Pre‐soaking of the feed pellets: a trick for successful feed utilization in juvenile green turtles (Chelonia mydas Linnaeus, 1758)

Pre‐soaking of the feed pellets in water can improve feed utilization in juvenile green turtles (Chelonia mydas Linnaeus, 1758), but the pre‐soaking has not previously been optimized. This study aimed to optimize the water amount used for pre‐soaking the pellets. The experiments followed a completely randomized design with three replications of each dietary treatment group. Initially 10‐day‐old green turtles (20–22 g body weight) were treated in an indoor aquaculture system for 3 months. The dietary treatment pellets were pre‐soaked with 0.3, 0.5 or 0.7 (v/w) relative amounts of water that are here termed soaking ratios. At the end of experiment, there were no significant differences in survival (96% on average) and growth (average body weight 75.34 g and specific growth rate 2%/day, on average) of turtles in three dietary treatments (p > 0.05). Feed utilization was the best in turtles fed with 0.7 pre‐soaked ratio, as indicated by significant reductions (p < 0.05) in the feeding rate (7.44% body weight/day) and the feed conversion ratio (1.12 g feed/g gain). Digestion was also improved by the induction of faecal digestive enzymes as well as the faecal thermal properties. The rapid growth did not negatively affect the general haematological parameters of reared turtles. These findings indicate that the pre‐soaking of feed pellets at the optimal soaking ratio (1:0.7 w/v of pellet to water) can contribute through improved feed utilization of green turtles.     

Immunohistochemical study of Ki‐67 protein,androgen receptor,and estrogen receptor beta in testicular tissues of male pigs immunocastrated with different times of GnRH vaccination

This study aimed to investigate the immunoexpression of Ki‐67 protein, androgen receptor (AR), and estrogen receptor beta (ERβ) in testicular tissues of male pigs immunocastrated using GnRH vaccine (Improvac?, Zoetis Co., Ltd., Thailand) with different times. Totally, 30 male pigs were classified by castration protocol into three groups: T1 (n = 10) consisted of pigs immunocastrated at 14 and 18 weeks of age, T2 (n = 10) included pigs immunocastrated at 9 and 19 weeks of age, and C (n = 10) contained intact pigs. The results revealed that testicular length of pigs in C was longer than that of both T1 (8.1 ± 0.76 vs 6.5 ± 0.5 cm, p < 0.001) and T2 (8.1 ± 0.76 vs 6.9 ± 1.0, p = 0.007). Spearman correlation coefficients showed negative correlation between testicular length and H‐score of AR (r = ?0.38, p = 0.037), as well as positive correlation between testicular length and Ki‐67 index (r = 0.602, p < 0.001). Generally, mean Ki‐67 index and mean H‐scores of AR and ERβ of pigs in T1 were not different from those in T2 (p > 0.05). However, mean Ki‐67 index and mean AR H‐scores of T1 and T2 were significantly different from C group (p < 0.05). In summary, the immunocastration significantly affected testicular length, including expressions of Ki‐67, AR, and ERβ in pig testes. Moreover, the duration between two shots of GnRH vaccine could be extended from 4 to 10 weeks without difference in Ki‐67 protein, AR, and ERβ immunoexpressions.     

Morphometric analysis of cervical vertebrae in some marine and land mammals

Bones or skeletal remains can be used to answer a number of questions related to species, sex, age or cause of death. However, studies involving vertebrae have been limited as most were performed on skulls or long bones. Here, we have stated the hypothesis that the morphometry of cervical vertebrae can be used for species identification and body size estimation among marine and land mammals. The cervical vertebrae from eight and 14 species of marine and land mammals were used to collect morphometric data. Cluster dendrogram, principal component analysis, discriminant analysis and linear regression were used to analyse the data. The results indicate that, based on an index of C4 to C7, there were 13 out of 22 species for which identity could be correctly predicted in 100% of the cases. The correlations between cervical vertebrae parameters (height, width and length of centrum) in marine (average R^2 = 0.87, p < .01) and land (average R^2 = 0.51, p < .01) mammals were observed. These results indicate that vertebral morphometrics could be used for species prediction and verification of body weight in both marine and land mammals.     

Expressions of cytochrome P450 aromatase and anti-Müllerian hormone in testes of fattening pigs by the timing of the first vaccination for immunocastration

In practice, two injections of gonadotropin-releasing hormone (GnRH) vaccine are recommended for pig immunocastration for effective outcomes. The present study aimed to investigate the expressions of cytochrome P450 aromatase (P450_arom) and anti-Müllerian hormone (AMH) in testes, testicular length and testicular histomorphometry of the fattening pigs receiving the first injection of GnRH vaccine 6 weeks earlier than the standard protocol. Based on vaccination protocol, 24 pigs were equally divided into three groups: T1 was vaccinated at 15 and 19 weeks of age, T2 received vaccine at 9 and 19 weeks of age and C remained intact. P450_arom and AMH expressions were analysed using immunohistochemistry and Western blot. The results revealed that testicular length was highest in C pigs, but not different between T1 and T2 groups (6.5 ± 0.2 versus 6.9 ± 0.3 cm, p = .538). Histomorphometry demonstrated that the height of spermatogenic epithelia, the diameter of seminiferous tubules and the number of seminiferous tubules between T1 and T2 groups were not different (p > .05). For P450_arom, immunohistochemistry revealed that H-score of C group was significantly higher than that of both T1 and T2 groups. Western blot analysis showed that C group possessed the densest protein band. Moreover, H-score between T1 and T2 groups was not significantly different. Protein band intensity between both groups was not apparently different. As for AMH, C pigs had significantly lower H-score than both T1 and T2 pigs. Furthermore, T2 pigs possessed significantly higher H-score than T1 pigs. Western blot analysis showed that the most intense protein band was found in T2 group. In summary, GnRH vaccine affected testicular development and functions. The first injection could be performed either at 9 or 15 weeks of age since both protocols contributed to comparable results in aspect of testicular length, histomorphometry and expressions of P450_arom and AMH.     

Early divergence and differential population histories of the Indo-Pacific humpback dolphin in the Pacific and Indian Oceans

The currently recognized Indo-Pacific humpback dolphin occurs in estuaries and surrounding shallow waters from the South China Sea to the Asian coast of the Indian Ocean. However, a recent study suggested that the humpback dolphin from the Bay of Bengal may represent a distinct phylogenetic species. In this study, we sequenced 915-bp mtDNA segments from five geographic populations in both Chinese and Thai waters; together with previously published sequences, these data revealed that the ancestral Indo-Pacific humpback dolphin might have split during the transition from the Oligocene to Miocene (23.45 Mya, 95% HPD: 16.65–26.55 Mya), and then dispersed along the Pacific and Indian Ocean coasts of Asia. Genetic differentiation was detected between most of the examined populations, except for only a few pairwise populations in the northern South China Sea. Genetic differentiation/distance between the humpback dolphins from the northern and southern South China Sea met the sub-species threshold value proposed for marine mammals, whereas that between the humpback dolphins in the Pacific and the Indian Ocean was above the species threshold. Bayesian inference of historic gene flow indicated low but constant northward gene flow along the Indian Ocean coast; however, there was a recent abrupt increase in gene flow in the Pacific region, likely due to the shortening coastline at the low stand of sea level. Our results revealed that the current taxonomic classification of Indo-Pacific humpback dolphins may not reflect their phylogeography.     

Optimal feeding frequency of captive head‐started green turtles (Chelonia mydas)

Optimal feeding frequency was investigated to improve head‐started propagation programme of juvenile green turtles (Chelonia mydas). The 15‐day‐old turtles (25–26 g body weight) were fed for ad libitum intake at one (1MD), two (2MD), three (3MD) or four (4MD) meals daily over a 3‐month trial. Responses in growth, feed utilization, faecal characteristics, haematological parameters and carapace elemental composition were used to compare treatment effects. At the end of the feeding trial, no treatment had induced mortality. Growth performance in terms of weight gain and specific growth rate was similar in turtles fed 2MD, 3MD or 4MD (p > 0.05), but 1MD differed from these (p < 0.05), and feeding at excess frequency (3MD and 4MD) increased the within‐group size variation. Turtles fed 2MD had significantly lower feed intake than in 3MD and 4MD groups, but the feed conversion ratios were similar. Faecal digestive enzyme analysis indicated higher catabolism of lipid and protein in the deprivation group (1MD), when compared with turtles fed at least twice daily. The feeding frequency did not affect the specific activities of carbohydrate‐digesting enzymes. The results on enzymes activities were corroborated by the transition enthalpy characteristics of faeces, indicating nutrients remaining after digestion. The 2MD treatment also improved the haematological characteristics and the carapace quality, relative to low or excess feeding. Overall, the findings indicate that feeding juvenile green turtles twice a day is the preferred option in their head‐started propagation. This promotes growth, reduces feed consumption, and improves health and carapace quality.