Suitable methods for cryopreservation of semen from Atlantic halibut, Hippoglossus hippoglossus L.

Decrease in the quality and quantity of Atlantic halibut, Hippoglossus hippoglossus L., semen towards the end of the reproductive season hampers production of good-quality embryos. Therefore, cryopreservation of spermatozoa is a method showing potential to facilitate controlled reproduction in Atlantic halibut. The present study aimed at establishing the appropriate cryopreservation procedure. We tested 20 extenders composed of four various diluents and five cryoprotectants (DMSO, DMA, methanol, propylene glycol, and glycerol) to determine the best extender. Then, we examined cryopreservation quality using various methods of loading and various volumes of cryopreserved samples. In most of the tested variants, sperm diluted with an extender showed high motility after 24-h incubation despite the high osmotic pressure of the extender. Modified turbot extender (MTE) was the best of the tested diluents, securing the highest post-thaw motility (P < 0.05), and DMSO, DMA, and methanol were the best cryoprotectants (P < 0.05). There was no significant effect of 15-min equilibration of semen in MTE-based extenders prior to freezing on post-thaw motility (P > 0.05). MTE-based extender was chosen as the most suitable. Semen cryopreserved in straws, Eppendorfs or Ziploc bags in volumes ranging from 0.25 to 20 ml showed similar high fertilization ability. Survival of larvae produced with the cryopreserved sperm did not differ from controls produced with freshly collected sperm. Motility 3 h after thawing was high but depended on the type of cryoprotectant and the volume of cryopreserved sperm (P < 0.05). The developed cryopreservation procedure has been applied at our Atlantic halibut breeding station for seed production.     

Hematology and serum biochemistry of captive Sunda pangolin (Manis javanica) in Wildlife Reserves Singapore

The Sunda pangolin (Manis javanica) faces threat of extinction due to illegal trafficking of its scales for the purpose of traditional medicine in the Asian region. Ex-situ captive breeding and reintroduction programs have been identified to be a key effort in the conservation of the species. The establishment of blood parameters for captive Sunda pangolins are vital to assess the health in these animals during health assessments. The objective of this study is to establish blood parameters for captive Sunda pangolins and compare the blood parameters with the established blood reference ranges for rescued wild Sunda pangolins in Singapore. Blood parameters for hematology and serum biochemistry were established from 13 clinically normal captive Sunda pangolins. Male captive Sunda pangolins were found to have significantly (P<0.05) higher potassium compared to the female captive Sunda pangolins. Captive Sunda pangolins were found to have significantly (P<0.05) lower white blood cell count (WBC), neutrophil counts, alanine phosphatase (ALP) and phosphorus and significantly (P<0.05) higher PCV, TP, globulin and blood urea nitrogen (BUN) when compared with rescued wild Sunda pangolins from the previous study.     

Severe hydrocephalus in a raccoon dog (Nyctereutes procyonoides)

Hydrocephalus is one of the most common central nervous system malformations in domestic dogs, yet they are poorly documented and studied in wild carnivoran mammals. A pup of raccoon dog (Nyctereutes procyonoides) was rescued and brought to Wildlife Center. The pup showed generalized ataxia, a domed skull, and an open bregmatic fontanelle. Ultrasound and MRI showed severe enlargement of the lateral ventricle with the loss of septum pellucidum resulting in a single large ventricle and cervical syringohydromyelia. Although treatment was attempted, the animal was euthanized due to poor prognosis. At necropsy, macroscopic findings were identical to the diagnostic imaging, where marked enlargement of the calvarium, and attenuated gyri and sulci were observed. Finally, hydrocephalus was confirmed. Here, we describe a case of hydrocephalus in a raccoon dog (Nyctereutes procyonoides).     

Cloning of the short-tailed Gyeongju Donggyeong dog via SCNT: conserving phenotypic inheritance

Somatic cell nuclear transfer is a useful tool to maintain genetic information of animals. The Gyeongju Donggyeong dog is a breed registered as natural monument in Korea. The unique feature of the Donggyeong dog is its tail, as the Donggyeong dog can be classified as either short tailed or tailless. The aim of this study was to preserve the Donggyeong dog’s unique feature by cloning. Fibroblasts were obtained from a short-tailed Donggyeong dog. In vivo matured oocytes were enucleated, microinjected with a donor cell and fused electrically. Reconstructed embryos were transferred to six recipient dogs. One surrogate became pregnant, and one short-tailed Donggyeong dog was delivered. This study demonstrated that the phenotype of the Donggyeong dog could be conserved by somatic cell nuclear transfer.     

日本茶树种植导致的环境问题及施用氨晴钙的控制效果

A field experiment, involving lime N （calcium cyanamide, CaCN2） fertilization as a control measure, was conducted to study environmental problems induced by long-term heavy N application in Japanese tea fields. Long-term tea cultivation caused serious soil acidification. Seventy-seven percent of the 70 tea fields investigated had soil pH values below 4.0, and 9% below 3.0, with the lowest value of 2.7. Moreover, excess N application in tea fields put a threat to plant growth, induced serious nitrate contamination to local water, and caused high nitrous oxide loss. Compared with the conventional high N application treatment （1 100 kg N ha^-1） without lime N, the low N application （400 kg N ha^-1） with calcium cyanamide effectively stopped soil acidification as well as achieved the same or slightly higher levels in tea yield and in total N and amino acid contents of tea shoots. The application of calcium cyanamide could be a suitable fertilization for the prevention of environmental problems in tea cultivation.     

Brain Abscess in a 4-Month-Old Filly: A Case Report

Although recognized as a possible complication to sepsis, there are few published reports on brain abscesses in foals. This case report describes a 4-month-old Icelandic filly that, as a neonate, had a history of pyrexia and lameness, which resolved with antibiotic treatment. Approximately 3 months later, the foal developed neurological signs which were initially attributed to trauma; she was referred for further investigation. The clinical signs were consistent with a lesion of the right forebrain. Computed tomography examination revealed a mass, approximately 6 cm in diameter, located in the right cerebral hemisphere craniolateral to the right lateral ventricle. Because of the poor prognosis, the foal was killed, and a postmortem was performed. The findings confirmed the presence of an abscess of the right cerebral hemisphere, leading to severe atrophy of the cortex and dilation of both lateral ventricles. The left hemisphere was moderate-to-severe atrophic and had displaced to the left. Streptococcus equi subspecies zooepidemicus was isolated in pure culture from the abscess. The clinical features of the foal were compared with the pathological findings.