.05) in T1 (65.24±7.53), T2 (74.16±12.3), and T- (65.09±6.56) compared with T+ (41.76±8.51). Malondialdehyde was significantly different ( p<0.05) in T1 (9.71±1.53), T2 (9.23±0.96), and T- (9.83±1.46) compared with T+ (15.28±1.61). Anti-inflammatory cytokine (IL-13) expression was significantly different ( p<0.05) in T1 (5.30±2.31), T2 (9.80±2.53), and T- (0.30±0.48) compared with T+ (2.70±1.57). Pro-inflammatory cytokine (TNF-α) expression was significantly different ( p<0.05) in T1 (4.40±3.02), T2 (2.50±1.65), and T- (0.30±0.48) compared with T+ (9.50±1.78). Ovarian tissue regeneration was significantly different ( p<0.05) in T- (8.6±0.69) and T2 (5.10±0.99) compared with T1 (0.7±0.95) and T+ (0.3±0.67). Conclusion: The 10-day administration of 50% (v/v) forest honey can be an effective therapy for ovarian failure that caused malnutrition in the female rat model.

An investigation of syrinx morphometry and sound frequency association in lovebirds ( Agapornis fischeri) chirps.

Background: In the issue of biodiversity, the domestication of birds as pets and trade animals requires special attention as a conservation effort. Lovebirds ( Agapornis fischeri) are popular birds worldwide, due to their varied ornamentation and melodic chirping sound. Syrinx structure is suspected to be the main source of sound production during the chirping period. This study aimed to investigate syrinx morphometry and its correlation with sound frequency produced in lovebirds. Methods: A total of 24 lovebirds of different ages and gender were investigated. Polymerase chain reaction method was performed to determine lovebird gender, meanwhile bird age was identified based on post-hatch recordings at the breeding farm. Thus, we enrolled male (n=12) and female (n=12) lovebirds aged 2 (n=4), 3 (n=4), and 4 (n=4) months in the investigation group, respectively. Fast Fourier Transform (FFT) was performed to evaluate sound frequency during chirping period. Then, syrinx morphometry was identified using a topographic approach and methylene blue staining. Each variable was evaluated with Image J software and vernier caliper. Results: Based on a topographical approach, we reported the general cartilage structure of the tracheosyringeal, bronchosyringeal, paired protrusions, tracheolateral muscles, sternotracheal muscles, and syringeal muscles in lovebird syrinx. In particular, the tympaniform membranes lateral lead a crucial role in modulating the frequency of male lovebirds more significantly (p=0,009) compared to female. On the other hand, the tympaniform membranes lateral dexter (p=0,02) and sinister (p=0,05) in females showed wider compared to male. We also reported a negative correlation between sound frequency compared to tympaniform membranes lateral dexter (y = -913,56x + 6770,8) and sinister (y = -706,16x + 5736). Conclusions: It can be concluded that the tympaniform membranes lateral produced the lovebirds' primary sound. The sound frequency of male lovebirds was higher compared to female, however negatively correlated with the area of tympaniform membranes lateral.

Effects of dietary supplementation with breadfruit leaf powder on growth performance, meat quality, and antioxidative activity in Japanese quail.

BACKGROUND AND AIM: In an era of increasing concerns about food availability globally, poultry meat is being increasingly consumed rather than red meat given its quality in terms of pH, color, and tenderness, conferring consumer satisfaction. The choice of feed is a crucial factor in poultry production. This study investigated the effect of dietary supplementation with breadfruit leaf powder on growth performance, meat quality, and antioxidative activity in Japanese quail. MATERIALS AND METHODS: A total of 120 day-old quail were used in this study and assigned equally into four treatment groups: Group C fed a basal diet and three treatment groups fed a basal diet supplemented with 2.5% (T1), 5% (T2), or 10% (T3) breadfruit leaf powder. The concentrations of breadfruit leaf powder were 2.5, 5, and 10 g/kg in the basal diet. Quail body weight and feed intake (FI) were evaluated at 1, 21, and 35 days of age at 7 a.m. Pectoral muscle was collected to determine pH, meat color, drip loss, cooking loss, water-holding capacity (WHC), tenderness, and antioxidant levels. All variables were analyzed statistically using ANOVA followed by Duncan's post hoc test (significance set at p<0.05). RESULTS: T3 showed increased body weight gain of quails at1-21 and 21-35 days (p<0.05). Feeding in the T3 group improved the feed conversion ratio compared with those in the C and T1 groups at the starter phase (p<0.05). Dietary treatment did not affect FI (p>0.05). In the present study, meat redness and WHC were improved in the T3 group (p<0.05). Meanwhile, drip loss, cooking loss, and meat tenderness were improved in the T2 group (p<0.05). The pH45 min, pH24 h, lightness, and yellowness were not influenced by the treatments (p>0.05). The antioxidative activities of superoxide dismutase and malondialdehyde decreased in the T3 group (p<0.05), while no significant difference in glutathione peroxidase level (p>0.05) was identified. CONCLUSION: Ten grams/kilogram of breadfruit leaf powder, as administered in the T3 group, can be applied as a dietary supplement for Japanese quail to improve growth performance, meat quality, and antioxidative activity during the starter and grower periods.

Risk factors, hematological and biochemical profile associated with colic in Delman horses in Gresik, Indonesia.

Background: Horses are herd animals that have been domesticated in the last century. In several countries, an overview of risk factors and clinical evaluation in horses with colic has not been well-described. This study aimed to evaluate risk factors and hematological profiles in horses associated with colic in Gresik, East Java, Indonesia. Methods: A cross-sectional study was performed during April - October 2019. A total of 115 horses were diagnosed based on physical examination, clinical symptoms, and rectal examination. A questionnaire was asked to the horse-owners to analyze the risk factors while the clinical examination was performed and blood samples were collected for pre-treatment and 14 days post-treatment. Hematological profile was evaluated from a whole blood sample. Serum cortisol, plasma epinephrine, and norepinephrine concentrations were also evaluated after separating the aliquots.  Results: Of the 115 horses, 96 were diagnosed with colic. The horses with colic showed a significant association between cases with gender (p<0.021), breed (p<0.000), wheat bran feeding (p<0.015), concentrate feeding (p<0.003), anthelmintics administration (p<0.000), gastrointestinal parasites (p<0.000), dental diseases (p<0.024), previous exposure to colic (p<0.000), body condition score (p<0.000), and access to water per day (p<0.000). Based on whole blood and serum evaluation, there were ameliorated significantly on the hematological profile (p<0.01), serum cortisol (p<0.05), and plasma epinephrine (p<0.01) at 14 days post-treatment. Conclusion: This study has identified factors associated with colic in Delman horses. The study provides crucial information to investigate cases of colic and to contribute the development of healthcare strategies during treatment and clinical evaluation.